Oncology Intelligence

Colon Cancer
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11:00 PM|Farchoukh, Lama; Kuan, Shih-Fan; Dudley, Beth; Brand, Randall; Nikiforova, Marina; Pai, Reetesh K.|The American Journal of Surgical Pathology - Current Issue|Labels: BRAF, RAS, CRC
imageBetween 10% and 15% of colorectal carcinomas demonstrate sporadic DNA mismatch-repair protein deficiency as a result of MLH1 promoter methylation and are thought to arise from sessile serrated adenomas, termed the serrated neoplasia pathway. Although the presence of the BRAF V600E mutation is indicative of a sporadic cancer, up to 30% to 50% of colorectal carcinomas with MLH1 promoter hypermethylation will lack a BRAF mutation. We report the clinicopathologic and molecular features of MLH1-deficient colorectal carcinoma with wild-type BRAF and MLH1 promoter hypermethylation (referred to as MLH1-hypermethylated BRAF wild-type colorectal carcinoma, n=36) in comparison with MLH1-deficient BRAF-mutated colorectal carcinoma (n=113) and Lynch syndrome–associated colorectal carcinoma (n=36). KRAS mutations were identified in 31% of MLH1-hypermethylated BRAF wild-type colorectal carcinomas compared with 0% of MLH1-deficient BRAF-mutated colorectal carcinomas and 37% of Lynch syndrome–associated colorectal carcinomas. When a precursor polyp was identified, MLH1-hypermethylated BRAF wild-type colorectal carcinomas arose from precursor polyps resembling conventional tubular/tubulovillous adenomas in contrast to MLH1-deficient BRAF-mutated colorectal carcinomas, which arose from precursor sessile serrated adenomas (P<0.001). Both MLH1-hypermethylated BRAF wild-type colorectal carcinoma and MLH1-deficient BRAF-mutated colorectal carcinoma had a predilection for the right colon compared with Lynch syndrome–associated colorectal carcinoma (86% vs. 92% vs. 49%, P<0.001). There was no significant difference in mucinous differentiation, tumor-infiltrating lymphocytes, Crohn-like reaction, and medullary differentiation between the 3 tumor groups. Using Kaplan-Meier survival functions, there was no significant difference in disease-specific survival between the 3 patient groups (P>0.05). In conclusion, our results indicate that MLH1-hypermethylated BRAF wild-type colorectal carcinomas can harbor KRAS mutations and arise from precursor polyps resembling conventional tubular/tubulovillous adenomas.
Thursday, September 22, 2016 9:16 AM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: RAS, CRC, clinical trial
European Journal of Surgical Oncology, Volume 42, Issue 10, 2016 Oct | Köhne, C.-H.; Poston, G.; Folprecht, G.;...
Introduction Improvements in surgical intervention have significantly contributed to the enhanced survival of patients with metastatic colorectal cancer (mCRC) that has been observed over the past 15 years. The prognosis for patients presenting wi...
Thursday, September 22, 2016 9:16 AM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 10, 2016 Oct | Agus, M.; Lo Mastro, M.; Cabras, F.;...
Neoadjuvant chemoradiation (RCT) is standard care for locally advanced rectal cancer. For many patients RCT results in a significant tumor regression, while others show no response or even disease progression and develop metastatic disease. Unfort...
Metastatic colorectal cancer (mCRC) combined with hyperbilirubinemia is typically considered a contraindication to irinotecan-based therapy, a proven first-line treatment of mCRC. Herein, we present 6 consecutive patients with mCRC combined with hyperbilirubinemia who underwent UGT1A1 genotyping before receiving FOLFIRI plus bevacizumab. Dose escalation of irinotecan was performed according to the results of UGT1A1 genotyping in all patients. Improvement in the serum total bilirubin level to a normal range was noted in all 6 patients. Disease control was 100%. The median progression-free survival was 7.5 months and the median overall survival was 8.5 months. FOLFIRI plus bevacizumab as a first-line chemotherapy may achieve effective disease control and be safe in patients with mCRC and hyperbilirubinemia based on UGT1A1 genotyping. More prospective clinical studies are necessary to evaluate the clinical benefits and safety of this treatment approach.
Chemotherapy 2017;62:80-84
Wednesday, September 21, 2016 8:09 AM|Elsevier|JournalTOCs API - Biochemical and Biophysical Research Communications (50 articles)|Labels: CRC

MicroRNA-215 suppresses cell proliferation, migration and invasion of colon cancer by repressing Yin-Yang 1

Biochemical and Biophysical Research Communications, Vol. , No. (2016) pp. -
Publication date: Available online 20 September 2016 Source:Biochemical and Biophysical Research Communications Author(s): Zehong Chen, Siqi Han, Wensheng Huang, Jialin Wu, Yuyi Liu, Shirong Cai, Yulong He, Suijing Wu, Wu Song Colorectal cancer is one of the most common malignant tumors worldwide with rising incidence. MicroRNAs are small non-coding RNAs that implicate in multiple physiological or pathological processes. The aberrant expression of miRNA-215 (miR-215) has been illustrated in various types of cancers. However, the expression of miR-215 in human colon cancer and the biological roles of it remain largely unknown. We conducted this study to explore the expression and the function of miR-215 in human colon cancer. The results showed that miR-215 was remarkably downregulated in colon cancer tissues and cell lines. Overexpression of miR-215 by miR-215 mimic significantly inhibited colon cancer cell proliferation, migration and invasion while knockdown of miR-215 by miR-215 inhibitor exerted reverse effects. Furthermore, we newly identified Yin-Yang 1(YY1) as a direct target of miR-215 which could rescue the effects of miR-215 on colon cancer cells. In summary, our investigation revealed that miR-215 was downregulated in colon cancer and it suppressed colon cancer cell proliferation, migration and invasion by directly targeting YY1.

Tuesday, September 20, 2016 12:14 PM|Salvucci, M., Würstle, M. L., Morgan, C., Curry, S., Cremona, M., Lindner, A. U., Bacon, O., Resler, A. J., Murphy, A. C., O'Byrne, R., Flanagan, L., Dasgupta, S., Rice, N., Pilati, C., Zink, E., Schöller, L. M., Toomey, S., Lawler, M., Johnston, P. G., Wilson, R. H., Camilleri-Broët, S., Salto-Tellez, M., McNamara, D. A., Kay, E. W., Laurent-Puig, P., Van Schaeybroeck, S., Hennessy, B. T., Longley, D. B., Rehm, M., Prehn, J. H.|Clinical Cancer Research Online First Articles|Labels: CRC

Purpose: Apoptosis is essential for chemotherapy responses. In this discovery and validation study, we evaluated the suitability of a mathematical model of apoptosis execution (APOPTO-CELL) as a stand-alone signature and as a constituent of further refined prognostic stratification tools. Experimental Design: Apoptosis competency of primary tumor samples from n=120 stage III colorectal cancer patients was calculated by APOPTO-CELL from measured protein concentrations of Procaspase-3, Procaspase-9, SMAC and XIAP. An enriched APOPTO-CELL signature (APOPTO-CELL-PC3) was synthesized to capture apoptosome-independent effects of Caspase-3. Furthermore, a machine learning Random Forest approach was applied to APOPTO-CELL-PC3 and available molecular and clinicopathological data to identify a further enhanced signature. Association of the signature with prognosis was evaluated in an independent colon adenocarcinoma cohort (TCGA COAD, n=136). Results: We identified three prognostic biomarkers (p=0.04, p=0.006 and p=0.0004 for APOPTO-CELL, APOPTO-CELL-PC3 and Random Forest signatures, respectively) with increasing stratification accuracy for stage III colorectal cancer patients. The APOPTO-CELL-PC3 signature ranked highest among all features. The prognostic value of the signatures was independently validated in stage III TCGA COAD patients (p=0.01, p=0.04 and p=0.02 for APOPTO-CELL, APOPTO-CELL-PC3 and Random Forest signatures, respectively). The signatures provided further stratification for patients of CMS1-3 molecular subtype. Conclusions: The integration of a systems-biology-based biomarker for apoptosis competency with machine learning approaches is an appealing and innovative strategy towards refined patient stratification. The prognostic value of apoptosis competency is independent of other available clinicopathological and molecular factors, with tangible potential of being introduced in the clinical management of stage III colorectal patients.

Contributors : Bernard Omolo ; Mingli Yang ; Fang Y Lo ; Michael J Schell ; Sharon Austin ; Kellie Howard ; Anup Madan ; Timothy J Yeatman
Series Type : Expression profiling by high throughput sequencing
Organism : Homo sapiens

Background: The KRAS gene is mutated in about 40% of colorectal cancer (CRC) cases, which has been clinically validated as a predictive mutational marker of intrinsic resistatnce to anti-EGFR inhibitor (EGFRi) therapy. Since nearly 60% of patients with a wild type KRAS fail to respond to EGFRi treatment, there is a need to develop more reliable molecular signatures to better predict response. Here we address the challenge of adapting a gene expression signature predictive of RAS pathway activation, created using fresh frozen (FF) tissues, for use with more widely available formalin fixed paraffin-embedded (FFPE) tissues.
Methods: In this study, we evaluated the translation of an 18-gene RAS pathway signature score from FF to FFPE in 54 CRC cases, using a head-to-head comparison of five technology platforms. FFPE-based technologies included the Affymetrix GeneChip (Affy), NanoString nCounter(NanoS), Illumina whole genome RNASeq (RNA-Acc), Illumina targeted RNASeq(t-RNA), and Illumina stranded Total RNA-rRNA-depletion (rRNA).
Results: Using Affy_FF as the "gold" standard, initial analysis of the 18-gene RAS scores on all 54 samples shows varying pairwise Spearman correlations, with (1) Affy_FFPE(r=0.233, p=0.090); (2) NanoS_FFPE(r=0.608, p<0.0001); (3) RNA-Acc_FFPE(r=0.175, p=0.21); (4) t-RNA_FFPE (r=-0.237, p=0.085); and (5) t-RNA (r=-0.012, p=0.93). These results suggest that only NanoString has successful FF to FFPE translation. The subsequent removal of identified "problematic" samples (n=15) and gene (n=2) further improves the correlations of Affy_FF with three of the five technologies: Affy_FFPE (r=0.672, p<0.0001); NanoS_FFPE (r=0.738, p<0.0001); and RNA-Acc_FFPE (r=0.483, p=0.002).
Conclusions: Of the five technology platforms tested, NanoString technology provides a more faithful translation of the RAS pathway gene expression signature from FF to FFPE than the Affymetrix GeneChip and multiple RNASeq technologies. Moreover, NanoString was the most forgiving technology in the analysis of samples with presumably poor RNA quality. Using this approach, the RAS signature score may now be reasonably applied to FFPE clinical samples.

Friday, September 16, 2016 4:27 PM|Stephanie Roessler, Guoling Lin, Marshonna Forgues, Anuradha Budhu, Shelley Hoover, R. Mark Simpson, Xiaolin Wu, Ping He, Lun-Xiu Qin, Zhao-You Tang, Qing-Hai Ye, Xin Wei Wang|International Journal of Biological Sciences|Labels: CRC

Metastasis is the main cause of cancer mortality but its process remains poorly understood and thus hampers more effective treatment and improved cancer prognosis. To search for metastasis driver genes responsible for tumor spread, we integrated genomic and transcriptomic profiles of 61 matched primary tumors and distant metastases of liver or colorectal carcinoma isolated by laser-capture microdissection and assayed by array-based technologies. We found that primary tumor lesions and their matched distant metastases were largely similar at the genomic and transcriptomic levels, but substantial differences could be found between primary tumors with or without accompanying metastases. Interestingly, metastasis genes were principally tumor type and organ site-specific. Despite distinct pathway enrichment, different metastasis gene sets shared common prognostic capacity and were predictive of hepatocellular carcinoma survival in an independent cohort. Thus, the metastatic propensity is inherent to the primary tumor and the lack of general metastasis genes necessitates the development of specific treatment modalities.

Friday, September 16, 2016 4:27 PM|Huai-Qiang Ju, Yun-Xin Lu, Dong-Liang Chen, Tian Tian, Hai-Yu Mo, Xiao-Li Wei, Jian-Wei Liao, Feng Wang, Zhao-Lei Zeng, Helene Pelicano, Mitzi Aguilar, Wei-Hua Jia, Rui-Hua Xu|Theranostics|Labels: ROS, CRC

Colorectal cancer (CRC) is a common neoplastic disease and a frequent cause of death. Drug resistance is a major challenge to CRC treatment and stem-like side-population (SP) cells may play a key role in this resistance. Although it has been recognized that cancer stem cells may be affected by redox status, the underlying mechanisms for this effect and the roles of celllular redox adaptation and antioxidant capacity in CRC remain elusive. Our study shows that CRC SP cells are highly dependent on cellular GSH to maintain ROS levels below those of non-SP cells. Exposing CRC cells to H2O2 produced a significant decrease in the percentage of SP cells, which was rescued by adding N-acetylcysteine. Mechanistically, CD44v interacts with and stabilizes xCT and thereby promotes the uptake of cysteine for GSH synthesis and stimulates SP cell enrichment. Additionally, miR-1297 levels were inversely correlated with the expression of xCT; thus, reduced miR-1297 contributes to SP cell enrichment in CRC tumors, which results in tumor aggressiveness and poor clinical outcomes. Importantly, redox modification by PEITC significantly reduces CRC SP cells in vitro and impairs tumors growth in vivo. The combination of 5FU and PEITC led to synergistic cytotoxic effects against CRC cells in vitro and in vivo. Taken together, our data suggest that a GSH-mediated reduction in cellular ROS levels is an essential regulator of CRC SP cells mediated by the CD44v-xCT axis, and disrupting the redox status may eliminate the chemotherapy-resistant CRC SP cells with potentially significant benefits for cancer treatment.

Background and Aim: Lipopolysaccharide(LPS) could promote the progression of colorectal cancer, but the specific regulatory mechanisms are largely unknown. So, this study aim to clarify the mechanisms that LPS upregulated VEGFR-3, which promotes colorectal cancer cells migration and invasion with a mechanism of increased NF-#x03BA;B bind to the promoter of VEGFR-3. Methods: The present study examined the VEGFR-3 expression in colorectal cancer tissues and analyzed the relationship between the VEGFR-3 expression with clinical parameters. PCR, Western blot, CCK-8, colone formation assay, and Transwell assay detected that LPS promoted the migration and invasion and the role of VEGFR-3 in the process of colorectal carcinoma in vitro. Used the methods of promoter analysis, EMSA assay and ChIP assay to explore the mechanisms LPS increased the expression of VEGFR-3. Results: VEGFR-3 was significantly high expression in the colorectal cancer tissues. And the high expression was associated with the TNM stage and lymph node metastasis of colorectal cancer. LPS could promote the migration and invasion, which could be blocked by the neutralizing antibody IgG of VEGFR-3. And found that -159 nt to +65 nt was the crucial region of VEGFR-3 promoter. And detected that the NF-#x03BA;B was important transcription factor for the VEGFR-3 promoter. And LPS could increase NF-#x03BA;B binding to VEGFR-3 promoter and upregulated the expression of VEGFR-3 to exert biological functions. Conclusion: We have elucidated the relationship between LPS and the VEGFR-3 expression and revealed that VEGFR-3 play very important role in the process of LPS promoting the migration and invasion of colorectal cancer cells. Further illuminated the mechanism that LPS upregulated VEGFR-3 expression via increased NF-#x03BA;B bind to the promoter of VEGFR-3.
Cell Physiol Biochem 2016;39:1665-1678
Thursday, September 15, 2016 11:42 PM|Jin Luo, Yan-Ni Li, Fei Wang, Wei-Ming Zhang, Xin Geng|International Journal of Biological Sciences|Labels: Myc, RAS, CRC, gastric

A global DNA hypomethylation might activate oncogene transcription, thus promoting carcinogenesis and tumor development. S-Adenosylmethionine (SAM) serves as a major methyl donor in biological transmethylation events. The object of this study is to explore the influence of SAM on the status of methylation at the promoter of the oncogenes c-myc, H-ras and tumor-suppressor gene p16 (INK4a), as well as its inhibitory effect on cancer cells. The results indicated that SAM treatment inhibited cell growth in gastric cancer cells and colon cancer cells, and the inhibition efficiency was significantly higher than that in the normal cells. Under standard growth conditions, C-myc and H-ras promoters were hypomethylated in gastric cancer cells and colon cancer cells. SAM treatment resulted in a heavy methylation of these promoters, which consequently downregulated mRNA and protein levels. In contrast, there was no significant difference in mRNA and protein levels of p16 (INK4a) with and without SAM treatment. SAM can effectively inhibit the tumor cells growth by reversing the DNA hypomethylation on promoters of oncogenes, thus down-regulating their expression. With no influence on the expression of the tumor suppressor genes, such as P16, SAM could be used as a potential drug for cancer therapy.

Thursday, September 15, 2016 7:51 PM|Xue-hui Zhang, Hong-liang Yu, Fu-jing Wang, Yong-long Han, Wei-liang Yang|International Journal of Medical Sciences|Labels: CRC

Tumor cells have higher rates of glucose uptake and aerobic glycolysis to meet energy demands for proliferation and metastasis. The characteristics of increased glucose uptake, accompanied with aerobic glycolysis, has been exploited for the diagnosis of cancers. Although much progress has been made, the mechanisms regulating tumor aerobic glycolysis and energy production are still not fully understood. Here, we demonstrate that Pim-2 is required for glycolysis and energy production in colorectal tumor cells. Our results show that Pim-2 is highly expressed in colorectal tumor cells, and may be induced by nutrient stimulation. Activation of Pim-2 in colorectal cells led to increase glucose utilization and aerobic glycolysis, as well as energy production. While knockdown of Pim-2 decreased energy production in colorectal tumor cells and increased their susceptibility to apoptosis. Moreover, the effects of Pim-2 kinase on aerobic glycolysis seem to be partly dependent on mTORC1 signaling, because inhibition of mTORC1 activity reversed the aerobic glycolysis mediated by Pim-2. Our findings suggest that Pim-2-mediated aerobic glycolysis is critical for monitoring Warburg effect in colorectal tumor cells, highlighting Pim-2 as a potential metabolic target for colorectal tumor therapy.

Thursday, September 15, 2016 7:42 AM|Ehrensberger, S. H, Imaizumi, N, Monnier-;Benoit, S, Nichita, C, Myung, S.-;J, Kim, J. S, Song, S. Y, Kim, T. I, van der Weg, B, Meier, R, Borovicka, J, Beglinger, C, Vallet, C, Maerten, P, Rüegg, C, Dorta, G.|JournalTOCs API - Clinical Cancer Research (27 articles)|Labels: CEA, CRC

Blood Test for Colorectal Cancer Detection
Ciarloni, L Ehrensberger, S. H, Imaizumi, N, Monnier-Benoit, S, Nichita, C, Myung, S.-J, Kim, J. S, Song, S. Y, Kim, T. I, van der Weg, B, Meier, R, Borovicka, J, Beglinger, C, Vallet, C, Maerten, P, Rüegg, C, Dorta, G.
Clinical Cancer Research, Vol. 22, No. 18 (2016) pp. 4604 - 4611
Purpose: A blood test for early detection of colorectal cancer is a valuable tool for testing asymptomatic individuals and reducing colorectal cancer&ndash;related mortality. The objective of this study was to develop and validate a novel blood test able to differentiate patients with colorectal cancer and adenomatous polyps (AP) from individuals with a negative colonoscopy. Experimental Design: A case&ndash;control, multicenter clinical study was designed to collect blood samples from patients referred for colonoscopy or surgery. Predictive algorithms were developed on 75 controls, 61 large AP (LAP) &ge;1 cm, and 45 colorectal cancer cases and independently validated on 74 controls, 42 LAP, and 52 colorectal cancer cases (23 stages I&ndash;II) as well as on 245 cases including other colorectal findings and diseases other than colorectal cancer. The test is based on a 29-gene panel expressed in peripheral blood mononuclear cells alone or in combination with established plasma tumor markers. Results: The 29-gene algorithm detected colorectal cancer and LAP with a sensitivity of 79.5% and 55.4%, respectively, with 90.0% specificity. Combination with the protein tumor markers carcinoembryonic antigen (CEA) and CYFRA21-2 resulted in a specificity increase (92.2%) with a sensitivity for colorectal cancer and LAP detection of 78.1% and 52.3%, respectively. Conclusions: We report the validation of a novel blood test, Colox&reg;, for the detection of colorectal cancer and LAP based on a 29-gene panel and the CEA and CYFRA21-1 plasma biomarkers. The performance and convenience of this routine blood test provide physicians a useful tool to test average-risk individuals unwilling to undergo upfront colonoscopy. Clin Cancer Res; 22(18); 4604&ndash;11. &copy;2016 AACR.

Thursday, September 15, 2016 7:42 AM|W. M.|JournalTOCs API - Clinical Cancer Research (27 articles)|Labels: CRC, clinical trial

WRN Methylation Does Not Improve Irinotecan-Treated CRC Outcome
Bosch, L. J. W Luo, Y, Lao, V. V, Snaebjornsson, P, Trooskens, G, Vlassenbroeck, I, Mongera, S, Tang, W, Welcsh, P, Herman, J. G, Koopman, M, Nagtegaal, I. D, Punt, C. J. A, van Criekinge, W, Meijer, G. A, Monnat, R. J, Carvalho, B, Grady, W. M.
Clinical Cancer Research, Vol. 22, No. 18 (2016) pp. 4612 - 4622
Purpose: WRN promoter CpG island hypermethylation in colorectal cancer has been reported to increase sensitivity to irinotecan-based therapies. We aimed to characterize methylation of the WRN promoter, determine the effect of WRN promoter hypermethylation upon expression, and validate a previous report that WRN promoter hypermethylation predicts improved outcomes for patients with metastatic colorectal cancer (mCRC) treated with irinotecan-based therapy. Experimental Design: WRN methylation status was assessed using methylation-specific PCR and bisulfite sequencing assays. WRN expression was determined using qRT-PCR and Western blotting. WRN methylation status was correlated with overall survival (OS) and progression-free survival (PFS) in 183 patients with mCRC. Among these patients, 90 received capecitabine monotherapy as first-line therapy, and 93 received capecitabine plus irinotecan (CAPIRI) therapy as part of the CAIRO phase III clinical trial. Results: WRN mRNA and WRN protein expression levels were low in colorectal cancer cell lines and in primary colorectal cancer and were largely independent of WRN methylation status. Patients with methylated WRN colorectal cancer had a shorter OS compared with patients who had unmethylated WRN colorectal cancer (HR = 1.6; 95% confidence interval [CI], 1.2&ndash;2.2; P = 0.003). Patients with unmethylated WRN showed a significantly longer PFS when treated with CAPIRI compared with capecitabine alone (HR = 0.48; 95% CI, 0.32&ndash;0.70; P = 0.0001). In contrast, patients did not benefit from adding irinotecan to capecitabine when WRN was methylated (HR = 1.1; 95% CI, 0.69&ndash;1.77; P = 0.7). Conclusions: WRN expression is largely independent of WRN promoter hypermethylation in colorectal cancer. Moreover, we could not validate the previous finding that WRN promoter hypermethylation predicts improved clinical outcomes of mCRC treated with irinotecan-based therapy and found instead the opposite result. Clin Cancer Res; 22(18); 4612&ndash;22. &copy;2016 AACR.

Thursday, September 15, 2016 6:45 AM|Limagne, E., Euvrard, R., Thibaudin, M., Rebe, C., Derangere, V., Chevriaux, A., Boidot, R., Vegran, F., Bonnefoy, N., Vincent, J., Bengrine–Lefevre, L., Ladoire, S., Delmas, D., Apetoh, L., Ghiringhelli, F.|Cancer Research current issue|Labels: PD-1/PD-L1, CRC
Host immunity controls the development of colorectal cancer, and chemotherapy used to treat colorectal cancer is likely to recruit the host immune system at some level. Athough preclinical studies have argued that colorectal cancer drugs, such as 5-fluorouracil (5-FU) and oxaliplatin, exert such effects, their combination as employed in the oncology clinic has not been evaluated. Here, we report the results of prospective immunomonitoring of 25 metastatic colorectal cancer (mCRC) patients treated with a first-line combination regimen of 5-FU, oxaliplatin, and bevacizumab (FOLFOX–bevacizumab), as compared with 20 healthy volunteers. Before this therapy was initiated, T regulatory cells (Treg), Th17, and granulocytic myeloid-derived suppressor cells (gMDSC) were increased significantly in mCRC, but only a high level of gMDSC was associated with a poor prognosis. Chemotherapy modulated the Treg/Th17 balance by decreasing Treg and increasing Th17 cell frequency by 15 days after the start of treatment. Increased Th17 frequency was associated with a poor prognosis. FOLFOX–bevacizumab treatment elicited a decrease in gMDSC in 15 of 25 patients and was associated with a better survival outcome. Notably, the gMDSCs that expressed high levels of PD-L1, CD39, and CD73 exerted a robust immunosuppressive activity, relative to other myeloid cells present in blood, which could be reversed by blocking the CD39/CD73 and PD-1/PD-L1 axes. Our work underscores the critical prognostic impact of early modifications in Th17 and gMDSC frequency in mCRC. Furthermore, it provides a clinical rationale to combine FOLFOX–bevacizumab chemotherapy with inhibitors of ATP ectonucleotidases and/or anti-PD-1/PD-L1 antibodies to more effectively treat this disease. Cancer Res; 76(18); 5241–52. ©2016 AACR.
Wednesday, September 14, 2016 10:11 PM|Christoph Wilmanns, Sandra Steinhauer, Joachim Grossmann, Günther Ruf|International Journal of Biological Sciences|Labels: CRC

The purpose was to develop a metastatic score specific to the hepatic and peritoneal site in colorectal cancer patients from clinical, pathohistological and molecular markers potentially reflecting oncogenic activation (OA) or epithelial-mesenchymal transition (EMT), where OA may reflect an activation and EMT the functional loss of certain genes. The primary tumour stage (OA, EMT), lymphonodal stage (OA), the presence of a lymphangiosis carcinomatosa (OA), histological grade (OA, EMT), and immunoblot extraction of E-cadherin (OA, EMT) were differentially rated with zero to one or two points due to their potential contribution to each process and the resulting scores were validated in 27 colorectal cancer patients (three patients with pre-malignant adenomas, 16 with primaries and two with local recurrencies, three of which were metastatic to the peritoneum, six metastatic to the liver and two metastatic to both, the liver and the peritoneum, and five with hepatic secondaries, one of which at histology was metastatic to the peritoneum too). As a single parameter only the N-stage significantly contributed to OA (p<0.05). Median OA and EMT scores, however, were 3.5 and 2 in the case of primaries without further spread, 5 and 4 in those nodal positive, 5 and 4 in the case of peritoneal implants, 6 and 2 in the case of liver metastases, and 6.5 and 3 in the case of a simultaneous hepatic and peritoneal spread, respectively. These differences were significant when scores from patients with and without liver metastases (OA, p<0.002) or with peritoneal implants and isolated hepatic spread (EMT, p<0.01) were compared. The results suggest a site-specific contribution of OA and EMT to tumour progression in human colon cancer.

Contributors : Ran Afik ; Ehud Zigmond ; Milena Vugman ; Metsada Pasmanik-Chor ; Anjana Shenoy ; Elad Bassat ; Zamir Halpern ; Tamar Geiger ; Irit Sagi ; Chen Varol
Series Type : Expression profiling by array
Organism : Mus musculus

Ly6Chi monocytes massively infiltrate the CRC-tumors by virtue of their CCR2 expression and further mature into Ly6CloF4/80hi CD64hiMHCII+ TAM upon tumor progression. We demonstrated that TAM-deficient tumors display impaired tumor-growth via alternation of the ECM morphology, structure and composition. Using advanced high-resolution optical imaging to visualize the tumoral-ECM macromolecule network together with transcriptomic and proteomic approaches we unraveled that TAM play critical role in the deposition, linearization and cross-linking of collagenous ECM. Remarkably, we show that cues embedded in ECM by TAM-mediated remodeling activity promote tumor cell proliferation in vitro and orthotopic tumor development in vivo.
Abnormal remodeling of the extracellular matrix (ECM) structural and compositional is a hallmark of many cancers. While tumor associated macrophages (TAM) are considered pivotal players in mounting pro-tumoral functions, their role in tumor-ECM remodeling remains largely ambiguous. We found that TAM-deficient CRC tumors established in Ccr2-/- mice exhibited attenuated growth. Advanced molecular imaging revealed that the enhanced deposition, linearization and cross-linking of collagen fibers typical to tumor growth were absent in the Ccr2-/- tumors. Integrating transcriptomic and proteomic approaches we defined a TAM signature of ECM remodeling enzymes, structural and affiliated proteins involved with the synthesis and assembly of collagen. The Ccr2-/- tumors displayed altered ECM composition with 348 proteins that were differentially expressed in comparison with WT tumors, among them 46 were ECM related. Decellularized 3D ECM fragments extracted from WT tumors, but not from Ccr2-/- tumors or upstream healthy colon, enhanced tumor cell proliferation in vitro and tumor development in the native colonic environment indicating that TAM have a critical role in priming pathological ECM cues.

Wednesday, September 14, 2016 5:53 PM|Maximilian J. Waldner, Sebastian Foersch, Markus F. Neurath|International Journal of Biological Sciences|Labels: CRC, IL

Growing evidence proposes an important role for pro-inflammatory cytokines during tumor development. Several experimental and clinical studies have linked the pleiotropic cytokine interleukin-6 (IL-6) to the pathogenesis of sporadic and inflammation-associated colorectal cancer (CRC). Increased IL-6 expression has been related to advanced stage of disease and decreased survival in CRC patients. According to experimental studies, these effects are mediated through IL-6 trans-signaling promoting tumor cell proliferation and inhibiting apoptosis through gp130 activation on tumor cells with subsequent signaling through Janus kinases (JAKs) and signal transducer and activator of transcription 3 (STAT3).

During recent years, several therapeutics targeting the IL-6/STAT3 pathway have been developed and pose a promising strategy for the treatment of CRC. This review discusses the molecular mechanisms and possible therapeutic targets involved in IL-6 signaling in CRC.

Wednesday, September 14, 2016 5:53 PM|Daijun Zhou, Qiang Mei, Han Luo, Bo Tang, Peiwu Yu|International Journal of Biological Sciences|Labels: DHFR, CRC

Polymorphisms in genes involved in folate metabolism may modulate the risk of colorectal cancer (CRC), but data from published studies are conflicting. The current meta-analysis was performed to address a more accurate estimation. A total of 41 (17,552 cases and 26,238 controls), 24(8,263 cases and 12,033 controls), 12(3,758 cases and 5,646 controls), and 13 (5,511 cases and 7,265 controls) studies were finally included for the association between methylenetetrahydrofolate reductase (MTHFR) C677T and A1289C, methione synthase reductase (MTRR) A66G, methionine synthase (MTR) A2756G polymorphisms and the risk of CRC, respectively. The data showed that the MTHFR 677T allele was significantly associated with reduced risk of CRC (OR = 0.93, 95%CI 0.90-0.96), while the MTRR 66G allele was significantly associated with increased risk of CRC (OR = 1.11, 95%CI 1.01-1.18). Sub-group analysis by ethnicity revealed that MTHFR C677T polymorphism was significantly associated with reduced risk of CRC in Asians (OR = 0.80, 95%CI 0.72-0.89) and Caucasians (OR = 0.84, 95%CI 0.76-0.93) in recessive genetic model, while the MTRR 66GG genotype was found to significantly increase the risk of CRC in Caucasians (GG vs. AA: OR = 1.18, 95%CI 1.03-1.36). No significant association was found between MTHFR A1298C and MTR A2756G polymorphisms and the risk of CRC. Cumulative meta-analysis showed no particular time trend existed in the summary estimate. Probability of publication bias was low across all comparisons illustrated by the funnel plots and Egger's test. Collectively, this meta-analysis suggested that MTHFR 677T allele might provide protection against CRC in worldwide populations, while MTRR 66G allele might increase the risk of CRC in Caucasians. Since potential confounders could not be ruled out completely, further studies were needed to confirm these results.

Wednesday, September 14, 2016 4:21 PM|Fred Hutchinson Cancer Research Center/UW Medicine Cancer Consortium - Clinical Trials|Labels: CRC, clinical trial
The purpose of this study is to determine if the True Human Monoclonal antibody Xilonix (MABp1) can prolong the life of colorectal carcinoma patients that are refractory to standard therapy.
Wednesday, September 14, 2016 11:10 AM|Russell C. Langan, John E. Mullinax, Satyajit Ray, Manish T. Raiji, Nicholas Schaub, Hong-Wu Xin, Tomotake Koizumi, Seth M. Steinberg, Andrew Anderson, Gordon Wiegand, Donna Butcher, Miriam Anver, Anton J. Bilchik, Alexander Stojadinovic, Udo Rudloff, Itzhak Avital|Journal of Cancer|Labels: CRC, biomarker diagnostic

Introduction: Over 50% of patients with colorectal cancer (CRC) will progress and/or develop metastases. Biomarkers capable of predicting progression, risk stratification and therapeutic benefit are needed. Cancer stem cells are thought to be responsible for tumor initiation, dissemination and treatment failure. Therefore, we hypothesized that CRC cancer stem cell markers (CRCSC) will identify a group of patients at high risk for progression.

Methods: Paraffin-embedded tissue cores of normal (n=8), and histopathologically well-defined primary (n= 30) and metastatic (n=10) CRC were arrayed in duplicate on tissue microarrays (TMAs). Expression profiles of non-CD133 CRCSC (CD29, CD44, ALDH1A1, ALDH1B1, EpCam, and CD166) were detected by immunohistochemistry and the association with clinicopathological data and patient outcomes was determined using standard statistical methodology. An independent pathologist, blinded to the clinical data scored the samples. Scoring included percent positive cells (0 to 4, 0 = <10%, 1 = 10 - 24%, 2 = 25 - 49%, 3 = 50 - 74%, 4 = 75 - 100%), and the intensity of positively stained cells (0 to 4; 0 = no staining, 1 = diminutive intensity, 2 = low intensity, 3 = intermediate intensity, 4 = high intensity). The pathologic score represents the sum of these two values, reported in this paper as a combined IHC staining score (CSS).

Results: Of 30 patients 7 were AJCC stage IIA, 10 stage IIIB, 7 stage IIIC and 6 stage IV. Median follow-up was 113 months. DFI was 17 months. Median overall survival (OS) was not reached. Stage-specific OS was: II - not reached; III - not reached; IV - 11 months. In a univariate analysis, poor OS was associated with loss of CD29 expression; median OS, 32 months vs. not reached for CSS 3-7 vs. >7.5, respectively; p=0.052 comparing entire curves, after adjustment. In a Cox model analysis, loss of CD29 exhibited a trend toward association with survival (p=0.098) after adjusting for the effect of stage (p=0.0076). Greater expression of ALDH1A1 was associated with increasing stage (p=0.042 over stages 2, 3b, 3c, and 4) while loss of CD29 expression exhibited a trend toward being associated with stages 3 and 4 (p=0.08). Compared to normal colon tissue, primary tumors were associated with increased expression of ALDH1B1 (p=0.008). ALD1H1B1 expression level differed according to whether the tumor was moderately or poorly differentiated, well differentiated, or mucinous; the highest expression levels were associated with moderately or poorly differentiated tumors (p=0.011). Lymph node metastases were associated with a trend toward decreased expression of EpCAM (p = 0.06) when comparing 0 vs. 1 vs. 2+ positive lymph nodes, as was CD29 (p = 0.08) when comparing 0 vs. any positive lymph nodes. Compared to normal colon tissue metastatic colon cancers from different patients were associated with increased ALDH1B1 expression (p=0.001) whereas CD29 expression was higher in normal colonic tissue (p=0.014).

Conclusion: CD29 may be associated with survival as well as clinical stage and number of lymph nodes. ALDH1B1 expression was associated with differentiation as well as type of tissue evaluated. ALDH1A1 was associated with clinical stage, and decreased EpCAM expression was found in patients with advanced lymph node stage. CRCSCs may be useful biomarkers to risk stratify, and estimate outcomes in CRC. Larger prospective studies are required to validate the current findings.

Wednesday, September 14, 2016 7:55 AM|KISS, I., MLCOCHOVA, J., BORTLICEK, Z., POPRACH, A., DRABEK, J., VYCHYTILOVA-FALTEJSKOVA, P., SVOBODA, M., BUCHLER, T., BATKO, S., RYSKA, A., HAJDUCH, M., SLABY, O.|Anticancer Research current issue|Labels: CRC

Background: In metastatic colorectal cancer (mCRC), panitumumab is generally considered to be ineffective after the progression on cetuximab therapy. However, few studies have demonstrated that a small subset of mCRC patients may benefit from panitumumab in this setting. Patients and Methods: In our study, wild-type KRAS mCRC patients, enrolled into the nationwide Czech registry CORECT between January 2007 and December 2012, were screened for panitumumab therapy after progression on cetuximab. Results: We identified 26 mCRC in the registry with well documented progression on cetuximab in combination with irinotecan-based chemotherapy (FOLFIRI or irinotecan alone) who received panitumumab monotherapy. Partial response (PR) was achieved in 3 (11.5%) patients and stable disease (SD) in 7 (26.9%) patients after 8 weeks of therapy. Thirteen (50.0%) patients had evidence of progressive disease (PD) and in 3 (11.5%) cases response was not available. Furthermore, we confirmed that higher expression levels of newly described biomarker, miR-31-5p, in tumor are significantly associated with shorter progression-free survival (PFS) in patients treated with cetuximab (p=0.038); however, we did not observe association between miR-31-5p and response to panitumumab in mCRC patients after progression on cetuximab. Conclusion: It remains possible that a subset of mCRC patients may benefit from panitumumab after progression on cetuximab.

Wednesday, September 14, 2016 7:55 AM|RACHAR, V., CZEJKA, M., KITZMUELLER, M., BUCHNER, P., LICHTNECKERT, M., GREIL, R., GEILER, H., DITTRICH, C.|Anticancer Research current issue|Labels: CRC, clinical trial

Aim: This study focuses on the plasma disposition and metabolic activation of capecitabine (CCB) when administered alone or when combined with cetuximab (CTX). Patients and Methods: Twenty-four chemo-naïve patients with KRAS wild-type colorectal cancer were randomized into two arms and received either CCB alone (1,000 mg/m2 bid p.o.), followed by CCB plus CTX (loading dose (LD)=400 mg/m2 followed by 250 mg/m2 weekly i.v. maintenance dose) (Arm A; n=12 patients (patients)) or CCB plus CTX followed by CCB alone (Arm B; n=12 patients). Plasma samples were collected from the cubital vein and CCB, 5’-desoxy-5-fluorocytidine (5’-DFCR) and 5’-desoxy-5 fluorouridine (5’-DFUR) were quantified by a sensitive, selective reversed phase high-performance liquid chromatography (HPLC) assay. Non-compartment pharmacokinetic parameters have been calculated by Phoenix WinNonlin. Results: No clinically relevant impact of CTX on CCB pharmacokinetic parameters and metabolic conversion could be detected in both arms after statistical evaluation (ANOVA). Conclusion: From the pharmacokinetic point of view, co-administration of CTX to CCB seems to be safe.

Wednesday, September 14, 2016 7:55 AM|LING, E., RINGEL, A., SIGAL-BATIKOFF, I., ABU-FREHA, N., VAKNINE, H., FRIAH, W., RESHEF, A., PINSK, I., FICH, A., LAMPRECHT, S.|Anticancer Research current issue|Labels: CRC

Background/Aim: Cancer-associated fibroblasts (CAFs) play an important role in tumor development and progression. The prevailing consensus favors the view that a specific epigenetic signature underpins the stable CAF phenotype. The aim of the present study was to assess global DNA methylation in CAFs during the adenoma–carcinoma sequence in non-familial sporadic human colorectal cancer (CRC). Patients and Methods: Immunohistochemical staining of nuclear 5-methylcytosine (5’-meCyt) was performed in matched samples of colonic tumor tissue and normal colonic mucosa excised from six patients with adenomas and four with adenocarcinomas. The staining intensity was expressed semi-quantitatively as the immunohistochemical staining score (ISS). Results: ISS values of human colonic CAFs and adenomatous samples were 14.00±2.2 and 14.08±1.8, respectively, showing no statistically significant difference. In contrast, a marked trend was found towards global DNA hypomethylation in CAFs from adenocarcinomatous specimens compared to matched normal mucosa: ISS: 9.25±2.44 (range=6-11) vs. 16.17±0.75, respectively, p<0.03. Conclusion: Final stages of cancer development in CRC are associated with global DNA hypomethylation in stromal CAFs.

Wednesday, September 14, 2016 7:55 AM|DA SILVA, F. C., WERNHOFF, P., DOMINGUEZ-BARRERA, C., DOMINGUEZ-VALENTIN, M.|Anticancer Research recent issues|Labels: CRC

In the past two decades, significant advances have been made in our understanding of colorectal (CRC) tumors with DNA mismatch (MMR) repair deficiency. The knowledge from molecular and genetic alterations in a variety of clinical conditions has refined the disease terminology and classification. Hereditary non-polyposis colorectal cancer (HNPCC) encompasses a spectrum of conditions that have significant phenotypic overlapping that makes clinical diagnosis a challenging task. Distinguishing among the HNPCC disorders is clinically important, as the approach to surveillance for patients and their at-risk family members differs according to risks for colonic and extracolonic cancer associated with each syndrome. Prospective and next-generation studies will provide valuable clinical information regarding the natural history of disease that will help differentiate the Lynch syndrome mimics and guide diagnosis and management for heterogeneous conditions currently grouped under the category of familial CRC. The review is intended to present and discuss the molecular nature of various conditions related to MMR deficiency and discusses the tools and strategies that have been used in detecting these conditions.

Wednesday, September 14, 2016 7:55 AM|STANKEVICIUS, V., VASAUSKAS, G., NOREIKIENE, R., KUODYTE, K., VALIUS, M., SUZIEDELIS, K.|Anticancer Research current issue|Labels: CRC

Background: Cancer cells grown in a 3D culture are more resistant to anticancer therapy treatment compared to those in a monolayer 2D culture. Emerging evidence has suggested that the key reasons for increased cell survival could be gene expression changes in cell–extracellular matrix (ECM) interaction-dependent manner. Materials and Methods: Global gene-expression changes were obtained in human colorectal carcinoma HT29 and DLD1 cell lines between 2D and laminin-rich (lr) ECM 3D growth conditions by gene-expression microarray analysis. The most significantly altered functional categories were revealed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Results: The microarray data revealed that 841 and 1190 genes were differentially expressed in colorectal carcinoma DLD1 and HT29 cells. KEGG analysis indicated that the most significantly altered categories were cell adhesion, mitogen-activated protein kinase and immune response. Conclusion: Our results indicate altered pathways related to cancer development and progression and suggest potential ECM-regulated targets for the development of anticancer therapies.

Wednesday, September 14, 2016 7:55 AM|HOLUBEC, L., POLIVKA, J., SAFANDA, M., KARAS, M., LISKA, V.|Anticancer Research current issue|Labels: EGFR, CRC

Monoclonal antibodies binding the epidermal growth factor receptor (EGFR), such as cetuximab or panitumumab, are widely used targeted therapeutics for the treatment of patients with colorectal cancer. The clinical significance of these drugs has so far been associated with combined chemotherapy or radiation. It has been shown that these treatment strategies have their clinical limitations and do not fully exploit the immunomodulatory effect of these drugs. In this review, we discuss the mechanisms of immunomodulation together with the anticancer immune response to the monoclonal antibodies targeted to the EGFR. The combination of anti-EGFR monoclonal antibodies with other immunotherapeutic treatment modalities certainly brings new opportunities for targeted therapy in patients with colorectal cancer.

Tuesday, September 13, 2016 11:38 PM|Shiva Basnet, Zhen-yu Zhang, Wen-qiang Liao, Shu-heng Li, Ping-shu Li, Hai-yan Ge|Journal of Cancer|Labels: CRC

Background: Circulating cell-free DNA (cfDNA) is a promising candidate biomarker for detection, monitoring and survival prediction of colorectal cancer (CRC). However, its prognostic significance for patients with CRC remains controversial. To derive a precise estimation of the prognostic significance of cfDNA, a meta-analysis was performed.

Methods: We made a systematic search in data base of the Science Citation Index Embase and Pubmed for studies reporting prognostic data of cfDNA in CRC patients. The data of cfDNA on recurrences-free survival (RFS) and overall survival (OS) were extracted and measured in hazard rates (HRs) and 95% confident intervals (CIs). Subgroup analyses were carried out as well. Finally, the meta-analysis is accompanied with nine studies including 19 subunits.

Results: The pooled HRs with 95% CIs revealed strong associations between cfDNA and RFS (HR [95%CI]=2.78[2.08-3.72], I2=32.23%, n=7) along with OS (HR [95%CI]=3.03[2.51-3.66], I2=29.24%, n=12) in patients with CRC. Entire subgroup analyses indicated strong prognostic value of cfDNA irrespective tumor stage, study size, tumor markers, detection methods and marker origin.

Conclusions: All the results exhibits that appearance of cfDNA in blood is an indicator for adverse RFS and OS in CRC patients.

Tuesday, September 13, 2016 11:38 PM|Yumin Wang, Dan Xue, Yuwei Li, Xuya Pan, Xueying Zhang, Biao Kuang, Ming Zhou, Xiaoling Li, Wei Xiong, Guiyuan Li, Zhaoyang Zeng, Tubao Yang|Journal of Cancer|Labels: CRC, biomarker diagnostic

Background: MALAT-1 is significantly overexpressed in various cancers, suggesting that it might be a potential biomarker of cancer.

Methods: A meta-analysis was performed using microarray data obtained via the Affymetrix Human Genome U133 Plus 2.0 platform found in the Gene Expression Omnibus (GEO) database and data obtained through a systematic search of PubMed and Web of Science. The pooled odds ratio (OR) and hazard ratio (HR) with 95% CI (Confidence interval) were used to judge the value of biomarkers.

Results: A total of 28 studies were included in this meta-analysis, comprising a total of 3573 patients. MALAT-1 was significantly linked with over survival (OS) (HR=1.58, 95%CI: 1.12-2.23), recurrence-free survival (RFS) (HR=2.32, 95% CI: 1.68-3.19) and death-free survival (DFS) (HR=3.28, 95% CI: 1.52-7.09). We found that MALAT-1 was a risk factor in the prognoses of lung cancer (HR=1.54, 95%CI: 1.01-2.34), digestive system cancer (HR=2.16, 95% CI: 1.34-3.48) and ovarian cancer (HR=3.98, 95% CI: 1.54-10.25). In contrast, MALAT-1 was a safe factor in the prognosis of B cell lineage cancer (HR=0.45, 95% CI: 0.33-0.61). MALAT-1 was also a risk factor of RFS in breast cancer (HR=1.97, 95% CI: 1.25-3.09) and the TNM stage in pancreatic cancer (OR=3.65, 95% CI: 1.86-7.18) and glioma (OR=4.30, 95% CI: 1.90-9.73) and was a safe factor in colorectal cancer (OR=0.17, 95% CI: 0.08-0.35). MALAT-1 was significantly associated with lymph node metastasis in clear cell carcinoma (OR=5.04, 95% CI: 2.36-10.78) and distant metastasis in pancreatic cancer (OR=11.64, 95% CI: 2.13-63.78).

Conclusions: MALAT-1 can serve as a molecular marker in different types of cancers.

Tuesday, September 13, 2016 11:38 PM|Jianming He, Xi Liang, Fen Luo, Xuedan Chen, Xueqing Xu, Fengchao Wang, Zhenping Zhang|Journal of Cancer|Labels: caspase, P53, CRC

Three-dimensional (3D) culture models represent a better approximation of solid tumor tissue architecture, especially cell adhesion, in vivo than two-dimensional (2D) cultures do. Here, we explored the role of architecture in chemosensitivity to platinum in colon cancer. Under the 3D culture condition, colon cancer cells formed multicellular spheroids, consisting of layers of cells. 3D cultures displayed significantly decreased sensitivity to platinum compared with 2D cultures. Platinum increased p53 in a dose-dependent and time-dependent manner. There was no detectable difference in basal p53 levels between 3D cultures and 2D cultures but cisplatin induced less p53 in both HCT116 3D cultures and LoVo 3D cultures. It was not due to cisplatin concentration because cisplatin induced similar γ-H2AX in 3D vs 2D. Knockdown of p53 significantly decreased sensitivity to platinum in 3D cultures. Knockdown of p53 decreased cleaved caspase 3 and apoptosis induced by cisplatin. These findings indicate that 3D architecture confers decreased chemosensitivity to platinum and p53 is involved in the mechanism. Knockdown of p53 decreased cisplatin's induction of c-Jun N-terminal kinase 1/2 (JNK1/2) activation, whereas inhibition of JNK1/2 activation increased chemosensitivity. Inhibition of p38 activation decreased cisplatin's induction of p53, but no difference in p38 activation by cisplatin was observed between 2D cultures and 3D cultures. Taken together, our results suggest that p53 is involved in a 3D architecture-mediated decrease in chemosensitivity to platinum in colon cancer. Mitogen-activated protein kinases (JNK1/2 and p38) do not play a dominant role in the mechanism.

Tuesday, September 13, 2016 11:38 PM|Yaping Zhang, Chengyan He, Ling Qiu, Yanmin Wang, Xuzhen Qin, Yujie Liu, Zhili Li|Journal of Cancer|Labels: CRC, biomarker diagnostic

Background: To screen biomarkers to differentiate early-stage colorectal cancer (CRC) from benign colorectal disease (BCD) and healthy controls.

Materials & Methods: Quantitative and qualitative analysis of C16:1, C18:3, C18:2, C18:1, C20:4, and C22:6 in 185 healthy controls, 55 patients with BCD, and 139 patients with CRC was performed. Comparisons of their levels in between CRC patients, BCD patients, and healthy controls were performed using Mann-Whitney U test.

Results: Serum levels of C16:1, C18:3, C18:2, C18:1, C20:4, and C22:6 in CRC patients were significantly decreased compared with healthy controls and BCD patients. A combination of C16:1, C18:2, C20:4, and C22:6 has excellent diagnostic performance to differentiate early-stage CRC patients from healthy controls plus BCD patients, with an AUC of 0.926, a sensitivity of 84.6%, and a specificity of 89.8%.

Conclusions: Serum levels of C16:1, C18:2, C20:4, and C22:6 could be diagnostic indicators of early-stage CRC patients.

Tuesday, September 13, 2016 11:38 PM|Kenichi Maeda, Chiemi Saigo, Yusuke Kito, Takuji Sakuratani, Kazuhiro Yoshida, Tamotsu Takeuchi|Journal of Cancer|Labels: CRC

Recent research advances highlighted an intestinal goblet cell-produced lectin, intelectin-1 (also known as omentin-1), as a tumor suppressor. One study indicated that downregulation of intelectin-1 may be related to the unfavorable prognosis among patients with colorectal carcinoma at an advanced stage. The present study was aimed at analyzing the expression of a hitherto uncharacterized transmembrane protein TMEM207 in colorectal carcinoma, and we found that the TMEM207 function is linked to intelectin-1 processing. With specific antibodies, TMEM207 immunoreactivity was detected in 38 of 216 colorectal cancer tissue samples. TMEM207 immunoreactivity correlated inversely with lymph node metastatic status (p < 0.01). TMEM207 expression significantly correlated with the mucinous phenotype of colorectal carcinoma. A coimmunoprecipitation assay revealed an interaction between intelectin-1 and TMEM207 in colorectal cancer cells. A proximal ligation assay indicated that intelectin-1 and TMEM207 were colocalized to the cytoplasm of the colorectal cancer cells. A small-interfering-RNA-mediated knockdown of TMEM207 increased polyubiquitination and proteasome degradation of intelectin-1 in cultured colorectal cancer cells and decreased intelectin-1 secretion. These findings indicate that a loss of TMEM207 expression leads to insufficient intelectin-1 production thus promoting colorectal carcinogenesis.

Tuesday, September 13, 2016 11:38 PM|Kamal Datta, Shubhankar Suman, Santosh Kumar, Albert J Fornace|Journal of Cancer|Labels: proteasome, WNT, CRC

Adult colorectal epithelium undergoes continuous renewal and maintains homeostatic balance through regulated cellular proliferation, differentiation, and migration. The canonical Wnt signaling pathway involving the transcriptional co-activator β-catenin is important for colorectal development and normal epithelial maintenance, and deregulated Wnt/β-catenin signaling has been implicated in colorectal carcinogenesis. Colorectal carcinogenesis has been linked to radiation exposure, and radiation has been demonstrated to alter Wnt/β-catenin signaling, as well as the proteasomal pathway involved in the degradation of the signaling components and thus regulation of β-catenin. The current review discusses recent progresses in our understanding of colorectal carcinogenesis in relation to different types of radiation and roles that radiation quality plays in deregulating β-catenin and ubiquitin-proteasome pathway (UPP) for colorectal cancer initiation and progression.

Tuesday, September 13, 2016 10:05 PM|Mo, A., Jackson, S., Varma, K., Carpino, A., Giardina, C., Devers, T. J., Rosenberg, D. W.|Molecular Cancer Research recent issues|Labels: BRAF, CRC

Although the progression of mutated colonic cells is dependent upon interactions between the initiated epithelium and surrounding stroma, the nature of these interactions is poorly understood. Here, the development of an ultrasensitive laser capture microdissection (LCM)/RNA-seq approach for studying the epithelial and stromal compartments of aberrant crypt foci (ACF) is described. ACF are the earliest identifiable preneoplastic lesion found within the human colon and are detected using high-definition endoscopy with contrast dye spray. The current analysis focused on the epithelium of ACF with somatic mutations to either KRAS, BRAF, or APC, and expression patterns compared with normal mucosa from each patient. By comparing gene expression patterns among groups, an increase in a number of proinflammatory NF-B target genes was identified that was specific to ACF epithelium, including TIMP1, RELA, and RELB. Distinct transcriptional changes associated with each somatic mutation were observed and a subset of ACF display BRAFV600E-mediated senescence-associated transcriptome characterized by increased expression of CDKN2A. Finally, LCM-captured ACF-associated stroma was found to be transcriptionally distinct from normal-appearing stroma, with an upregulation of genes related to immune cell infiltration and fibroblast activation. Immunofluorescence confirmed increased CD3+ T cells within the stromal microenvironment of ACF and an abundance of activated fibroblasts. Collectively, these results provide new insight into the cellular interplay that occurs at the earliest stages of colonic neoplasia, highlighting the important role of NF-B, activated stromal fibroblasts, and lymphocyte infiltration.

Implications: Fibroblasts and immune cells in the stromal microenvironment play an important role during the earliest stages of colon carcinogenesis. Mol Cancer Res; 14(9); 795–804. ©2016 AACR.

Tuesday, September 13, 2016 10:05 PM|Li, Q., Qin, Y., Wei, P., Lian, P., Li, Y., Xu, Y., Li, X., Li, D., Cai, S.|Molecular Cancer Research recent issues|Labels: CRC

Growth arrest–specific 1 (Gas1) plays a critical role in growth suppression. Previous study indicated that Gas1 was closely associated with survival in patients with colorectal cancer; however, the underlying molecular mechanism remains unclear. In this study, we sought to determine the role of Gas1 in tumorigenesis and metastasis, and elucidate the possible mechanism. First, Gas1 was determined as a negative regulator of oncogenesis and metastasis in colorectal cancer. Mechanistically, Gas1 negatively regulated the aerobic glycolysis, a process that contributed to tumor progression and metastasis by providing energy source and building blocks for macromolecule synthesis. To further consolidate the role of Gas1 in glycolysis, the impact of Gas1 in the transcription of key glycolytic enzymes for glucose utilization was examined. As expected, GLUT4, HK2, and LDHB exhibited a decreased expression pattern. Consistent with this observation, an in vivo subcutaneous xenograft mouse model also confirmed the hypothesis that Gas1 is a negative regulator of glycolysis as reflected by the decreased 18FDG uptake in PET/CT system. Moreover, Gas1 negatively regulated the AMPK/mTOR/p70S6K signaling axis, a well-established cascade that regulates malignant cancer cell behaviors including proliferation, metastasis, and aberrant cancer metabolism. In the end, it was determined that Gas1 is a transcriptional target of FOXM1, whose role in colorectal cancer has been widely studied. Taken together, these studies establish Gas1 as a negative regulator in colorectal cancer.

Implications: Gas1 suppresses cell proliferation, invasion, and aerobic glycolysis of colorectal cancer both in vitro and in vivo. Mechanistically, Gas1 inhibited EMT and the Warburg effect via AMPK/mTOR/p70S6K signaling, and Gas1 itself was directly regulated by the transcription factor FOXM1. Mol Cancer Res; 14(9); 830–40. ©2016 AACR.

Tuesday, September 13, 2016 10:05 PM|Cantarino, N., Musulen, E., Valero, V., Peinado, M. A., Perucho, M., Moreno, V., Forcales, S.-V., Douet, J., Buschbeck, M.|Molecular Cancer Research recent issues|Labels: CRC

Peptidyl arginine deiminases (PADI) are a family of enzymes that catalyze the poorly understood posttranslational modification converting arginine residues into citrullines. In this study, the role of PADIs in the pathogenesis of colorectal cancer was investigated. Specifically, RNA expression was analyzed and its association with survival in a cohort of 98 colorectal cancer patient specimens with matched adjacent mucosa and 50 controls from donors without cancer. Key results were validated in an independent collection of tumors with matched adjacent mucosa and by mining of a publicly available expression data set. Protein expression was analyzed by immunoblotting for cell lines or IHC for patient specimens that further included 24 cases of adenocarcinoma with adjacent dysplasia and 11 cases of active ulcerative colitis. The data indicate that PADI2 is the dominantly expressed PADI enzyme in colon mucosa and is upregulated during differentiation. PADI2 expression is low or absent in colorectal cancer. Frequently, this occurs already at the stage of low-grade dysplasia. Mucosal PADI2 expression is also low in ulcerative colitis. The expression level of PADI2 in tumor and adjacent mucosa correlates with differential survival: low levels associate with poor prognosis.

Implications: Downregulation of PADI2 is an early event in the pathogenesis of colorectal cancer associated with poor prognosis and points toward a possible role of citrullination in modulating tumor cells and their microenvironment. Mol Cancer Res; 14(9); 841–8. ©2016 AACR.

Tuesday, September 13, 2016 8:19 PM|K. Trumpi, B.L. Emmink, A.M. Prins, M.G.H. van Oijen, P.J. van Diest, C.J.A. Punt, M. Koopman, O. Kranenburg, I.H.M. Borel Rinkes|Journal of Cancer (RSS 2.0)|Labels: P-gp, CRC

Background: Active efflux of irinotecan by ATP-binding cassette (ABC)-transporters, in particular ABCB1 and ABCG2, is a well-established drug resistance mechanism in vitro and in pre-clinical mouse models, but its relevance in colorectal cancer (CRC) patients is unknown. Therefore, we assessed the association between ABC-transporter expression and tumour response to irinotecan in patients with metastatic CRC.

Methods: Tissue microarrays of a large cohort of metastatic CRC patients treated with irinotecan in a prospective study (CAIRO study; n=566) were analysed for expression of ABCB1 and ABCG2 by immunohistochemistry. Kaplan-Meier and Cox proportional hazard regression analyses were performed to assess the association of ABC transporter expression with irinotecan response. Gene expression profiles of 17 paired tumours were used to assess the concordance of ABCB1/ABCG2 expression in primary CRC and corresponding metastases.

Results: The response to irinotecan was not significantly different between primary tumours with positive versus negative expression of ABCB1 (5.8 vs 5.7 months, p=0.696) or ABCG2 (5.7 vs 6.1 months, p=0.811). Multivariate analysis showed neither ABCB1 nor ABCG2 were independent predictors for progression free survival. There was a mediocre to poor concordance between ABC-transporter expression in paired tumours.

Conclusion: In metastatic CRC, ABC-transporter expression in the primary tumour does not predict irinotecan response.

Tuesday, September 13, 2016 8:19 PM|Ammara Abdullah, Sanam Sane, Jessica L Freeling, Hongmin Wang, Dong Zhang, Khosrow Rezvani|Journal of Cancer (RSS 2.0)|Labels: P53, CRC, IL

The subcellular localization, expression level, and activity of anti-cancer proteins alter in response to intrinsic and extrinsic cellular stresses to reverse tumor progression. The purpose of this study is to determine whether UBXN2A, an activator of the p53 tumor suppressor protein, has different subcellular compartmentalization in response to the stress of DNA damage. We measured trafficking of the UBXN2A protein in response to two different DNA damage stresses, UVB irradiation and the genotoxic agent Etoposide, in colon cancer cell lines. Using a cytosol-nuclear fractionation technique followed by western blot and immunofluorescence staining, we monitored and quantitated UBXN2A and p53 proteins as well as p53's downstream apoptotic pathway.

We showed that the anti-cancer protein UBXN2A acts in the early phase of cell response to two different DNA damage stresses, being induced to translocate into the cytoplasm in a dose- and time-dependent manner. UVB-induced cytoplasmic UBXN2A binds to mortalin-2 (mot-2), a known oncoprotein in colon tumors. UVB-dependent upregulation of UBXN2A in the cytoplasm decreases p53 binding to mot-2 and activates apoptotic events in colon cancer cells. In contrast, the shRNA-mediated depletion of UBXN2A leads to significant reduction in apoptosis in colon cancer cells exposed to UVB and Etoposide. Leptomycin B (LMB), which was able to block UBXN2A nuclear export following Etoposide treatment, sustained p53-mot-2 interaction and had partially antagonistic effects with Etoposide on cell apoptosis. The present study shows that nucleocytoplasmic translocation of UBXN2A in response to stresses is necessary for its anti-cancer function in the cytoplasm. In addition, LMB-dependent suppression of UBXN2A's translocation to the cytoplasm upon stress allows the presence of an active mot-2 oncoprotein in the cytoplasm, resulting in p53 sequestration as well as activation of other mot-2-dependent growth promoting pathways.

Tuesday, September 13, 2016 8:19 PM|Jung Han Kim, Dae Young Zang, Ik-Joo Chung, Sang-Hee Cho, Keon Uk Park, Ho-Suck Oh, Kyung Hee Lee, Bong Hwa Lee, Min-Jeong Kim, Choong Kee Park, Boram Han, Hyeong Su Kim, Dae Ro Choi, Hun Ho Song, Joo Young Jung|Journal of Cancer (RSS 2.0)|Labels: CRC, clinical trial

Background: Capecitabine plus oxaliplatin (XELOX) is considered one of the primary chemotherapy regimens for patients with metastatic colorectal cancer (CRC). Oxaliplatin plus S-1 (OS) has also demonstrated significant efficacy in CRC. We performed this randomized phase II study to evaluate the efficacy and toxicity of XELOX versus OS as first-line chemotherapy in patients with metastatic CRC.

Methods: Patients were assigned randomly to receive either OS or XELOX chemotherapy. Oxaliplatin was administered intravenously to all patients at a dose of 130 mg/m2 on day 1. Patients received either S-1 (40 mg/m2) or capecitabine (1,000 mg/m2), twice a day for 2 weeks, followed by a 1-week rest.

Results: Forty-two patients were assigned to the OS arm and 44 to the XELOX arm. The overall response rate was 33.3% (95% CI, 18.8-47.2) in the OS arm and 40.9% (95% CI, 25.5-54.4) in the XELOX arm (P = 0.230). The disease control rate was significantly higher in the OS arm than the XELOX arm [92.9% (95% CI, 83.7-100) versus 77.3% (95% CI, 64.5-89.4), P = 0.044]. With a median follow up of 17.9 months, the median progression-free survival was 6.1 months in the OS arm and 7.4 months in the XELOX arm, respectively (P = 0. 599). The median survival time was 18.7 months in the OS arm and 20.1 months in the XELOX arm (P = 0.340). The most common grade 3/4 hematologic toxicity was thrombocytopenia in both arms (19.0% for OS and 28.6% for XELOX). Grade 3/4 neutropenia was observed more frequently in the XELOX arm than the OS arm (16.7% vs. 2.4%, P = 0.026).

Conclusion: Both OS and XELOX were effective and well tolerated in patients with metastatic CRC. Our results indicate that the combination of oxaliplatin and S-1 is a possible additional therapeutic strategy for such patients.

Tuesday, September 13, 2016 8:19 PM|Xiaoqun Lv, Lingyun Zhang, Yanyan Zhu, Harun M. Said, Jimin Shi, Guoxiong Xu|Journal of Cancer (RSS 2.0)|Labels: CRC

Colorectal cancer (CRC) is the third most common cancer disease. Here we examined Nampt expression in patients with CRC and the effect of Nampt on cell viability in CRC cells. Nampt protein was overexpressed in colorectal adenoma as well as colorectal carcinoma. The immunoreactive staining of Nampt was negative in the adjacent normal colorectal tissue, weak in colorectal adenoma, and strong in colorectal carcinoma, which may represent tumor progression. Further evaluation of clinical data showed that Nampt expression was not correlated with the clinicopathological characteristics of CRC. Additionally, our in vitro studies demonstrated that Nampt promotes CRC cell viability, whereas the Nampt inhibitor FK866 suppressed CRC cell viability, which was in concordance with the previous studies in other cancer cells. Treatment with Nampt-siRNA reduced the Nampt protein expression resulting in the inhibition of the cell viability of HCT116 and Caco2. Thus, the involvement of Nampt in cell growth indicates that Nampt may play an important role in colorectal tumorigenesis. As a consequence, our results suggest that Nampt may be considered as a progression marker of colorectal tumor and a potentially therapeutic target for the treatment of CRC.

Tuesday, September 13, 2016 8:19 PM|Ana Barat, Heather J. Ruskin|Journal of Cancer (RSS 2.0)|Labels: CRC, biomarker diagnostic

Abnormal DNA-methylation is well known to play an important role in cancer onset and development, and colon cancer is no exception to this rule. Recent years have seen the increased use of large-scale technologies, (such as methylation microarray assays or specific sequencing of methylated DNA), to determine whole genome profiles of CpG island methylation in tissue samples. Comprehensive study of methylation array data from transcriptome high-throughput platforms permits determination of gene methylation markers, important for cancer profiling. Here, three large-scale methylation datasets for colon cancer have been compared to determine locus-specific methylation agreement. These data are from the GEO database, where colon cancer and apparently healthy adjacent tissues are represented by sample sizes 125 and 29 respectively in the first dataset, 24 of each in the second and 118 of each in the third. Several data analysis techniques have been employed, including Clustering, Discriminant Principal Component Analysis, Discriminant Analysis and ROC curves, in order (i) to obtain a better insight on the locus-specific concomitant methylation structures for these diverse data and (ii) to determine a robust potential marker set for indicative screening, drawn from all data taken together. The extent of the agreement between the analysed datasets is reported. Further, potential screening methylation markers, for which methylation profiles are consistent across tissue samples and several datasets, are highlighted and discussed.

Tuesday, September 13, 2016 8:19 PM|Wei-Wei Xiao, Lu-Ning Zhang, Kai-Yun You, Rong Huang, Xin Yu, Pei-Rong Ding, Yuan-Hong Gao|Journal of Cancer (RSS 2.0)|Labels: CEA, CRC

Neoadjuvant radio-chemotherapy followed by total mesorectal excision (TME) is the standard treatment option for stage II and III rectal cancer. However, for pT3N0 rectal cancer patients who receive upfront TME, the lack of an efficient method to predict their prognosis hampers postoperative treatment. A low lymphocyte-to-monocyte ratio (LMR) is associated with an unfavorable prognosis for certain malignancies; however, this association has not been investigated in rectal cancer. The purpose of this study was to evaluate whether LMR can predict the prognosis of pT3N0 rectal cancer patients following TME. Rectal cancer patients who received radical TME without preoperative treatment between June 2004 and Nov. 2011 at the Sun Yat-sen University Cancer Center were retrospectively reviewed. Counts for pre-surgery peripheral absolute lymphocytes and monocytes were obtained and used to calculate the LMR. A retrospective cohort of 280 pT3N0 rectal cancer patients who received TME was recruited. Significantly worse disease-free survival can be observed in patients with lower LMR levels (<3.78) using univariate and multivariate analyses (P=0.01 and P=0.015, respectively). Subgroup analysis in patients with elevated carcinoembryonic antigen (CEA) and LMR <3.78 exhibited an accumulated 5-year disease failure rate of approximately 40%, whereas patients with normal CEA regardless of LMR and patients with LMR ≥3.78 exhibited accumulated 5-year disease failure rates of only approximately 15%. Low pre-surgery peripheral LMR was significantly unfavorable for pT3N0 rectal cancer patient prognosis, especially in patients with elevated CEA. This easily obtained variable might serve as a valuable marker to predict the outcomes of pT3N0 rectal cancer and indicate appropriate postoperative management.

Tuesday, September 13, 2016 8:19 PM|Jung Han Kim, Seon Jeong Min, Hyun Joo Jang, Ji Woong Cho, Soo Ho Kim, Hyeong Su Kim|Journal of Cancer (RSS 2.0)|Labels: CRC

Background: We conducted this pooled analysis to investigate the impact of RECIST 1.1 on the selection of target lesions and classification of tumor response, in comparison with RECIST 1.0.

Methods: We searched MEDLINE and EMBASE for articles with terms of RECIST 1.0 or RECIST 1.1. We looked into all abstracts and virtual meeting presentations from the conferences of ASCO and ESMO between 2009 and 2013.

Results: There were six articles in the literature comparing the clinical impacts of RECIST 1.0 and RECIST 1.1 in patients with metastatic cancer. A total of 359 patients were recruited from the six trials; 217 with non-small cell lung cancer, 61 with gastric cancer, 58 with colorectal cancer, and 23 with thyroid cancer. The number of target lesions by RECIST 1.1 was significantly lower than that by RECIST 1.0 (P<0.001). Because of new lymph node criteria, fourteen patients (3.1%) had no target lesions when adopting RECIST 1.1. RECIST 1.1 showed high concordance with RECIST 1.0 in the assessment of tumor responses (k = 0.903). Sixteen patients (4.8%) showed disagreement between the two criteria.

Conclusion: This pooled study demonstrated that RECIST 1.1 showed a highly concordant response assessment with RECIST 1.0 in patients with metastatic cancer.

Tuesday, September 13, 2016 8:19 PM|Yingjun Quan, Ming Xu, Peng Cui, Min Ye, Biao Zhuang, Zhijun Min|Journal of Cancer (RSS 2.0)|Labels: CRC

GRHL2 was implicated in regulating cancer development. Our previous study demonstrated that knockdown GRHL2 in colorectal cancer (CRC) cells inhibited cell proliferation by targeting ZEB1. It is unclear whether GRHL2 expression may have diagnostic or prognostic value in colorectal carcinoma. Additionally, how GRHL2 is associated with the clinical features of colorectal carcinoma is not known. In current study, immunohistochemistry stains were performed to examine GRHL2 in 171 colorectal cancers and paired normal colon mucosa. The prognostic value of GRHL2 was investigated in a retrospective cohort study with a five-year follow-up. The effects of GRHL2 on cell growth in vitro and in vivo were explored by GRHL2 over-expressing in HT29 and SW620 CRC cells. Further, the regulation of cell cycle and proliferation proteins by GRHL2 were assessed by flow cytometry and western blot. We found that GRHL2 was over-expressed in CRC tissues, and played an important role in CRC tumorigenesis. GRHL2 expression positively correlated with tumor size and TNM stage. Kaplan-Meier analysis showed that GRHL2 was an independent prognostic factor for both overall survival and recurrence-free survival. Ectopic over-expression of GRHL2 in CRC cell line HT29 and SW620 induced an increase of cellular proliferation in vitro and promoting tumor growth in vivo. The acquisition of GRHL2 regulated cell cycle and modulates the expression of proliferation proteins p21, p27, cyclin A and cyclin D1. Together, our findings reveal GRHL2 can be used as a novel predictive biomarker and represent a potential therapeutic target against CRC.

Tuesday, September 13, 2016 8:19 PM|A. Orlandi, M. Di Salvatore, C. Bagalà, M. Basso, A. Strippoli, F. Plastino, M.A. Calegari, A. Cassano, A. Astone, C. Barone|Journal of Cancer (RSS 2.0)|Labels: RAS, CRC

Introduction: Oxaliplatin (Oxa) is widely used in metastatic colorectal cancer (mCRC), but currently there are not valid predictors of response to this drug. In the control arms both of OPUS and PRIME studies Oxa seems more active in patients with mCRC with mutated (mt) KRAS than in those with wild type (wt) KRAS. Recently we have retrospectively confirmed this suggestion, therefore we have hypothesized that the mutational status of KRAS could influence the expression of ERCC1, one of the main mechanisms of Oxa resistance.

Material and Methods: We used four cell lines of colorectal cancer: two KRAS wild type (wt) (HCT-8 and HT-29) and two KRAS mt (SW620 and SW480). We evaluated the sensitivity of these cell lines to Oxa by MTT-test as well the ERCC1 levels before and after 24 h exposure to Oxa by Real-Time PCR. We silenced KRAS in a KRAS mt cell line (SW620LV) to evaluate the impact on Oxa sensitivity and ERCC1 levels. Lastly, ERCC1 was also silenced in order to confirm the importance of this protein as an Oxa resistance factor.

Results: The KRAS mt cell lines resulted more sensitive to Oxa (OR 2.68; IC 95% 1.511-4.757 p<0.001). The basal levels of ERCC1 did not show significant differences between KRAS mt and wt cell lines, however, after 24 h exposure to Oxa, only the wt KRAS lines showed the ability to induce ERCC1, with a statistically significant difference (OR 42.9 IC 95% 17.260-106.972 p<0.0005). By silencing KRAS, sensitivity to Oxa was reduced in mt KRAS cell lines and this effect was associated with the acquisition of ability to induce ERCC1. Silencing of ERCC1, in turn, enhanced the sensitivity to Oxa in wt KRAS cell lines and restored sensitivity to Oxa in SW620LV cell line.

Conclusion: KRAS mutated cell lines were more sensitive to Oxa. This feature seems secondary to the inability of these cells to induce ERCC1 after exposure to Oxa. Thus, KRAS mutational status might be a predictor of response to Oxa in CRC surrogating the cell ability to induce ERCC1.

Tuesday, September 13, 2016 8:19 PM|Rezwan Islam, Po-Huang Chyou, James K Burmester|Journal of Cancer|Labels: VEGF, CRC

Purpose: Bevacizumab, an FDA-approved adjuvant treatment for metastatic colon cancer, has extended survival for many patients. However, factors predicting response to treatment remain undefined.

Patients and Methods: Relevant clinical and environmental data were abstracted from medical records of 149 evaluable patients treated with bevacizumab for metastatic colon cancer at a multi-specialty clinic. Tumor response was calculated from radiologic reports using Response Evaluation Criteria in Solid Tumors (RECIST) criteria and verified by oncologist review. Patients with at least one occurrence of complete or partial response or stable disease were classified as responders; those exhibiting progressive disease were classified as non-responders.

Results: Univariate analysis demonstrated that blood in stool (P<0.05), unexplained weight loss (P<0.05), primary colon cancer site (P<0.05), chemotherapy treatment of primary tumor site (P<0.05), and adenocarcinoma versus adenoma subtype (P<0.05) was associated with tumor responsiveness. Factors remaining statistically significant following multivariate modeling included adenocarcinoma as tumor cell type versus other adenocarcinoma subtypes

(OR=6.35, 95% CI: 1.08-37.18), chemotherapy treatment applied to primary tumor (OR= 0.07, 95% CI: 0.0-0.76,), tumor localization to cecal/ascending colon (OR=0.061, 95% CI: 0.006-0.588,), and unexplained weight loss (OR=0.1, 95% CI: 0.02-0.56,). Chemotherapy treatment of primary tumor, unexplained weight loss, and cecal/ascending localization of the tumor were associated with poorer outcomes. Adenocarcinoma as cell type compared to other adenocarcinoma subtypes was associated with better response to bevacizumab treatment.

Conclusion: Results suggest that response to bevacizumab therapy may be predicted by modeling clinical factors including symptomology on presentation, tumor location and type, and initial response to chemotherapy.

Tuesday, September 13, 2016 8:19 PM|Haggi Mazeh, Ido Mizrahi, Nadia Ilyayev, David Halle, Björn LDM Brücher, Anton Bilchik, Mladjan Protic, Martin Daumer, Alexander Stojadinovic, Itzhak Avital, Aviram Nissan|Journal of Cancer|Labels: CRC, clinical trial

The discovery of microRNA, a group of regulatory short RNA fragments, has added a new dimension to the diagnosis and management of neoplastic diseases. Differential expression of microRNA in a unique pattern in a wide range of tumor types enables researches to develop a microRNA-based assay for source identification of metastatic disease of unknown origin. This is just one example of many microRNA-based cancer diagnostic and prognostic assays in various phases of clinical research.

Since colorectal cancer (CRC) is a phenotypic expression of multiple molecular pathways including chromosomal instability (CIN), micro-satellite instability (MIS) and CpG islands promoter hypermethylation (CIMP), there is no one-unique pattern of microRNA expression expected in this disease and indeed, there are multiple reports published, describing different patterns of microRNA expression in CRC.

The scope of this manuscript is to provide a comprehensive review of the scientific literature describing the dysregulation of and the potential role for microRNA in the management of CRC. A Pubmed search was conducted using the following MeSH terms, "microRNA" and "colorectal cancer". Of the 493 publications screened, there were 57 papers describing dysregulation of microRNA in CRC.

Tuesday, September 13, 2016 8:19 PM|Terry Hyslop, Scott A. Waldman|Journal of Cancer|Labels: CRC

Metastatic disease is the principle cause of death from colorectal cancer. In that context, the most significant indicator of overall survival and therapeutic response to adjuvant chemotherapy is the presence of metastatic tumor cells in regional lymph nodes. Although histopathologic analysis of lymph nodes is central to all colorectal cancer staging paradigms, its prognostic and predictive value is limited. Indeed, about 30% of patients with histopathology-negative lymph nodes (pN0) die from metastatic disease, reflected by microscopic lymph node metastases that are overlooked by standard techniques. These unrecognized tumor cells are especially important when considering racial disparities in outcomes in colorectal cancer patients, where blacks with lymph node-negative disease have the largest discrepancies in outcomes, with more than 40% excess mortality compared to Caucasian patients. However, the significance of tumor cells in regional lymph nodes remains uncertain, and approximately 50% of colorectal cancer patients with nodal metastases detected by histopathology remain free of recurrent disease. Accurate identification of occult metastases in regional lymph nodes, and defining their value as prognostic markers of recurrence risk and predictive markers of response to adjuvant chemotherapy remains one challenge in the management of colorectal cancer patients. Guanylyl cyclase C (GUCY2C), a receptor which is expressed primarily in intestinal cells normally, but is universally over-expressed by colorectal cancer cells, has been validated to detect prognostically significant occult metastases using quantitative RT-PCR (RT-qPCR). Biomarker validation was achieved through a prospective, multicenter, blinded clinical trial. In that trial, occult tumor burden estimated across all regional lymph nodes by GUCY2C RT-qPCR predicted clinical outcomes, identifying node-negative patients with a low (near zero) risk, and those with >80% risk, of developing disease recurrence. Moreover, there was disproportionately higher occult tumor burden in black, compared to white, patients which contributes to racial disparities in outcomes in colorectal cancer. The diagnostic paradigm quantifying occult tumor burden using GUCY2C qRT-PCR is positioned to reduce racial disparities in colorectal cancer mortality.

Tuesday, September 13, 2016 8:19 PM|Michael Bordonaro|Journal of Cancer|Labels: WNT, CRC

RNA processing involves a variety of processes affecting gene expression, including the removal of introns through RNA splicing, as well as 3' end processing (cleavage and polyadenylation). Alternative RNA processing is fundamentally important for gene regulation, and aberrant processing is associated with the initiation and progression of cancer. Deregulated Wnt signaling, which is the initiating event in the development of most cases of human colorectal cancer (CRC), has been linked to modified RNA processing, which may contribute to Wnt-mediated colonic carcinogenesis. Crosstalk between Wnt signaling and alternative RNA splicing with relevance to CRC includes effects on the expression of Rac1b, an alternatively spliced gene associated with tumorigenesis, which exhibits alternative RNA splicing that is influenced by Wnt activity. In addition, Tcf4, a crucial component of Wnt signaling, also exhibits alternative splicing, which is likely involved in colonic tumorigenesis. Modulation of 3' end formation, including of the Wnt target gene COX-2, also can influence the neoplastic process, with implications for CRC. While many human genes are dependent on introns and splicing for normal levels of gene expression, naturally intronless genes exist with a unique metabolism that allows for intron-independent gene expression. Effects of Wnt activity on the RNA metabolism of the intronless Wnt-target gene c-jun is a likely contributor to cancer development. Further, butyrate, a breakdown product of dietary fiber and a histone deacetylase inhibitor, upregulates Wnt activity in CRC cells, and also modulates RNA processing; therefore, the interplay between Wnt activity, the modulation of this activity by butyrate, and differential RNA metabolism in colonic cells can significantly influence tumorigenesis. Determining the role played by altered RNA processing in Wnt-mediated neoplasia may lead to novel interventions aimed at restoring normal RNA metabolism for therapeutic benefit. Therefore, this minireview presents a brief overview of several aspects of RNA processing of relevance to cancer, which potentially influence, or are influenced by, Wnt signaling activity.

Tuesday, September 13, 2016 8:19 PM|Darina L Lazarova, Terrence Wong, Christopher Chiaro, Eric Drago, Michael Bordonaro|Journal of Cancer|Labels: WNT, CRC

Deregulated WNT/catenin pathway, usually resulting from mutations in the adenomatous polyposis coli and beta-catenin genes, drives colorectal tumorigenesis. Dietary fiber has been shown to have a protective role against colorectal cancer (CRC). We have previously demonstrated that the histone deacetylase inhibitor (HDACi) butyrate, a fermentation product of dietary fiber, induces WNT/catenin hyperactivation, which promotes CRC cell apoptosis. Therefore, the ability of butyrate to induce WNT hyperactivation and thus promote CRC cell apoptosis may in part explain the preventive function of fiber against CRC. The association between beta-catenin and the transcriptional coactivator p300 may influence WNT/catenin signaling and, therefore, colonic cell physiology. p300 functions as a histone acetylase (HAT); therefore, the modulation of WNT/catenin activity by p300 may influence the ability of the HDACi butyrate to hyperinduce WNT signaling and apoptosis in CRC cells. Our findings indicate that p300 affects the hyperinduction of WNT activity by butyrate. Knockdown of p300 levels represses butyrate-mediated WNT/catenin activity; but still allows for butyrate-mediated apoptosis. Overexpression of p300 stimulates basal and butyrate-induced WNT signaling in some, but not all, CRC cell lines. We also evaluate the role of p300 in therapeutic approaches that target CBP. The small molecule ICG-001, in clinical trial, is a specific inhibitor of CBP-mediated WNT signaling, and previous studies have suggested that p300 is required for the activity of ICG-001. However, we report that ICG-001 maintains full activity against CBP-mediated WNT signaling in p300-deficient cell lines, including the butyrate-resistance line HCT-R. In addition, our findings evaluating combinatorial treatment of ICG-001 and butyrate in HCT-R cells may have important therapeutic implications for the treatment of butyrate-resistant CRCs.

Tuesday, September 13, 2016 8:19 PM|Darina L Lazarova, Christopher Chiaro, Terrence Wong, Eric Drago, Anthony Rainey, Shannon O'Malley, Michael Bordonaro|Journal of Cancer|Labels: HDAC, WNT, CRC

Mutations in the WNT/beta-catenin pathway are responsible for initiating the majority of colorectal cancers (CRCs). We have previously shown that hyperactivation of this signaling by histone deacetylase inhibitors (HDACis) such as butyrate, a fermentation product of dietary fiber, promotes CRC cell apoptosis. The extent of association between beta-catenin and the transcriptional coactivator CREB-binding protein (CBP) influences WNT/catenin signaling and, therefore, colonic cell physiology. CBP functions as a histone acetylase (HAT); therefore, we hypothesized that the modulation of WNT/catenin activity by CBP modifies the ability of the HDACi butyrate to hyperinduce WNT signaling and apoptosis in CRC cells. Our findings indicate that CBP affects the hyperinduction of WNT activity by butyrate. ICG-001, which specifically blocks association between CBP and beta-catenin, abrogates the butyrate-triggered increase in the number of CRC cells with high levels of WNT/catenin signaling. Combination treatment of CRC cells with ICG-001 and butyrate results in cell type-specific effects on apoptosis. Further, both butyrate and ICG-001 repress CRC cell proliferation, with additive effects in suppressing cell growth. Our study strongly suggests that ICG-001-like agents would be effective against butyrate/HDACi-resistant CRC cells. Therefore, ICG-001-like agents may represent an important therapeutic option for CRCs that exhibit low-fold hyperactivation of WNT activity and apoptosis in the presence of HDACis. The findings generated from this study may lead to approaches that utilize modulation of CBP activity to facilitate CRC therapeutic or chemopreventive strategies.

Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Dev, K.; Veerendrakumar, K.; Krishnamurthy, S.;...
Background : The en bloc excision of primary tumor along with locoregional lymphadenectomy is the standard curative surgical treatment for rectal cancer. The development of local or locoregional recurrence is primarily due to sub-optimal surgical ...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: RAS, CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Passot, G.; Chun, Y.S.; Kopetz, S.E.;...
Introduction For patients with stage IV colorectal cancer, the liver is the most common site of distant metastases. Advances in modern chemotherapy and targeted agents have significantly improved response and survival rates for patients with color...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Braam, H.J.W.; Hoogwater, F.J.H.; Van Oudheusden, T.R.;...
Background : The aim of the current study is to evaluate our experience in colorectal cancer patients with synchronous peritoneal and liver metastases treated with cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC). ...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Gervaz, P.; Usel, M.; Rapiti, E.;...
Introduction Clinical outcome of colon cancer (CCacrnm1) is continuously improving in Europe, North America, and Asia. Implementation of screening programmes, facilitated access to colonoscopy, and development of efficient chemotherapy regimen, ar...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Van der Valk, M.; Hilling, D.; Meershoek-Klein Kranenbarg, E.;...
Background : In 2014 the International Watch-and-Wait Database (IWWD) for rectal cancer was established under the umbrella of EURECCA and the Champalimaud Foundation. The main goal of this database is to collect all available retrospective and pro...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Zoheir, M.
Background : The mucus layer coating the gastrointestinal tract is the front line of innate host defense. MUC2 is the major secretory mucin synthesized and secreted by goblet cells, whereas goblet and absorptive cells express membrane-bound mucins...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Baek, M.J.; Park, D.K.; Lee, S.J.;...
Background : Lymphocyte antigen 6 complex locus (LY6Eacrnm1) is a glycosylphosphatidylinositol (GPI)-linked cell-surface protein that is induced by Interferon (IFN). * Lymphocyte antigen 6 complex locus * mRNA was found to be overexpressed in colo...
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Forlin, M.; Olivieri, M.; Bertola, G.;...
Background : Preoperative chemoradiation therapy (CRTacrnm1) followed by radical surgery is standard of care in patients with locally advanced mid-distal rectal cancer (LARC). Pathological complete response in the primary tumor (ypCR) is observed ...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC, biomarker diagnostic
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Bobowicz, M.; Skrzypski, M.; Czapiewski, P.;...
Background : The role of adjuvant chemotherapy in stage T2-T3N0 colon cancer (CCacrnm1) is controversial and there are currently no reliable factors allowing for individual selection of patients with high risk of relapse. We searched for microRNA-...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Park, D.G.; Namgung, W.; Yun, J.S.;...
Background : While colorectal cancer is gradually getting better treatment results in the development of various treatment methods, metastasis occurs in about 30% and that eventually leading to death. Difference of the genetic characteristics betw...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Zawadzki, M.; Rzaca, M.; Czarnecki, R.;...
Background : Adoption of robotic technology in the field of colorectal surgery is slow but continues to increase. The advantages of robotics over laparoscopy involve lower rates of conversion and a steeper learning curve. Typically, what is publis...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Brzuszkiewicz, K.; Pietruszka, S.; Paszko, A.;...
Chemokines are the chemotactic cytokines. They play an important role in induction and maintenance of immune reactions by regulation of migration of immunocompetent cells, angiogenesis and migration of stem cells. They have also been shown to regu...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC, biomarker diagnostic
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Lee, S.J.; Baek, M.J.; Park, D.K.;...
Background : Solute Carrier Organic Anion Transporter Family Member 4A1 (SLCO4A1acrnm1) is involved in glucose, bile salts and organic acids, metal ions and amine compounds transport. Many reseachers reported that * Solute Carrier Organic Anion Tr...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Um, J.W.; Baek, M.J.
Background : PRSS33, one of serine protease multigene family, has central roles in the regulation of a wide variety of physiological processes, including inflammation, development and malignancy. However, the function of this gene in colorectal ca...
Tuesday, September 13, 2016 4:20 PM|ClinicalKey Latest Issue: European Journal of Surgical Oncology|Labels: CRC
European Journal of Surgical Oncology, Volume 42, Issue 9, 2016 Sep | Neofytou, K.; Petrou, A.; Petrides, C.;...
Background : Loss of Stromal Caveolin-1 (CAV1acrnm1) expression is associated with poor prognosis in various cancers. In this study, we evaluated the prognostic value of * Caveolin 1 * expression of both tumour cells and stromal cells in colorecta...
Tuesday, September 13, 2016 2:05 PM|Paul Zarogoulidis, Sofia Lampaki, Lonny Yarmus, Ioannis Kioumis, Georgia Pitsiou, Nikolaos Katsikogiannis, Wolfgang Hohenforst-Schmidt, Qiang Li, Haidong Huang, Antonios Sakkas, John Organtzis, Leonidas Sakkas, Ioannis Mpoukovinas, Kosmas Tsakiridis, George Lazaridis, Konstantinos Syrigos, Konstantinos Zarogoulidis|Journal of Cancer|Labels: CRC, IL

Interleukin 7 and 15 are considered powerful pro-inflammatory cytokines, they have the ability to destabilize chromosomes and induce tumorigenesis. Additionally, they can control malignancy proliferation by influencing the tumor microenvironment and immune system. Immunotherapy has been proposed as a treatment modality for malignancy for over a decade; the exact mechanisms of action and pathways are still under investigation. Interleukin 7 and 15 have been extensively investigated in hematological malignancies since their mode of action influences the stimulation of the immune system in a more direct way than other malignancies such as lung, melanoma, and breast, renal and colorectal cancer.

Tuesday, September 13, 2016 2:05 PM|Aziz Zaanan, Cécile Dalban, Jean-François Emile, Hélène Blons, Jean-François Fléjou, Claire Goumard, Melek Istanbullu, Claire Calmel, Khalid Alhazmi, Pierre Validire, Christophe Louvet, Aimery de Gramont, Pierre Laurent-Puig, Julien Taïeb, Françoise Praz|Journal of Cancer|Labels: CRC

Background: While single nucleotide polymorphisms (SNP) in genes involved in DNA repair or drug metabolism have been shown to influence survival of metastatic colon cancer patients treated with FOLFOX, data on adjuvant setting are scarce.

Methods: This study evaluated the correlation between disease-free survival (DFS) of 210 unselected stage III colon cancer patients receiving FOLFOX chemotherapy, and ERCC1-118 (rs11615, c.354T>C), XRCC1-399 (rs25487, c.1196G>A) and GSTP1-105 (rs1695, c.313A>G) polymorphisms. SNP were determined on tumor DNA using a PCR-based RFLP technique.

Results: In univariate analysis, a trend towards longer DFS was observed for ERCC1 (C/T + T/T) versus (C/C) (HR=2.29; p=0.06), and XRCC1 (A/A) versus (G/G + G/A) (HR=1.61; p=0.16), but not for GSTP1 genotypes; a statistically significant p value was obtained when combining ERCC1 and XRCC1 favorable genotypes (0 versus ≥ 1 favorable genotypes, HR=2.42; p=0.02). After adjustment on tumor stage, lymph node ratio and differentiation grade, multivariate analysis showed that combining ERCC1 and XRCC1 genotypes gave a p value slightly above the threshold for statistical significance (HR=2.03; p=0.06), which was lower than for tumor stage, lymph node ratio or differentiation grade.

Conclusion: The association of ERCC1 and XRCC1 polymorphisms may influence the prognosis of stage III colon cancer patients treated with FOLFOX adjuvant chemotherapy. Yet, these findings need to be confirmed in independent prospective studies.

Tuesday, September 13, 2016 2:05 PM|Avery S. Walker, Eric K. Johnson, Justin A. Maykel, Alex Stojadinovic, Aviram Nissan, Bjorn Brucher, Bradley J. Champagne, Scott R. Steele|Journal of Cancer|Labels: CRC, clinical trial

Surgical resection remains a mainstay of treatment and is highly effective for localized colorectal cancer. However, ~30-40% of patients develop recurrence following surgery and 40-50% of recurrences are apparent within the first few years after initial surgical resection. Several variables factor into the ultimate outcome of these patients, including the extent of disease, tumor biology, and patient co-morbidities. Additionally, the time from initial treatment to the development of recurrence is strongly associated with overall survival, particularly in patients who recur within one year of their surgical resection. Current post-resection surveillance strategies involve physical examination, laboratory, endoscopic and imaging studies utilizing various high and low-intensity protocols. Ultimately, the goal is to detect recurrence as early as possible, and ideally in the asymptomatic localized phase, to allow initiation of treatment that may still result in cure. While current strategies have been effective, several efforts are evolving to improve our ability to identify recurrent disease at its earliest phase. Our aim with this article is to briefly review the options available and, more importantly, examine emerging and future options to assist in the early detection of colon and rectal cancer recurrence.

Tuesday, September 13, 2016 2:05 PM|Avery S. Walker, Nathan P. Zwintscher, Eric K. Johnson, Justin A. Maykel, Alexander Stojadinovic, Aviram Nissan, Itzhak Avital, Björn LDM Brücher, Scott R. Steele|Journal of Cancer|Labels: CRC

Treatment of advanced colon and rectal cancer has significantly evolved with the introduction of neoadjuvant chemoradiation therapy so much that, along with more effective chemotherapy regimens, surgery has been considered unnecessary among some institutions for select patients. The tumor response to these treatments has also improved and ultimately has been shown to have a direct effect on prognosis. Yet, the best way to monitor that response, whether clinically, radiologically, or with laboratory findings, remains controversial. The authors' aim is to briefly review the options available and, more importantly, examine emerging and future options to assist in monitoring treatment response in cases of locally advanced rectal cancer and metastatic colon cancer.

Monday, September 12, 2016 1:07 PM|Benson, A. B., Kiss, I., Bridgewater, J., Eskens, F. A. L. M., Sasse, C., Vossen, S., Chen, J., Van Sant, C., Ball, H. A., Keating, A., Krivoshik, A.|Clinical Cancer Research Online First Articles|Labels: VEGF, CRC, clinical trial

Purpose: Tivozanib, a selective inhibitor of VEGFR-1, -2, and -3, plus mFOLFOX6 in an advanced gastrointestinal cancer phase Ib study had encouraging antineoplastic activity and a tolerable safety profile. This randomized, open-label, phase II trial of tivozanib/mFOLFOX6 versus bevacizumab/mFOLFOX6 in patients with previously untreated metastatic colorectal cancer (mCRC) evaluated tivozanib activity versus bevacizumab.

Experimental Design: Treatment-naïve patients received mFOLFOX6 every 2 weeks of each 28-day cycle plus either tivozanib orally 1.5 mg once daily for 21 days or bevacizumab intravenously 5 mg/kg every 2 weeks. Investigator-assessed progression-free survival (PFS) was the primary endpoint; some secondary endpoints included safety, overall survival, overall response rate (ORR), duration of response, time to treatment failure, and biomarker subgroup analyses.

Results: A prespecified interim futility analysis demonstrated that the futility boundary for superiority of tivozanib/mFOLFOX6 over bevacizumab/mFOLFOX6 for PFS in the intent-to-treat population was crossed; median PFS was 9.4 versus 10.7 months [HR = 1.091; confidence interval (CI), 0.693–1.718; P = 0.706]. Tivozanib/mFOLFOX6 resulted in PFS and ORR comparable with bevacizumab/mFOLFOX6; interim analyses biomarker results revealed no significant PFS association. Post hoc final analyses demonstrated a potential difference in tivozanib-specific PFS in patients with low neuropilin-1 (NRP-1), but not in patients with high NRP-1. Tivozanib/mFOLFOX6 was tolerable and adverse events were comparable with both bevacizumab/mFOLFOX6 and previous tivozanib studies.

Conclusions: The efficacy of tivozanib/mFOLFOX6 was comparable with but not superior to bevacizumab/mFOLFOX6 in patients with previously untreated mCRC. Since data from the prespecified interim analysis did not demonstrate superiority, this resulted in discontinuation of the study. The safety and tolerability profile of tivozanib/mFOLFOX6 was consistent with other tivozanib trials. NRP-1 is a potential predictive biomarker for tivozanib activity, but these results require further validation. Clin Cancer Res; 1–10. ©2016 AACR.

Sunday, September 11, 2016 11:00 PM|Guo, Xiaoqing; Meng, Yue; Sheng, Xiaotong; Guan, Yuan; Zhang, Fenglei; Han, Zhen; Kang, Yuying; Tai, Guihua; Zhou, Yifa; Cheng, Hairong|Anti-Cancer Drugs - Published Ahead-of-Print|Labels: EGFR, Jnk, TNF, CRC
Tumor necrosis factor related apoptosis-inducing ligand (TRAIL) is a cytokine that selectively induces apoptosis in many tumor cells while leaving normal cells intact and is thus an attractive candidate for antitumor therapies. This paper reports that the combination of tunicamycin plus TRAIL produced a strong synergistic effect in TRAIL-sensitive human colon cancer HCT116 cells and TRAIL-resistant HT-29 cells. On a cellular mechanistic level, tunicamycin-enhanced TRAIL-induced apoptosis by death receptor (DR) 5 upregulation and DR4 deglycosylation. Knockdown of DR5 but not DR4 expression by specific shRNAs or siRNAs significantly increased tunicamycin-mediated and TRAIL-mediated cell viability. DR5 induction was regulated by C/EBP homologous protein (CHOP) and JNK as CHOP siRNA or JNK inhibitor SP600125 considerably abolished the DR5 induction. In addition, tunicamycin inhibited epidermal growth factor receptor glycosylation and the downstream signaling pathways, Akt and extracellular signal-regulated kinases activation, which might also be required for TRAIL sensitization by tunicamycin. In summary, tunicamycin effectively enhanced TRAIL-induced apoptosis might through JNK-CHOP-mediated DR5 upregulation and the inhibition of the epidermal growth factor receptor pathway. Copyright (C) 2016 Wolters Kluwer Health, Inc. All rights reserved.
Wednesday, September 7, 2016 8:45 AM|Ciarloni, L., Ehrensberger, S. H., Imaizumi, N., Monnier-Benoit, S., Nichita, C., Myung, S.-J., Kim, J. S., Song, S. Y., Kim, T. I., van der Weg, B., Meier, R., Borovicka, J., Beglinger, C., Vallet, C., Maerten, P., Rüegg, C., Dorta, G.|Clinical Cancer Research Online First Articles|Labels: CEA, CRC

Purpose: A blood test for early detection of colorectal cancer is a valuable tool for testing asymptomatic individuals and reducing colorectal cancer–related mortality. The objective of this study was to develop and validate a novel blood test able to differentiate patients with colorectal cancer and adenomatous polyps (AP) from individuals with a negative colonoscopy.

Experimental Design: A case–control, multicenter clinical study was designed to collect blood samples from patients referred for colonoscopy or surgery. Predictive algorithms were developed on 75 controls, 61 large AP (LAP) ≥1 cm, and 45 colorectal cancer cases and independently validated on 74 controls, 42 LAP, and 52 colorectal cancer cases (23 stages I–II) as well as on 245 cases including other colorectal findings and diseases other than colorectal cancer. The test is based on a 29-gene panel expressed in peripheral blood mononuclear cells alone or in combination with established plasma tumor markers.

Results: The 29-gene algorithm detected colorectal cancer and LAP with a sensitivity of 79.5% and 55.4%, respectively, with 90.0% specificity. Combination with the protein tumor markers carcinoembryonic antigen (CEA) and CYFRA21-2 resulted in a specificity increase (92.2%) with a sensitivity for colorectal cancer and LAP detection of 78.1% and 52.3%, respectively.

Conclusions: We report the validation of a novel blood test, Colox®, for the detection of colorectal cancer and LAP based on a 29-gene panel and the CEA and CYFRA21-1 plasma biomarkers. The performance and convenience of this routine blood test provide physicians a useful tool to test average-risk individuals unwilling to undergo upfront colonoscopy. Clin Cancer Res; 22(18); 1–8. ©2016 AACR.

Wednesday, September 7, 2016 8:45 AM|Bosch, L. J. W., Luo, Y., Lao, V. V., Snaebjornsson, P., Trooskens, G., Vlassenbroeck, I., Mongera, S., Tang, W., Welcsh, P., Herman, J. G., Koopman, M., Nagtegaal, I. D., Punt, C. J. A., van Criekinge, W., Meijer, G. A., Monnat, R. J., Carvalho, B., Grady, W. M.|Clinical Cancer Research Online First Articles|Labels: CRC, clinical trial

Purpose: WRN promoter CpG island hypermethylation in colorectal cancer has been reported to increase sensitivity to irinotecan-based therapies. We aimed to characterize methylation of the WRN promoter, determine the effect of WRN promoter hypermethylation upon expression, and validate a previous report that WRN promoter hypermethylation predicts improved outcomes for patients with metastatic colorectal cancer (mCRC) treated with irinotecan-based therapy.

Experimental Design: WRN methylation status was assessed using methylation-specific PCR and bisulfite sequencing assays. WRN expression was determined using qRT-PCR and Western blotting. WRN methylation status was correlated with overall survival (OS) and progression-free survival (PFS) in 183 patients with mCRC. Among these patients, 90 received capecitabine monotherapy as first-line therapy, and 93 received capecitabine plus irinotecan (CAPIRI) therapy as part of the CAIRO phase III clinical trial.

Results: WRN mRNA and WRN protein expression levels were low in colorectal cancer cell lines and in primary colorectal cancer and were largely independent of WRN methylation status. Patients with methylated WRN colorectal cancer had a shorter OS compared with patients who had unmethylated WRN colorectal cancer (HR = 1.6; 95% confidence interval [CI], 1.2–2.2; P = 0.003). Patients with unmethylated WRN showed a significantly longer PFS when treated with CAPIRI compared with capecitabine alone (HR = 0.48; 95% CI, 0.32–0.70; P = 0.0001). In contrast, patients did not benefit from adding irinotecan to capecitabine when WRN was methylated (HR = 1.1; 95% CI, 0.69–1.77; P = 0.7).

Conclusions: WRN expression is largely independent of WRN promoter hypermethylation in colorectal cancer. Moreover, we could not validate the previous finding that WRN promoter hypermethylation predicts improved clinical outcomes of mCRC treated with irinotecan-based therapy and found instead the opposite result. Clin Cancer Res; 22(18); 1–11. ©2016 AACR.

Sunday, September 4, 2016 6:00 PM|Giuseppe Toffoli|International Journal of Molecular Sciences|Labels: CRC, biomarker diagnostic
Background: Pathological complete response (pCR) to neoadjuvant chemoradiotherapy (CRT) in locally advanced rectal cancer (LARC) is still ascribed to a minority of patients. A pathway based-approach could highlight the predictive role of germline single nucleotide polymorphisms (SNPs). The primary aim of this study was to define new predictive biomarkers considering treatment specificities. Secondary aim was to determine new potential predictive biomarkers independent from radiotherapy (RT) dosage and cotreatment with oxaliplatin. Methods: Thirty germ-line SNPs in twenty-one genes were selected according to a pathway-based approach. Genetic analyses were performed on 280 LARC patients who underwent fluoropyrimidine-based CRT. The potential predictive role of these SNPs in determining pathological tumor response was tested in Group 1 (94 patients undergoing also oxaliplatin), Group 2 (73 patients treated with high RT dosage), Group 3 (113 patients treated with standard RT dosage), and in the pooled population (280 patients). Results: Nine new predictive biomarkers were identified in the three groups. The most promising one was rs3136228-MSH6 (p = 0.004) arising from Group 3. In the pooled population, rs1801133-MTHFR showed only a trend (p = 0.073). Conclusion: This exploratory study highlighted new potential predictive biomarkers of neoadjuvant CRT and underlined the importance to strictly define treatment peculiarities in pharmacogenetic analyses.
Friday, September 2, 2016 8:31 AM|Onclive Colorectal Cancer Articles|Labels: CRC
Molecular testing for colorectal cancer continues to evolve at a rapid pace. Biomarkers already have the potential to predict outcomes and better target therapies, with the eventual goal to completely personalize treatments based on the patient’s individual biomarker profile and other tumor markers. 
Thursday, September 1, 2016 10:05 PM|Bertino, E. M., McMichael, E. L., Mo, X., Trikha, P., Davis, M., Paul, B., Grever, M., Carson, W. E., Otterson, G. A.|Molecular Cancer Therapeutics current issue|Labels: CRC, HNN, clinical trial

mAbs can induce antibody-dependent cellular cytotoxicity (ADCC) via the innate immune system's ability to recognize mAb-coated cancer cells and activate immune effector cells. Lenalidomide is an immunomodulatory agent with the capacity to stimulate immune cell cytokine production and ADCC activity. This phase I trial evaluated the combination of cetuximab with lenalidomide for the treatment of advanced colorectal and head and neck squamous cell cancers (HNSCC). This trial included patients with advanced colorectal cancer or HNSCC. Treatment consisted of cetuximab 500 mg/m2 i.v. every two weeks with lenalidomide given orally days 1–21 on a 28-day cycle. Three dose levels of lenalidomide were evaluated (15, 20, 25 mg). Correlative studies included measurement of ADCC, FcRIIIA polymorphism genotyping, measurement of serum cytokine levels, and flow cytometric analysis of immune cell subtypes. Twenty-two patients were enrolled (19 colorectal cancer, 3 HNSCC). Fatigue was the only dose-limiting toxicity. One partial response was observed and 8 patients had stable disease at least 12 weeks. The recommended phase II dose is cetuximab 500 mg/m2 with lenalidomide 25 mg daily, days 1–21. Correlative studies demonstrated a dose-dependent increase in natural killer cytotoxic activity with increasing doses of lenalidomide. Cetuximab and lenalidomide were well tolerated. There was a lenalidomide dose-dependent increase in ADCC with higher activity in patients enrolled in cohort 3 than those enrolled in cohorts 1/2. Although response was not a primary endpoint, there was evidence of antitumor activity for the combination therapy. Further investigation of lenalidomide as an immunomodulator in solid tumors is warranted. Mol Cancer Ther; 15(9); 2244–50. ©2016 AACR.

Thursday, September 1, 2016 10:05 PM|Fan, Y.-Y., Callaway, E., M. Monk, J., S. Goldsby, J., Yang, P., Vincent, L., S. Chapkin, R.|Cancer Prevention Research recent issues|Labels: CRC

A significant increase in cyclooxygenase 2 (COX2) gene expression has been shown to promote cylcooxygenase-dependent colon cancer development. Controversy associated with the role of COX2 inhibitors indicates that additional work is needed to elucidate the effects of arachidonic acid (AA)-derived (cyclooxygenase and lipoxygenase) eicosanoids in cancer initiation, progression, and metastasis. We have recently developed a novel Fads1 knockout mouse model that allows for the investigation of AA-dependent eicosanoid deficiency without the complication of essential fatty acid deficiency. Interestingly, the survival rate of Fads1-null mice is severely compromised after 2 months on a semi-purified AA-free diet, which precludes long-term chemoprevention studies. Therefore, in this study, dietary AA levels were titrated to determine the minimal level required for survival, while maintaining a distinct AA-deficient phenotype. Null mice supplemented with AA (0.1%, 0.4%, 0.6%, 2.0%, w/w) in the diet exhibited a dose-dependent increase (P < 0.05) in AA, PGE2, 6-keto PGF, TXB2, and EdU-positive proliferative cells in the colon. In subsequent experiments, null mice supplemented with 0.6% AA diet were injected with a colon-specific carcinogen (azoxymethane) in order to assess cancer susceptibility. Null mice exhibited significantly (P < 0.05) reduced levels/multiplicity of aberrant crypt foci (ACF) as compared with wild-type sibling littermate control mice. These data indicate that (i) basal/minimal dietary AA supplementation (0.6%) expands the utility of the Fads1-null mouse model for long-term cancer prevention studies and (ii) that AA content in the colonic epithelium modulates colon cancer risk. Cancer Prev Res; 9(9); 750–7. ©2016 AACR.

Wednesday, August 31, 2016 11:00 PM|Ben Néjima, Dalel; Ben Zarkouna, Yosr; Pujol, Pascal; Gammoudi, Amor; Boussen, Hamouda; Manai, Mohamed|Applied Immunohistochemistry & Molecular Morphology - Current Issue|Labels: CRC
imageTissue inhibitors of metalloproteinases (TIMPs) appear to affect many aspects of cancer biology, playing a crucial role in cell signaling by regulating cell growth, apoptosis, invasion, metastasis, angiogenesis, and genomic instability. In the present study, we investigate whether TIMP-2 SNP, TIMP-2 mRNAs, and TIMP-2 protein is associated with susceptibility to colorectal cancer (CRC) in Tunisian population. Taqman and DNA sequencing techniques were used for genotyping, TIMP-2 expression of each genotype was analyzed using semiquantitative RT-PCR and TIMP-2 protein expression was analyzed using immunohistochemistry staining. Our results showed that significantly elevated CRC risk was found in individuals with CC genotype (odds ratio 1.959; 95% confidence interval, 1.055-3.637). Moreover TIMP-2 mRNA expression in the colorectal cell carcinomas was significantly higher compared with the normal colorectal tissue (0.487±0.015 vs. 0.210±0.013) (P<0.05). In addition, serum levels of TIMP-2 were significantly lower in CRC patients than in adenoma patients (P=0.01) and healthy controls (P=0.003). Serum levels of TIMP-2 correlated significantly with tumor stage and TNM stage and were the lowest in CRC patients with stage D,T4,(N1,N2,N3),M(+). In conclusion, our study demonstrate for the first time the distribution and the clinical significance of TIMP-2 promoter polymorphisms, mRNA, protein expression, and serum level in CRC Tunisian patients suggesting that the genotyping and serum level of TIMP-2 as potential markers for susceptibility to CRC will allow a precise and early identification of individuals at high risk and will aid the design of therapeutic modalities and evaluation of treatment outcome.
Wednesday, August 31, 2016 10:05 PM|Su, L., Luo, Y., Yang, Z., Yang, J., Yao, C., Cheng, F., Shan, J., Chen, J., Li, F., Liu, L., Liu, C., Xu, Y., Jiang, L., Guo, D., Prieto, J., Avila, M. A., Shen, J., Qian, C.|Cancer Research current issue|Labels: CRC
Epithelial–mesenchymal transition (EMT) is an essential mechanism of metastasis, including in colorectal cancer. Although EMT processes are often triggered in cancer cells by their surrounding microenvironment, how EMT-relevant genes control these processes is not well understood. In multiple types of cancers, the transcription factor MEF2D has been implicated in cell proliferation, but its contributions to metastasis have not been addressed. Here, we show MEF2D is overexpressed in clinical colorectal cancer tissues where its high expression correlates with metastatic process. Functional investigations showed that MEF2D promoted cancer cell invasion and EMT and that it was essential for certain microenvironment signals to induce EMT and metastasis in vivo. Mechanistically, MEF2D directly regulated transcription of the EMT driver gene ZEB1 and facilitated histone acetylation at the ZEB1 promoter. More importantly, MEF2D responded to various tumor microenvironment signals and acted as a central integrator transducing multiple signals to activate ZEB1 transcription. Overall, our results define a critical function for MEF2D in upregulating EMT and the metastatic capacity of colorectal cancer cells. Further, they offer new insights into how microenvironment signals activate EMT-relevant genes and deepen the pathophysiologic significance of MEF2D, with potential implications for the prevention and treatment of metastatic colorectal cancer. Cancer Res; 76(17); 5054–67. ©2016 AACR.
Wednesday, August 31, 2016 6:00 PM|Song N, Pogue-Geile KL, Gavin PG, et al. |JAMA Oncology Current Issue|Labels: CRC
This secondary analysis of a randomized clinical trial investigates whether molecular subtypes of colon cancer are associated with differential prognosis and benefit from oxaliplatin added to fluorouracil plus leucovorin therapy.
Wednesday, August 31, 2016 6:00 PM|Song M, Nishihara R, Cao Y, et al. |JAMA Oncology Current Issue|Labels: CRC
This prospective cohort analysis evaluates the association between intake of marine ω-3 polyunsaturated fatty acids and risk of colorectal cancer characterized by immune infiltrate.
Friday, August 26, 2016 1:20 PM|Elsevier|JournalTOCs API - Biochemical Pharmacology (50 articles)|Labels: CRC

Rational design of an AKR1C3-resistant analog of PR-104 for enzyme-prodrug therapy

Biochemical Pharmacology, Vol. , No. (2016) pp. -
Publication date: 15 September 2016 Source:Biochemical Pharmacology, Volume 116 Author(s): Alexandra M. Mowday, Amir Ashoorzadeh, Elsie M. Williams, Janine N. Copp, Shevan Silva, Matthew R. Bull, Maria R. Abbattista, Robert F. Anderson, Jack U. Flanagan, Christopher P. Guise, David F. Ackerley, Jeff B. Smaill, Adam V. Patterson The clinical stage anti-cancer agent PR-104 has potential utility as a cytotoxic prodrug for exogenous bacterial nitroreductases expressed from replicating vector platforms. However substrate selectivity is compromised due to metabolism by the human one- and two-electron oxidoreductases cytochrome P450 oxidoreductase (POR) and aldo-keto reductase 1C3 (AKR1C3). Using rational drug design we developed a novel mono-nitro analog of PR-104A that is essentially free of this off-target activity in vitro and in vivo. Unlike PR-104A, there was no biologically relevant cytotoxicity in cells engineered to express AKR1C3 or POR, under aerobic or anoxic conditions, respectively. We screened this inert prodrug analog, SN34507, against a type I bacterial nitroreductase library and identified E. coli NfsA as an efficient bioactivator using a DNA damage response assay and recombinant enzyme kinetics. Expression of E. coli NfsA in human colorectal cancer cells led to selective cytotoxicity to SN34507 that was associated with cell cycle arrest and generated a robust ‘bystander effect’ at tissue-like cell densities when only 3% of cells were NfsA positive. Anti-tumor activity of SN35539, the phosphate pre-prodrug of SN34507, was established in ‘mixed’ tumors harboring a minority of NfsA-positive cells and demonstrated marked tumor control following heterogeneous suicide gene expression. These experiments demonstrate that off-target metabolism of PR-104 can be avoided and identify the suicide gene/prodrug partnership of E. coli NfsA/SN35539 as a promising combination for development in armed vectors. Graphical abstract

Friday, August 26, 2016 1:20 PM|Elsevier|JournalTOCs API - Biochemical and Biophysical Research Communications (50 articles)|Labels: CRC

Let-7a inhibits tumor cell growth and metastasis by directly targeting RTKN in human colon cancer

Biochemical and Biophysical Research Communications, Vol. , No. (2016) pp. -
Publication date: 16 September 2016 Source:Biochemical and Biophysical Research Communications, Volume 478, Issue 2 Author(s): Bin Li, Peng Chen, Yanxiang Chang, Jingpeng Qi, Hui Fu, Huifang Guo Colorectal cancer (CRC) is the third most common cancer worldwide, with high morbidity. MicroRNAs (miRNAs) are endogenous small RNAs that play important roles in regulating multiple biological and pathologic processes. The differential expression of miRNAs in CRC was first reported in 2003. Accumulated evidence indicates that lethal-7a (let-7a, miRNA) generally functions as a tumor suppressor in several human cancers. However, the role of let-7a in human colon cancer remains unclear. The aim of this study was to investigate the biological functions of let-7a and its potential role in colon cancer. We first discovered that let-7a level was significantly decreased in colon cancer tissues and cell lines (HT-29, HCT-116, LoVo, SW480, and SW620). To explore the effects of let-7a on colon cancer, let-7a over-expressed HCT-116 and SW620 cells were constructed. Further studies demonstrated that over-expressed let-7a could remarkably inhibit HCT-116 and SW620 cell growth and metastasis by directly down-regulating Rhotekin (RTKN). When RTKN was reintroduced into let-7a mimic transfected HCT-116 or SW620 cells, the inhibition effects of let-7a on colon cancer cell growth and metastasis were markedly reversed. In conclusion, our research shows that let-7a can inhibit tumor cell growth and metastasis by directly targeting RTKN in human colon cancer.

Thursday, August 25, 2016 6:43 AM|Cancer|Cancer via|Comments|Labels: CRC, patient perspective
CONCLUSIONSThere is poor compliance to National Comprehensive Cancer Network guidelines for adjuvant chemotherapy in patients with locally advanced rectal cancer after neoadjuvant chemoradiation and surgery. Adjuvant therapy appears to be independently associated with improved OS regardless of stage of disease, pathologic response, and patient factors. The greatest survival benefit was observed in patients who were complete responders. Age and comorbidities were found to be significantly associated with nonreceipt of adjuvant therapy. Improved rehabilitation and physical conditioning may improve the odds of patients receiving adjuvant therapy. Cancer 2016. © 2016 American Cancer Society. (Source: Cancer)
Tuesday, August 23, 2016 6:00 PM|Ko-Jiunn Liu, Tsu-Yi Chao, Jang-Yang Chang, Ann-Lii Cheng, Hui-Ju Ch’ang, Woei-Yau Kao, Yu-Chen Wu, Wei-Lan Yu, Tsai-Rong Chung and Jacqueline Whang-Peng|Most Recent Articles: Journal of Biomedical Science|Labels: CEA, CRC, clinical trial, IL
To better evaluate and improve the efficacy of dendritic cell (DC)-based cancer immunotherapy, we conducted a clinical study of patients with advanced colorectal cancer using carcinoembryonic antigen (CEA)-pul...
Tuesday, August 23, 2016 8:12 AM|Shigeyasu, K., Okugawa, Y., Toden, S., Boland, C. R., Goel, A.|Clinical Cancer Research Online First Articles|Labels: CRC

ABSTRACT Objective: Dysregulated expression of microRNAs (miRNAs) has emerged as a hallmark feature in human cancers. Exportin-5 (XPO5), a karyopherin family member, is a key protein responsible for transporting precursor miRNAs from the nucleus to the cytoplasm. While XPO5 is one of the key regulators of miRNA biogenesis, its functional role and potential clinical significance in colorectal cancer (CRC) remains unclear. Design: The expression levels of XPO5 were initially assessed in three genomic datasets, followed by determination and validation of the relationship between XPO5 expression and clinicopathological features in two independent CRC patient cohorts. A functional characterization of XPO5 in CRC was examined by targeted gene silencing in colorectal cancer cell lines and a xenograft animal model. Results: XPO5 is upregulated, both at mRNA and protein levels, in CRCs compared with normal tissues. High-XPO5 expression associated with worse clinicopathological features and poor survival in CRC patient cohorts. The siRNA knockdown of XPO5 resulted in reduced cellular proliferation, attenuated invasion, induction of G1/S cell-cycle arrest, and downregulation of key oncogenic miRNAs in CRC cells. These findings were confirmed in a xenograft animal model wherein silencing of XPO5 resulted in the attenuation of tumor growth. Conclusion: XPO5 acts like an oncogene in CRC by regulating the expression of miRNAs and may be a potential therapeutic target in CRC.

Monday, August 22, 2016 8:40 AM|Weng, W., Okugawa, Y., Toden, S., Toiyama, Y., Kusunoki, M., Goel, A.|Clinical Cancer Research Online First Articles|Labels: CRC, biomarker diagnostic

Purpose: Colorectal cancer ranks as the third most frequent cancer type, and its incidence continues to rise gradually worldwide, highlighting the need to identify previously unrecognized molecular events that propel development of this malignancy. Recent evidence suggests that dysregulated expression of FOX family of transcription factors may be critical in various genetic disorders as well as cancer; however, the functional and clinical significance of this pathway in colorectal cancer remains unclear.

Experimental Design and Results: Herein, we performed a systematic and comprehensive discovery step by evaluating the expression of FOX family members, and identified that FOXM1 and FOXQ1 are frequently overexpressed in colorectal cancer. We subsequently confirmed these findings in two large testing cohorts (n = 550) and an independent clinical validation cohort (n = 134), in which high expression of FOXM1 and FOXQ1 emerged as an independent prognostic factor in colorectal cancer patients. We corroborated these findings by performing functional assays in which knockdown of FOXM1 and FOXQ1 resulted in inhibited cell proliferation and suppressed migration and invasion in colorectal cancer cells. Furthermore, using bioinformatic approaches, we identified miR-342 as a novel regulator of both FOXM1 and FOXQ1. Overexpression or inhibition of miR-342 modulated the expression of both genes and contributed to phenotypic alterations in colorectal cancer cells, which was subsequently validated in a xenograft animal model.

Conclusions: Collectively, we have firstly identified FOXM1 and FOXQ1 as promising prognostic biomarkers in colorectal cancer patients, and provided novel evidence that therapeutic targeting of these genes or miR-342 may be a potential treatment approach in colorectal cancer patients. Clin Cancer Res; 1–11. ©2016 AACR.

Wednesday, August 17, 2016 10:10 AM|Budda, S. A., Girton, A., Henderson, J. G., Zenewicz, L. A.|Next in The JI|Labels: HIF, CRC, IL

IL-22 is expressed by activated lymphocytes and is important in modulation of tissue responses during inflammation. The cytokine induces proliferative and antiapoptotic pathways in epithelial cells allowing enhanced cell survival. This can have positive effects, such as in the maintenance of epithelial barriers in the gastrointestinal tract, but also negative effects, such as contributing to colorectal tumorigenesis. Because IL-22 can be dual-natured, we hypothesized that its biological activity should be tightly regulated to limit IL-22 expression to the sites of inflammation. One such environmental cue could be low oxygen, which often accompanies inflammation. We show that in CD4 T cells IL-22 expression is upregulated in hypoxia. The Il22 promoter contains a putative conserved hypoxic response element suggesting that the transcription factor HIF-1α may influence IL-22 expression. Differentiation in the presence of dimethyloxallyl glycine, a stabilizer of HIF-1α at normoxia, increased IL-22 expression. Using HIF-1α–deficient CD4 T cells, we show that hypoxic IL-22 upregulation is dependent on HIF-1α. These findings have implications on the regulation of Il22 gene expression and the presence of the cytokine in different inflammatory environments.

Sunday, August 14, 2016 10:05 PM|Kuo, W.-T., Lee, T.-C., Yu, L. C.-H.|Cancer Research recent issues|Labels: Src, CRC
Colorectal carcinogenesis is affected by overexpression of the lipopolysaccharide (LPS) receptors CD14 and TLR4, which antagonize each other by affecting epithelial cell proliferation and apoptosis. Eritoran is an investigational drug for sepsis treatment that resembles the lipid A moiety of LPS and therefore acts as a TLR4 inhibitor. In the present study, we explored the potential therapeutic uses and mechanisms of action of eritoran in reducing colon cancer progression. Eritoran administration via intracolonic, intragastric, or intravenous routes significantly reduced tumor burden in a chemically induced mouse model of colorectal carcinoma. Decreased proliferation and increased apoptosis were observed in mouse tumor cells after eritoran treatment. In vitro cultures of mouse primary tumor spheroids and human cancer cell lines displayed increased cell proliferation and cell-cycle progression following LPS challenge. This effect was inhibited by eritoran and by silencing CD14 or TLR4. In contrast, apoptosis induced by eritoran was eliminated by silencing CD14 or protein kinase Cζ (PKCζ) but not TLR4. Lastly, LPS and eritoran caused hyperphosphorylation of PKCζ in a CD14-dependent and TLR4-independent manner. Blocking PKCζ activation by a Src kinase inhibitor and a PKCζ-pseudosubstrate prevented eritoran-induced apoptosis. In summary, our work offers a preclinical proof of concept for the exploration of eritoran as a clinical treatment, with a mechanistic rationale to reposition this drug to improve the management of colorectal cancer. Cancer Res; 76(16); 4684–95. ©2016 AACR.
Sunday, August 14, 2016 10:05 PM|Becht, E., de Reynies, A., Giraldo, N. A., Pilati, C., Buttard, B., Lacroix, L., Selves, J., Sautes-Fridman, C., Laurent-Puig, P., Fridman, W. H.|Clinical Cancer Research recent issues|Labels: breast cancer, CRC

Purpose: The tumor microenvironment is formed by many distinct and interacting cell populations, and its composition may predict patients' prognosis and response to therapies. Colorectal cancer is a heterogeneous disease in which immune classifications and four consensus molecular subgroups (CMS) have been described. Our aim was to integrate the composition of the tumor microenvironment with the consensus molecular classification of colorectal cancer.

Experimental Design: We retrospectively analyzed the composition and the functional orientation of the immune, fibroblastic, and angiogenic microenvironment of 1,388 colorectal cancer tumors from three independent cohorts using transcriptomics. We validated our findings using immunohistochemistry.

Results: We report that colorectal cancer molecular subgroups and microenvironmental signatures are highly correlated. Out of the four molecular subgroups, two highly express immune-specific genes. The good-prognosis microsatellite instable–enriched subgroup (CMS1) is characterized by overexpression of genes specific to cytotoxic lymphocytes. In contrast, the poor-prognosis mesenchymal subgroup (CMS4) expresses markers of lymphocytes and of cells of monocytic origin. The mesenchymal subgroup also displays an angiogenic, inflammatory, and immunosuppressive signature, a coordinated pattern that we also found in breast (n = 254), ovarian (n = 97), lung (n = 80), and kidney (n = 143) cancers. Pathologic examination revealed that the mesenchymal subtype is characterized by a high density of fibroblasts that likely produce the chemokines and cytokines that favor tumor-associated inflammation and support angiogenesis, resulting in a poor prognosis. In contrast, the canonical (CMS2) and metabolic (CMS3) subtypes with intermediate prognosis exhibit low immune and inflammatory signatures.

Conclusions: The distinct immune orientations of the colorectal cancer molecular subtypes pave the way for tailored immunotherapies. Clin Cancer Res; 22(16); 4057–66. ©2016 AACR.

Sunday, August 14, 2016 10:05 PM|Dunne, P. D., McArt, D. G., Bradley, C. A., O'Reilly, P. G., Barrett, H. L., Cummins, R., O'Grady, T., Arthur, K., Loughrey, M. B., Allen, W. L., McDade, S. S., Waugh, D. J., Hamilton, P. W., Longley, D. B., Kay, E. W., Johnston, P. G., Lawler, M., Salto-Tellez, M., Van Schaeybroeck, S.|Clinical Cancer Research recent issues|Labels: CRC

Purpose: A number of independent gene expression profiling studies have identified transcriptional subtypes in colorectal cancer with potential diagnostic utility, culminating in publication of a colorectal cancer Consensus Molecular Subtype classification. The worst prognostic subtype has been defined by genes associated with stem-like biology. Recently, it has been shown that the majority of genes associated with this poor prognostic group are stromal derived. We investigated the potential for tumor misclassification into multiple diagnostic subgroups based on tumoral region sampled.

Experimental Design: We performed multiregion tissue RNA extraction/transcriptomic analysis using colorectal-specific arrays on invasive front, central tumor, and lymph node regions selected from tissue samples from 25 colorectal cancer patients.

Results: We identified a consensus 30-gene list, which represents the intratumoral heterogeneity within a cohort of primary colorectal cancer tumors. Using a series of online datasets, we showed that this gene list displays prognostic potential HR = 2.914 (confidence interval 0.9286–9.162) in stage II/III colorectal cancer patients, but in addition, we demonstrated that these genes are stromal derived, challenging the assumption that poor prognosis tumors with stem-like biology have undergone a widespread epithelial–mesenchymal transition. Most importantly, we showed that patients can be simultaneously classified into multiple diagnostically relevant subgroups based purely on the tumoral region analyzed.

Conclusions: Gene expression profiles derived from the nonmalignant stromal region can influence assignment of colorectal cancer transcriptional subtypes, questioning the current molecular classification dogma and highlighting the need to consider pathology sampling region and degree of stromal infiltration when employing transcription-based classifiers to underpin clinical decision making in colorectal cancer. Clin Cancer Res; 22(16); 4095–104. ©2016 AACR.

See related commentary by Morris and Kopetz, p. 3989

Sunday, August 14, 2016 10:05 PM|Wei, P.-L., Tseng, C.-S., Chen, C.-C., Chiu, C.-J., Hsiao, G.|Clinical Cancer Research recent issues|Labels: CRC

Background: Previously we have demonstrated the novel microtube array membrane (MTAM) can serves as an excellent substrate for a rapid, in vivo anti-cancer drug screening assay based on the hollow fiber assay (HFA), with higher cell-drug sensitivity, cell-host interaction, plus the clear angiogenesis, owing to its unique structural characteristics. In current study, the patient derived tumor cells (PDTC) were subjected to MTAM/X based screening platform. Patient derived colon cancer sample was treated and loaded into MTAM, studied first in cell growth and characterization, and later the cell toxicity by several representative cancer drugs, both in vitro and in vivo. The goal of this study is to explore the potential of utilizing MTAM/X with PDX for rapid, reliable, lower cost personalized anti-cancer drug selection platform.

Materials and Methods: Nano-porous PLLA MTAMs (npMTAM) were fabricated via a co-axial electrospinning and characterized morphologically first, then mechanical and permeation properties were determined. The nano-porous microstructure was also characterized by porosimeter. PTDC (colon cancer, 0.5 cm3) was obtained surgically and treated with enzyme-containing medium, homogenized before loaded into npMTAM at 105-106 cell per sample. The subcutaneous implantation of these MTAMs were conducted with 4 MTAMs per model (BALB/c) one day before the drug treatment. Regiments used according to the recommended dosage were cisplatin, 5 FU and Erbitux. In vitro and in vivo PDTC growth within MTAM were determined via MTT assay. The in vivo drug cytotoxicity towards PDTC was then determined by MTT in a 9-day period, in the meantime, the supportive results of angiogenesis was characterized via imaging process.

Results: Nano-porous PLLA MTAMs were successfully prepared with a tube wall thickness of 2-3 microns and pore size of 100-200 nm. PDTC (colon cancer) was loaded into npMTAM within 2 hours after surgical collection. The in vitro and in vivo growth of PDTC in npMTAM was observed clearly in a two weeks period. Upon the administration of 5 FU, Cisplatin and Erbitux to the test model, cell growth was altered, however in different fashion. 5FU show early cell cytotoxicity, however, loss its inhibition capability afterwards. Erbitux showed no sign of inhibition. Finally greatest inhibition of this particular patient tumor was found with cisplatin. Extent of angiogenesis was found to be in consist with MTT data. The overall time needed to generate the final results is within 2 weeks.

Conclusions: Testing PDTC in vivo within npMTAM based assay (PDXiMTAM), we demonstrated the feasibility of in vivo personalized cancer drug selection platform can be achieved in a reasonable time periods of 2 weeks. Selection from a group of four regiments, cisplatin demonstrated its highest cytotoxicity towards this specific tumor. This will be a very practical assay in clinical setting to identify most effective drug at personal level. Our results therefore suggest that such assay could be further developed in timely treatment selection. A double blind, cohort clinical trial of 60 patients with colon cancer is currently ongoing to validate PDXiMTAM.

Citation Format: Po-Li Wei, Chia-Shiuan Tseng, Chien-Chung Chen, Chien-Jin Chiu, George Hsiao. The implication of patient derived tumor cell (PDTC) tested/screened with novel microtube array membrane (MTAM)-based hollow fiber assay (HFA).. [abstract]. In: Proceedings of the AACR Special Conference: Patient-Derived Cancer Models: Present and Future Applications from Basic Science to the Clinic; Feb 11-14, 2016; New Orleans, LA. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(16_Suppl):Abstract nr A33.

Sunday, August 14, 2016 10:05 PM|Sloma, I., Ben-zvi, I., Khor, T., Ciznadija, D., Katz, A., Vasquez, D., Jaskowiak, J., Ryland, L., Davies, A., Sidransky, D., Paz, K.|Clinical Cancer Research recent issues|Labels: CRC

Background: Patient-derived xenograft (PDX) models, also known as Champions TumorGraft® models, maintain the complex intra-tumoral biology of the primary tumor. Over 250 of the Champions models, ranging over a wide variety of solid tumors and passaging generations, have been analyzed using whole exome sequencing (WES) and RNA sequencing (RNAseq). SNPs, InDels and copy number alterations (CNAs) data have been generated for each model, following the Genome Analysis Toolkit (GATK). While several publications compare small numbers of PDX models and human tumors on the molecular level, this is the first known comprehensive analysis whereby the molecular fidelity of the PDX platform is corroborated across several cancer types and throughout different mouse generations.

Method and Results: First, we compared PDXs to their human original counterparts using a preliminary group of four PDX models with available matching human patient WES data. Patient tumor source included dedifferentiated liposarcoma, synovial sarcoma, renal cell carcinoma and squamous cell carcinoma of the lung. PDX passages ranged from 2 to 4. We compared called mutations and a high percentage of identified human tumor mutations were present in the PDX models (42-82%), with the lowest scoring model also showing signs of normal contamination in the human tumor sample. For CNAs in oncogenic sites, we saw an average of 65% of human tumor alterations recurring in the PDX models. This was observed, despite inherent difficulties due to exome- based CNA analysis methods.

Encouraged by the individual patient results, we subjected our largest (per cancer type) PDX cohorts to a molecular comparison with the equivalent TCGA cohorts. More than 200 of the sequenced models, grouped into colorectal adenocarcinoma (COADREAD), lung adenocarcinoma (LUAD), breast carcinoma (BRCA), head and neck squamous cell carcinoma (HNSC) and ovarian serous carcinoma (OV) cohorts were compared. We applied mutation category (MC) and significantly mutated genes (SMG) analysis, as well as comparison of mutation population frequencies for TCGA SMG. Results showed high correlation between the TCGA and the Champions PDX cohorts, although the level of matching varied between cancer types. For instance, COADREAD was highly correlative, while other cancer types, such as BRCA, showed bias toward CpG site mutations. In SMG analysis and population frequency analysis, major SMGs recur across the cohorts, while, as expected, weaker signals from the TCGA were often missed in the smaller cohorts.

Conclusions: Detailed comparison of several PDX models to the human tumor counterpart demonstrated high fidelity, not only at the gene level but also the mutation and CNA level. Cohort comparisons were correlative as well, but a certain bias was discerned in both MC and SMG analyses. There could be several causes for this, including statistical artifacts due to small cohort sizes, clinical and demographic differences between the Champions and TCGA patient profiles, or biological factors such as clonal selection and engraftment pressure. Further analysis is ongoing to better understand the model at a molecular level and maximize its utility as a robust translational research tool.

Citation Format: Ido Sloma, Ido Ben-zvi, Tin Khor, Daniel Ciznadija, Amanda Katz, David Vasquez, Jennifer Jaskowiak, Lindsay Ryland, Angela Davies, David Sidransky, Keren Paz. Accurate molecular fidelity of patient-derived xenograft (PDX) models to original human tumors and to The Cancer Genome Atlas (TCGA). [abstract]. In: Proceedings of the AACR Special Conference: Patient-Derived Cancer Models: Present and Future Applications from Basic Science to the Clinic; Feb 11-14, 2016; New Orleans, LA. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(16_Suppl):Abstract nr A21.

Sunday, August 14, 2016 10:05 PM|Dangles-Marie, V.|Clinical Cancer Research recent issues|Labels: CRC

Through 2 examples of digestive tumors, (colorectal cancer, "CRC", and anal squamous cell carcinoma, "ASCC"), we will present the development and use of patient-derived tumor xenografts.

Colorectal cancer remains a major cause of mortality worldwide and CRC patient death is generally attributable to metastasis development. By contrast, ASCC is a rare tumor (< 5% of all lower gastrointestinal tract malignancies in Europe) but with increasing incidence and mutilating surgery for patients with chemoradiotherapy-resistant tumors. Consequently, these both digestive tumors require new therapeutic approaches and relevant preclinical models to identify and test new drugs/combinations. In this context, we develop and characterize patient-derived CRC and ASCC xenografts.

We will illustrate the capacity of these PDXs to recapitulate the intratumoral clonal heterogeneity, gene expression profile, key molecular alterations, histology and drug response of the patient tumors through passages in mice. In addition to these now well-known features depicted in a large number of tumor types, we will discuss about practical situations (orthotopic engraftment, impact of mouse host, criteria for measuring drug efficacy). Finally, we will report alternative methods as ex vivo tumor spheres we have developed.

Citation Format: Virginie Dangles-Marie. PDX in colorectal cancer and carcinoma of the anal canal. [abstract]. In: Proceedings of the AACR Special Conference: Patient-Derived Cancer Models: Present and Future Applications from Basic Science to the Clinic; Feb 11-14, 2016; New Orleans, LA. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(16_Suppl):Abstract nr IA05.

Sunday, August 14, 2016 10:05 PM|Maekawa, H., Yamaura, T., Kawada, K., Sakai, Y.|Clinical Cancer Research recent issues|Labels: CRC

Colorectal cancer (CRC) is one of the most common cancers worldwide, with more than 690,000 deaths estimated in 2012. Despite our increasing knowledge of the genetics of CRC, few biomarkers are available for successful anticancer therapies. The key drugs and therapeutic strategy against CRC are limited. The overall 5-year survival rate of patients with metastatic CRC is only ~12%.

Because of the heterogeneity of CRCs and the complexity in clinical conditions, traditional cell line models have not predicted clinical outcomes of anticancer therapies. Recent studies show that it is difficult to associate the specific genetic mutations with drug sensitivities in vitro. On the other hand, patient-derived tumor xenografts (PDXs) are preferable preclinical models as they preserve the genomic condition and tumor heterogeneity in primary lesions. Although the PDX is costly and time-consuming, its usefulness outweighs these limitations, and it is expected to help not only basic cancer studies but also development of personalized therapies in the clinical setting.

We have been establishing PDXs of advanced CRCs. In short, we dissect fresh tumor fragments (3-5mm cube) from resected tumors by surgical procedure after obtaining patients' informed consents in our hospital. We bring them to our laboratory on ice. We then wash tumor samples with medium twice and with PBS twice. The tumor samples are implanted to each flank of nude mice and/or NOD/SCID mice. The implanted tumor growth is measured at least once a week. We consider the implantation is successful when estimate tumor volumes become more than 1000 mm3. We excise tumor masses, cut and divide them into small fragments, and perform serial transplantation.

We have engrafted 50 cases of CRC tumors so far to establish PDXs. In 23 cases, we confirmed that engrafted tumors were successfully growing. In 8 cases, the implanted tumor disappeared within several weeks after transplantation, and we determined that PDXs in these cases failed to grow as any tumor masses were developed for 6 months. The remaining 19 cases are under study. We found that each PDX, even from the same tumor, showed different growth rate, which made it difficult to set-up PDX panels for drug-sensitivity tests. We conclude that it is rather difficult to prepare an enough number of homogeneous PDX models from each cancer for evaluation of its chemosensitivity. We are currently trying to minimize the growth rate differences of PDXs among individual transplants from a same patient. Such a technique may accelerate translational study using PDXs.

Our future aim is to optimize the care for individual patients with advanced CRC by selecting effective regimens that are predicted by the chemosensitivity test using PDXs. We will also study several genetic mutations that affect patient survival rate and their relationship with chemo-sensitivities so that we can improve CRC patient survival in our hospital.

Citation Format: Hisatsugu Maekawa, Tadanori Yamaura, Kenji Kawada, Yoshiharu Sakai. Summary of 50 cases of patient-derived colorectal cancer xenografts; Problems and tips to obtain appropriate results in translational researches. [abstract]. In: Proceedings of the AACR Special Conference: Patient-Derived Cancer Models: Present and Future Applications from Basic Science to the Clinic; Feb 11-14, 2016; New Orleans, LA. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(16_Suppl):Abstract nr B36.

Sunday, August 14, 2016 10:05 PM|Medico, E.|Clinical Cancer Research recent issues|Labels: CRC

Tumor-stroma interactions have long been implicated in neoplastic aggressiveness and resistance to treatment. In PDX models, after engraftment, the stromal components are contributed by the mouse host. As a consequence, transcripts expressed by cancer cells can be distinguished from stromal transcripts by their species of origin. We therefore exploited species-specific expression profiling of colorectal cancer (CRC) PDXs to evaluate the involvement of stromal components in recently defined CRC transcriptional subtypes. We found that stroma-derived transcripts profoundly affect the overall tumor transcriptome, and that a previously defined poor prognosis subtype is actually endowed with abundant stroma. To stratify CRC based on cancer cell-intrinsic transcriptional features, we performed class discovery on a human-specific gene expression dataset of 515 CRC PDXs, and identified five CRC intrinsic subtypes (CRIS). CRIS classification successfully categorized independent sets of primary and metastatic CRC tumors, with limited overlap on existing transcriptional classes and unprecedented predictive and prognostic performances. In a complementary approach, we focused on stroma-specific transcripts to define expression signatures reflecting the abundance in human CRC samples of fibroblasts, leukocytes and endothelial cells. Stromal scores were found to correlate with poor CRC prognosis and resistance of rectal cancer to preoperative radiotherapy, highlighting the microenvironment as a variable and dynamic component of CRC, with diagnostic and therapeutic potential.

Citation Format: Enzo Medico. Expression profiling of colorectal cancer PDX models for disease stratification and tumor-stroma interaction analysis. [abstract]. In: Proceedings of the AACR Special Conference: Patient-Derived Cancer Models: Present and Future Applications from Basic Science to the Clinic; Feb 11-14, 2016; New Orleans, LA. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(16_Suppl):Abstract nr IA09.

Sunday, August 14, 2016 10:05 PM|McDermott, J. E., Liu, T., Petyuk, V., Smith, R., Rodland, K.|Clinical Cancer Research recent issues|Labels: CRC

Patient-derived xenografts (PDXs) are an important and powerful tool to study many types of cancer and provide valuable insight into potential drug response and development of resistance. Due to the differences in tissue lineage PDXs also allow the dissection of stromal contribution to the tumor that is not easily obtainable from examination of the original tumor itself. We have applied mass-spectrometry enabled proteomics, phosphoproteomics, and RNAseq to compare primary metastasis, and PDXs derived from several metastatic sites from the same original tumor which was a colorectal adenocarcinoma.

Comparing our results with a large-scale proteomic characterization of tumors performed under the clinical tumor analysis consortium (CPTAC) shows that the original metastasis, and PDXs from different sites, are significantly similar, regardless of metastasis site. Proteomic profiles of these samples are enriched in proteins expressed in the tissue of origin, rather than the site of metastasis. Pathway analysis from proteomics revealed enrichment in complement cascade, extracellular matrix receptors, and focal adhesion. These were different than the pathways indicated by the transcriptomics, which included cell cycle, focal adhesion, and MAP signaling, but not complement or extracellular matrix.

We found in the case of complement and focal adhesion the original metastasis sample was very different from the PDX samples, if we only considered human protein contributions. Considering mouse proteins we found that the mouse stromal contribution complemented the human tumor contribution in the PDXs, aligning the functional analysis in both types of samples. Combining these results with subsequent analysis of driver mutations revealed that the proteomic contribution of stroma was as high as 40%. We have extended these observations to additional pairs of clinical tumor specimens and paired PDXs, including different tumor types. In all cases, there is greater similarity among the same sample type (resected tumor, metastatic lesion, or PDX) than between different sample types from the same patient.

Citation Format: Jason E. McDermott, Tao Liu, Vladislav Petyuk, Richard Smith, Karin Rodland. Proteomic characterization of stromal contribution to tumor using patient-derived xenografts. [abstract]. In: Proceedings of the AACR Special Conference: Patient-Derived Cancer Models: Present and Future Applications from Basic Science to the Clinic; Feb 11-14, 2016; New Orleans, LA. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(16_Suppl):Abstract nr A31.

Friday, August 12, 2016 12:58 PM|Bartolini, A., Cardaci, S., Lamba, S., Oddo, D., Marchio, C., Cassoni, P., Amoreo, C. A., Corti, G., Testori, A., Bussolino, F., Pasqualini, R., Arap, W., Cora, D., Di Nicolantonio, F., Marchio, S.|Clinical Cancer Research Online First Articles|Labels: RAS, CRC

Purpose: KRAS mutations confer adverse prognosis to colorectal cancer, and no targeted therapies have shown efficacy in this patient subset. Paracrine, nongenetic events induced by KRAS-mutant tumor cells are expected to result in specific deregulation and/or relocation of tumor microenvironment (TME) proteins, which in principle can be exploited as alternative therapeutic targets.

Experimental Design: A multimodal strategy combining ex vivo/in vitro phage display screens with deep-sequencing and bioinformatics was applied to uncover TME-specific targets in KRAS-mutant hepatic metastasis from colorectal cancer. Expression and localization of BCAM and LAMA5 were validated by immunohistochemistry in preclinical models of human hepatic metastasis and in a panel of human specimens (n = 71). The antimetastatic efficacy of two BCAM-mimic peptides was evaluated in mouse models. The role of BCAM in the interaction of KRAS-mutant colorectal cancer cells with TME cells was investigated by adhesion assays.

Results: BCAM and LAMA5 were identified as molecular targets within both tumor cells and TME of KRAS-mutant hepatic metastasis from colorectal cancer, where they were specifically overexpressed. Two BCAM-mimic peptides inhibited KRAS-mutant hepatic metastasis in preclinical models. Genetic suppression and biochemical inhibition of either BCAM or LAMA5 impaired adhesion of KRAS-mutant colorectal cancer cells specifically to endothelial cells, whereas adhesion to pericytes and hepatocytes was unaffected.

Conclusions: These data show that the BCAM/LAMA5 system plays a functional role in the metastatic spreading of KRAS-mutant colorectal cancer by mediating tumor–TME interactions and as such represents a valuable therapeutic candidate for this large, currently untreatable patient group. Clin Cancer Res; 1–11. ©2016 AACR.

Tuesday, August 9, 2016 1:03 PM|Toffoli, G., Sharma, M. R., Marangon, E., Posocco, B., Gray, E., Mai, Q., Buonadonna, A., Polite, B. N., Miolo, G., Tabaro, G., Innocenti, F.|Clinical Cancer Research Online First Articles|Labels: CRC, clinical trial

Purpose:UGT1A1*28 confers a higher risk of toxicity in patients treated with irinotecan. Patients with *1/*1 and *1/*28 genotypes might tolerate higher than standard doses of irinotecan. We aimed to identify the maximum tolerated dose (MTD) of irinotecan in mCRC patients with *1/*1 and *1/*28 genotypes treated with FOLFIRI plus bevacizumab, and to determine whether bevacizumab alters irinotecan pharmacokinetics. Experimental Design: Previously untreated mCRC patients (25 *1/*1; 23 *1/*28) were given FOLFIRI plus bevacizumab every two weeks. The irinotecan dose was escalated using a 3+3 design in each genotype group as follows: 260, 310, 370 mg/m2. The MTD was the highest dose at which < 4/10 patients had a dose-limiting toxicity (DLT). Pharmacokinetics of irinotecan and SN-38 were measured on days 1-3 (without bevacizumab) and 15-17 (with bevacizumab). Results:For *1/*1 patients, 2 DLTs were observed among 10 patients at 310 mg/m2, while 370 mg/m2 was not tolerated (2 DLTs in 4 patients). For *1/*28 patients, 2 DLTs were observed among 10 patients at 260 mg/m2, while 310 mg/m2 was not tolerated (4 DLTs in 10 patients). Neutropenia and diarrhea were the most common DLTs. Changes in the AUCs of irinotecan and SN-38 associated with bevacizumab treatment were marginal. Conclusions:The MTD of irinotecan in FOLFIRI plus bevacizumab is 310 mg/m2 for UGT1A1 *1/*1 patients and 260 mg/m2 for *1/*28 patients. Bevacizumab does not alter the pharmacokinetics of irinotecan. The antitumor efficacy of these genotype-guided doses should be tested in future studies of mCRC patients treated with FOLFIRI plus bevacizumab.

Thursday, August 4, 2016 6:00 PM|Chenli Li, Lingxu Zhao, Yuan Chen, Tiantian He, Xiaowan Chen, Jiating Mao, Chunmei Li, Jianxin Lyu and Qing H. Meng|Articles: MD Anderson Cancer Center|Labels: CRC
MicroRNA-21 (miR-21) is up-regulated in many cancers, including colorectal cancer (CRC). Nevertheless, the function of miR-21 in CRC and the mechanism underlying that function is still unclear.
Tuesday, August 2, 2016 6:00 PM|Ken Cadigan|Cancers|Labels: WNT, CRC
T-cell Factor/Lymphoid Enhancer Factor (TCF/LEF) transcription factors are major regulators of Wnt targets, and the products of the TCF7 and TCF7L2 genes have both been implicated in the progression of colorectal cancer in animal models and humans. TCFs recognize specific DNA sequences through their high mobility group (HMG) domains, but invertebrate TCFs and some isoforms of vertebrate TCF7 and TCF7L2 contain a second DNA binding domain known as the C-clamp. This review will cover the basic properties of C-clamps and their importance in Wnt signaling, using data from Drosophila, C. elegans, and mammalian cell culture. The connection between C-clamp containing TCFs and colorectal cancer will also be discussed.
Monday, August 1, 2016 3:50 AM|Goro Nakayama, Tsutomu Fujii, Kenta Murotani, Keisuke Uehara, Norifumi Hattori, Masamichi Hayashi, Chie Tanaka, Daisuke Kobayashi, Mitsuro Kanda, Suguru Yamada, Hiroyuki Sugimoto, Masahiko Koike, Michitaka Fujiwara, Yuichi Ando, Yasuhiro Kodera|Cancer Science|Labels: CRC
The identification of surrogate markers for long-term outcomes in patients with metastatic colorectal cancer (mCRC) may help in designing treatment regimens. The aim of this study was to assess whether two-dimensional response (2-DR) can serve as a new surrogate marker for overall survival (OS) in patients with mCRC. The study population consisted of 99 patients with mCRC from two independent cohorts who were treated with oxaliplatin-based chemotherapy plus bevacizumab. 2-DR was defined as an area enclosed by coordinate points, including early tumour shrinkage (ETS) at 8 weeks, depth of response (DpR) at nadir and 20% increase over nadir at progression. Each variable was weighted by its contribution rate to OS. The model was developed and internally validated in the learning cohort, and the performance of this model was externally verified in the validation cohort. Spearman correlation coefficients for 2-DR and OS in the learning and validation cohorts were 0.593 and 0.661, respectively. The C-indexes in predicting OS were 0.724 (95% CI: 0.623–0.815) in the learning cohort and 0.762 (95% CI: 0.651–0.873) in the validation cohort. OS was significantly longer in patients with high 2-DR values than in patients with low 2-DR values in both the learning (37.0 versus 24.1 months, p<0.001) and validation (41.2 versus 20.4 months, p<0.001) cohorts. In contrast, differences in ETS and DpR were not statistically significant. Multivariate analyses showed that 2-DR was an independent prognostic factor for OS. This article is protected by copyright. All rights reserved.
Thursday, July 28, 2016 1:46 PM|Onclive Colorectal Cancer Articles|Labels: FGFR, PDGF, VEGF, CRC
Axel Grothey, MD, discusses both the LUME-1 and LUME-2 trials, the differences between left and right tumors in colorectal cancer, and how that information could potentially be used in diagnosis and treatment.
Thursday, July 28, 2016 7:56 AM|Flinders, C., Liu, S., Mumenthaler, S.|Cancer Research recent issues|Labels: CRC
Introduction: Metastasis is responsible for over 90% of cancer related mortality; however, somatic mutations unique to the metastatic phenotype have yet to be identified. This raises the possibility that the metastatic phenotype is the result of a transcriptional profile adopted by cells of the primary tumor enabling them to spread to distal regions of the body. The tumor microenvironment (TME) is capable of influencing gene expression and cellular behavior. Hypoxia is a component of the TME that arises due to improper vascular architecture, which leads to reduced oxygen delivery to tissues and correlates with poor survival outcome. Stabilization of the transcription factor HIF-1α during hypoxia induces the expression of genes involved in metastasis through direct activation or through the alteration of the epigenome. However, upon re-oxygenation, HIF-1α is degraded and can no longer induce expression of hypoxic responsive genes. During metastasis, cells that have adopted hypoxia induced metastatic behavior such as increased motility and invasiveness must transit through regions of normoxia on their way to metastatic colonization.Hypothesis: As hypoxic cells migrate away from regions of low oxygen toward oxygen rich environments (e.g. venous blood) genes induced by hypoxia would be expected to return to basal levels of expression. Those genes whose expression does not return to levels associated with normoxia, but instead remain elevated following re-oxygenation may be crucial for maintaining the hypoxia induced metastatic phenotype.Experimental Design: To study the stability of hypoxia induced gene expression alterations following re-oxygenation, colon cancer cell lines were grown under hypoxic conditions (1% O2) for up to 3 weeks and then transitioned back to normoxic conditions. Gene expression changes were assessed during hypoxia adaptation and for 3 additional weeks following re-oxygenation.Results: The continued expression of hypoxia induced genes following re-oxygenation varied greatly among those tested. Gene transcription associated with epithelial-to-mesenchymal transition (EMT) such as VIM and ZEB1 were induced by 3 weeks of hypoxic conditioning and remained elevated following their return to normoxic culturing conditions. However, in contrast, hypoxia induced genes involved in the Warburg effect such as PDK1 were induced during hypoxia, yet quickly returned to at or below basal levels following re-oxygenation.Conclusion: Continued expression of hypoxia induced genes following re-oxygenation varies from gene to gene and from pathway to pathway. There appears to be some consistency in expression of genes following re-oxygenation that are involved in migration and invasion such as ZEB1. These results suggest that certain pathways such as EMT may be vital for the maintenance of the hypoxia induced metastatic phenotype as these cells transit from regions of low oxygen to regions of higher or normal oxygen levels.Citation Format: Colin Flinders, Sonya Liu, Shannon Mumenthaler. Stability of hypoxia induced expression of colon cancer metastasis-associated genes. [abstract]. In: Proceedings of the AACR Special Conference: Function of Tumor Microenvironment in Cancer Progression; 2016 Jan 7–10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2016;76(15 Suppl):Abstract nr C08.
Thursday, July 28, 2016 7:56 AM|Kehlet, S. N., Sanz-Pamplona, R., Leeming, D. J., Karsdal, M. A., Moreno, V.|Cancer Research recent issues|Labels: CRC, biomarker diagnostic
Background: The local microenvironment of a tumor plays an important role in colorectal cancer development. One of the hallmarks of colorectal cancer is abnormal remodeling of the extracellular matrix within the tumor microenvironment resulting in increased collagen deposition (desmoplasia) and increased matrix metalloproteinase (MMP) activity. This leads to the release of protein degradation fragments into the circulation.We investigated the potential of disease relevant fragments of collagen type I, III and IV as novel serological biomarkers for colorectal cancer.Methods: The level of biomarkers reflecting MMP-mediated degradation of type I (C1M), type III (C3M) and type IV (C4M) collagen and the formation of type III (Pro-C3) collagen were measured in serum from patients diagnosed with colorectal cancer (n=196, stage I-IV), subjects with adenomas (n=99, low risk-high risk) and age matched healthy controls (n=99). The levels of the individual biomarkers were compared using ANCOVA to adjust for age and gender on normalized data. Area under the receiver operating characteristics (AUROC) and logistic regression modeling were carried out to evaluate the discriminative power of the biomarkers.Results: Circulating levels of C1M, C3M and Pro-C3 were significantly elevated in colorectal cancer patients compared to subjects with adenomas (C1M and Pro-C3: p<0.0001, C3M: p<0.01) and healthy controls (C1M and Pro-C3: p<0.01, C3M: p<0.05). When patients were stratified according to their stage, all four biomarkers were able to differentiate stage IV patients from all other stages, adenomas and healthy controls (p<0.05-0.0001). Pro-C3 and C1M had the highest diagnostic accuracy in relation to identifying metastatic tumors from non-metastatic tumors with an AUROC of 0.80 and 0.78, respectively. When all biomarkers were combined the diagnostic power was improved (AUROC=0.85).Conclusion: In the present study we showed that specific serum biomarkers, reflecting collagen remodeling in the tumor microenvironment, could separate metastatic colorectal cancer patients from non-metastatic patients with high diagnostic power. This suggests that ECM remodeling is a key event in colorectal cancer development and biomarkers reflecting altered disease pathogenesis may increase the understanding of disease mode of action and provide a clinical tool for stratifying and monitoring patients according to disease severity.Citation Format: Stephanie Nina Kehlet, Rebeca Sanz-Pamplona, Diana Julie Leeming, Morten Asser Karsdal, Victor Moreno. Serological biomarkers reflecting collagen remodeling of the tumor microenvironment are elevated in metastatic colorectal cancer patients. [abstract]. In: Proceedings of the AACR Special Conference: Function of Tumor Microenvironment in Cancer Progression; 2016 Jan 7–10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2016;76(15 Suppl):Abstract nr C16.
Thursday, July 28, 2016 7:56 AM|Rasanen, K., Lehtinen, E., Nokelainen, K., Hautala, L., Itkonen, O., Stenman, U., Koistinen, H.|Cancer Research recent issues|Labels: CRC, clinical trial, IL
Inflammation is known to promote colorectal cancer (CRC) tumorigenesis, but the underlying molecular mechanisms are still being uncovered. Proinflammatory cytokine interleukin-6 (IL-6) is primarily secreted by the cells of the tumor microenvironment (TME), mainly by inflammatory cells but also by activated fibroblasts. IL-6 stimulates growth and survival signaling in CRC. Inflammatory signals regulate also the production and activity of proteases and their inhibitors. Serine protease inhibitor Kazal type1 (SPINK1, aka tumor-associated trypsin inhibitor, TATI) is expressed in several tissues and over-expression of SPINK1 predicts an unfavorable outcome in many cancers, including colon cancer. The SPINK1 gene contains an IL-6 responsive element and in addition to being a protease inhibitor, it also acts as an acute phase reactant and a growth factor. Expression of serine proteases trypsin-1 and -2, the main targets of SPINK1, also correlate with malignancy and metastatic potential. The aim of this study was to assess the paracrine signaling between fibroblast-derived IL-6 and cancer cell-derived SPINK1, trypsin-1 and -2. The following expression analyses were used: quantitative PCR, immunohistochemistry, immunofluorometric assays and Western blotting. The results show that CRC cells Colo205 and HT-29 express SPINK1 and secrete it into the culture medium. IL-6 dose-dependently increased the mRNA expression and protein levels of SPINK1. Conditioned media from fibroblasts had the same effect, and conversely CRC media increased IL-6 secretion in the fibroblasts, indicating paracrine signaling between these cell types. In CRC tissues cancer cells were positive for SPINK1, while IL-6 was found in fibroblasts surrounding the cancer cells, confirming the in vitro results. In Colo205 cells the baseline levels of trypsin-1 and -2 and the respective genes PRSS1 and PRSS2 were much higher compared to HT-29 cells. In Colo205 cells IL-6 led to concomitant increase in the secretion of trypsin-1 and -2, whereas in HT-29 cells these remained constantly low. Mechanistically, addition of IL-6 led to activation of the canonical Stat3 pathway, as indicated by Stat3 phosphorylation. Stat3 inhibitor reduced both SPINK1 and trypsin-1 and -2 levels, demonstrating that Stat3 is the transcription factor driving their expression. Taken together, our results show a connection between TME-derived inflammatory response and increased SPINK1 and trypsin-1 and -2 levels. Elevated serum SPINK1 has been shown to be an independent prognostic factor in colon cancer. As SPINK1 acts as a growth factor in some cancers, it has also been suggested as a therapeutic target. Therefore assessment of SPINK1 expression and function has several potentially important clinical applications in colorectal and other cancers. The study also strengthens the role of tumor microenvironment in tumor progression and emphasizes the importance of studying tumor-stroma crosstalk of proteolytic processing.Citation Format: Kati Räsänen, Elina Lehtinen, Kristiina Nokelainen, Laura Hautala, Outi Itkonen, Ulf-Håkan Stenman, Hannu Koistinen. Interleukin-6 induces secretion of SPINK1 and trypsin in colorectal cancer. [abstract]. In: Proceedings of the AACR Special Conference: Function of Tumor Microenvironment in Cancer Progression; 2016 Jan 7–10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2016;76(15 Suppl):Abstract nr A42.
Thursday, July 28, 2016 7:56 AM|Preet, R., Zhuang, S., Hung, W.-T., Christenson, L. K., Dixon, D. A.|Cancer Research recent issues|Labels: CRC
Enhanced secretion of extracellular vesicles such as microvesicles and exosomes by cancer cells has been recently recognized as a new means of transferring oncogenic information within the tumor microenvironment. Through their ability to carry specific RNA and protein cargo, tumor-derived exosomes can directly impact neighboring cells to support tumor progression. However, it remains unclear what regulates exosome production in tumor cells and the factors influencing loading of tumor-promoting RNA cargo. Our prior work has established that colorectal cells (CRC) and tumors overexpress the key RNA-binding protein HuR early in GI tumor development. When overexpressed and present in the cytoplasm, HuR can promote mRNA stabilization of tumor-promoting genes through binding of 3'UTR AU-rich elements (ARE). While the ability of HuR overexpression to promote gene expression is well recognized, our understanding of how HuR communicates this information within the tumor microenvironment is limited. To test if HuR overexpression could impact exosomes, Tet-regulated HuR-inducible HeLa cells were used to demonstrate that cytoplasmic HuR overexpression promoted a 4-fold increase in exosomes produced. Furthermore, HuR was detected in exosomes produced only from HuR-overexpressing cells. The presence of HuR in exosomes directly impacted mRNA cargo, as selective uptake of ARE-containing mRNAs were observed in exosomes derived from HuR overexpressing cells. To test if these effects were seen in CRC cells that endogenously overexpress HuR, exosome levels from CRC cells were compared to normal primary human intestinal epithelial and myofibroblast cells. When normalized to total cell numbers, CRC cells secrete ~3-fold greater exosome levels than normal cells and enhanced exosome production was dependent upon HuR. Knockdown of HuR in CRC cells directly impacted exosome secretion to levels observed in normal cells. HuR was also only detected in exosomes produced from CRC cells. Using an inducible model of RasV12-mediated transformation of intestinal epithelial cells, endogenous HuR overexpression was observed concomitant with 4-fold greater levels of exosomes containing HuR as cargo. These findings were reflected in vivo where GI-tumor bearing APCMin/+ mice produced ~3-fold more serum exosomes, with HuR as exosomal cargo in APCMin/+ mice whereas no expression of exosomal HuR was detected in wild-type mice. This work has identified a novel connection between HuR-mediated post-transcriptional regulation and tumor-derived exosome production, along with providing the first evidence indicating the presence of exosomal HuR as a serum-based CRC biomarker.Citation Format: Ranjan Preet, Shufei Zhuang, Wei-Ting Hung, Lane K. Christenson, Dan A. Dixon. The RNA binding protein HuR enhances exosome secretion in colorectal cancer. [abstract]. In: Proceedings of the AACR Special Conference: Function of Tumor Microenvironment in Cancer Progression; 2016 Jan 7–10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2016;76(15 Suppl):Abstract nr A41.
Wednesday, July 27, 2016 11:11 AM|MONTAGNANI, F., DI LEONARDO, G., PINO, M. S., MARTELLA, F., PERBONI, S., RIBECCO, A., FIORETTO, L.|Anticancer Research recent issues|Labels: CRC

Background: It is not clear if progression-free survival (PFS) is a good surrogate end-point for overall survival (OS) for metastatic colorectal cancer if antiangiogenic therapies are used. Materials and Methods: We investigated randomized controlled trials testing antiangiogenic agents against chemotherapy. Log hazard ratios (HR) for PFS and OS were used to construct linear regression models. The surrogate threshold effect (STE) was calculated. Results: Thirteen studies and 24 comparison arms were available, including 7,179 patients. This model returned a significant correlation between PFS and OS (R2=0.68, p<0.001) with an STE of 0.83. Analysis restricted to first-line gave similar results (R2=0.68, p<0.001, STE=0.75). Conclusion: There is a significant correlation between the effect of treatment on PFS and OS. PFS remains a good surrogate end-point for OS even if anti-angiogenic agents are used.

Wednesday, July 27, 2016 11:11 AM|PLOQUIN, A., ZERIMECH, F., ESCANDE, F., ADENIS, A., GIRAUD, C., GASNAULT, L., BOURGEOIS, V., DESAUW, C., HEBBAR, M.|Anticancer Research recent issues|Labels: RAS, CRC

Background/Aim: We previously identified three clinical predictive factors of efficacy of cetuximab-irinotecan. Here, we analyzed the clinical characteristics of patients with metastatic colorectal cancer (CRC) in order to detect potent correlations with KRAS mutations. Patients and Methods: We conducted a retrospective, multicenter study between 2008 and 2012. We included patients with metastatic colorectal adenocarcinomas, previously treated by irinotecan, and with an available KRAS mutation test. Results: We included 299 patients. The median age was 60 years; the median number of metastatic sites was 2. One hundred and eight patients (36.1%) had a previous objective response to irinotecan. The median interval between diagnosis and irinotecan discontinuation was 1.94 years. A KRAS mutation was detected in 133 patients (44.5%). In univariate and multivariate analyses, none of the assessed factors was associated with the presence of a KRAS mutation. Conclusion: No easily clinically assessable parameter was significantly associated with KRAS mutations in patients with colorectal cancer.

Wednesday, July 27, 2016 11:11 AM|SUEDA, T., SAKAI, D., KUDO, T., SUGIURA, T., TAKAHASHI, H., HARAGUCHI, N., NISHIMURA, J., HATA, T., HAYASHI, T., MIZUSHIMA, T., DOKI, Y., MORI, M., SATOH, T.|Anticancer Research recent issues|Labels: CRC

Background: Regorafenib and TAS-102 are novel antitumor agents for patients with metastatic colorectal cancer (mCRC) whose disease has progressed after standard therapies. In randomized trials, regorafenib and TAS-102 prolonged survival in patients with mCRC. However, the appropriate selection of regorafenib or TAS-102 in treatment strategy has not yet been established. Patients and Methods: We performed a retrospective analysis, between March 2013 and July 2015, on the efficacy and safety of regorafenib or TAS-102. Results: Thirty-seven patients with mCRC treated with regorafenib or TAS-102 were included. Of these 37 patients, 23 first received regorafenib and 14 received TAS-102. The median progression-free survival and overall survival were 3.0 and 5.8 months, respectively, in the regorafenib group and 2.1 and 6.3 months, respectively, in the TAS-102 group. Drug-related adverse events (AEs) and grade ≥3 AEs were 23 (100%) and 10 (43.5%), respectively, in the regorafenib group and 13 (92.9%) and 2 (14.3%), respectively, in the TAS-102 group. The most frequent grade ≥3 AEs were hepatotoxicity (17.4%) and hand-foot syndrome (13.0%) in the regorafenib group, and neutropenia (14.3%) in the TAS-102 group. In subgroup analysis, the median overall survival was 11.5 months in patients receiving crossover treatment with regorafenib to TAS-102, and 7.6 months in those receiving crossover treatment with TAS-102 to regorafenib. Conclusion: Our results showed that regorafenib and TAS-102 have comparable efficacy but different toxicity profiles in patients with mCRC. Both are considered new salvage treatment options. Differences in the toxicity profiles between the two treatments will help in choosing regorafenib or TAS-102.

Wednesday, July 27, 2016 11:11 AM|ILHAN, N., GUNGOR, H., GUL, H. F., EROKSUZ, H.|Anticancer Research recent issues|Labels: TGF, VEGF, CRC

Background/Aim: Endoglin (CD105) is a receptor for the transforming growth factor-beta 1 (TGFβ1) with crucial role in vascular development and angiogenesis. Additionally, vascular endothelial growth factor (VEGF) overexpression has been associated with advanced stage and poor survival for several cancer types. These molecules have been shown to be useful markers for identifying proliferating endothelium involved in tumor angiogenesis, especially in patients with cancer at risk of developing metastases. The aim of this study was to evaluate the relationship between VEGF and endoglin expression in an experimental model of colorectal cancer, as well as to investigate the effect of cyclooxygenase-2 (COX2) inhibitors on tumor development incidence. Materials and Methods: Colon cancer was induced with 1,2-dimethylhydrazine dihydrochloride (DMH). Celecoxib and diclofenac treatment was started simultaneously with DMH induction. Endoglin protein expression was performed using western blot analysis. VEGF plasma concentrations were measured by enzyme-linked immunosorbent assay. Results: In histopathological evaluations, no pathological change was observed in control rats, while adenocarcinoma (62.5%), dysplasia (31.25%) and inflammation (6.25%) were detected in the group given DMH. In treatment groups, a marked decrease was observed in adenocarcinoma rate. Expression of endoglin protein was significantly elevated in the DMH group compared to controls (p<0.001). No statistically significant difference was noted between treatment groups and DMH group regarding endoglin expression but a decrease was detected in the celecoxib-treated groups. Conclusion: It was confirmed by histopathology and western blotting that COX2 inhibitors, particularly celecoxib, decrease the rate of disease and slow-down progression of existing CRC. These data show that endoglin expression may have an important role in tumor angiogenesis and predict of tumor invasion.

Wednesday, July 27, 2016 11:11 AM|BABA, H., ISHIKAWA, T., MOGUSHI, K., ISHIGURO, M., UETAKE, H., TANAKA, H., SUGIHARA, K.|Anticancer Research recent issues|Labels: CRC, biomarker diagnostic

Background/Aim: The integration of gene expression analysis and genomic profiling represents an efficient approach to the discovery of cancer-related genes. Lymph node metastasis (LNM) is a significant prognostic factor in colorectal cancer (CRC). Detection and analysis of factors related to LNM will help develop new diagnostic methods or therapies. In this study, we aimed to identify genes that are significantly related to LNM in CRC through integrated copy number analysis (CNA) and gene expression analysis. Materials and Methods: Genes showing both up-regulated expression and copy number gains in cases involving CRC with LNM were extracted as candidate biomarkers. Expression of the mRNA of the final candidate was validated in 124 patients using quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays. Expression of the protein encoded by this candidate gene was assessed using immunohistochemical (IHC) staining of tissue from 328 patients. The association between protein expression and clinicopathological features was also examined. Results: Special AT-rich sequence-binding protein 1 (SATB1) was extracted from integrated microarray analysis. SATB1 mRNA expression in cancer tissue was significantly higher in patients with LNM than without LNM. SATB1 protein overexpression was significantly associated with LNM. Moreover, overexpression of SATB1 was an independent poor prognostic factor in stage I-III, especially in stage II CRC. Conclusion: SATB1 may play an important role in LNM of CRC. SATB1 may be a biomarker of LNM and of recurrence after surgery for CRC.

Monday, July 18, 2016 3:00 PM|Bioorganic and Medicinal Chemistry Letters|Bioorganic and Medicinal Chemistry Letters via|Comments|Labels: caspase, CRC
Authors: Shin SY, Ahn S, Yoon H, Jung H, Jung Y, Koh D, Lee YH, Lim Y Abstract To develop potent chemotherapeutic agents for treating colorectal cancers, polymethoxylated 3-naphthyl-5-phenylpyrazoline-carbothioamide derivatives were designed. Twenty-two novel derivatives were synthesized and their cytotoxicities were measured using a clonogenic long-term survival assay. Of these derivatives, 3-(1-hydroxynaphthalen-2-yl)-N-(3-methoxyphenyl)-5-(4-methoxyphenyl)-pyrazoline-1-carbothioamide (NPC 15) exhibited the best half-maximal cell growth inhibitory concentrations (196.35nM). To explain its cytotoxicity, further biological experiments were performed. Treatment with NPC 15 inhibited cell cycle progression and triggered apoptosis through the caspase-mediated pathway. Its inhibitory e...
Friday, July 15, 2016 8:31 AM|Onclive Colorectal Cancer Articles|Labels: CRC, biomarker diagnostic
A liquid biopsy test of circulating tumor DNA for residual disease may be able to determine the risk of recurrence for patients with stage II colon cancer beyond radiologic tests and clinicopathological risk features.
Thursday, July 14, 2016 10:05 PM|Ghosh, P., Tie, J., Muranyi, A., Singh, S., Brunhoeber, P., Leith, K., Bowermaster, R., Liao, Z., Zhu, Y., LaFleur, B., Tran, B., Desai, J., Jones, I., Croxford, M., Jover, R., Goel, A., Waring, P., Hu, S., Teichgraber, V., Rohr, U.-P., Ridder, R., Shanmugam, K., Gibbs, P.|Clinical Cancer Research recent issues|Labels: CRC, biomarker diagnostic

Purpose: Prognostic markers that identify patients with stage II colon cancers who are at the risk of recurrence are essential to personalize therapy. We evaluated the potential of GIV/Girdin as a predictor of recurrence risk in such patients.

Experimental Design: Expression of full-length GIV was evaluated by IHC using a newly developed mAb together with a mismatch repair (MMR)-specific antibody panel in three stage II colon cancer patient cohorts, that is, a training (n = 192), test (n = 317), and validation (n = 181) cohort, with clinical follow-up data. Recurrence risk stratification models were established in the training cohort of T3, proficient MMR (pMMR) patients without chemotherapy and subsequently validated.

Results: For T3 pMMR tumors, GIV expression and the presence of lymphovascular invasion (LVI) were the only factors predicting recurrence in both training (GIV: HR, 2.78, P = 0.013; LVI: HR, 2.54, P = 0.025) and combined test and validation (pooled) cohorts (GIV: HR, 1.85, P = 0.019; LVI: HR, 2.52, P = 0.0004). A risk model based on GIV expression and LVI status classified patients into high- or low-risk groups; 3-year recurrence-free survival was significantly lower in the high-risk versus low-risk group across all cohorts [Training: 52.3% vs. 84.8%; HR, 3.74, 95% confidence interval (CI), 1.50–9.32; Test: 85.9% vs. 97.9%, HR, 7.83, 95% CI, 1.03–59.54; validation: 59.4% vs. 84.4%, HR, 3.71, 95% CI, 1.24–11.12].

Conclusions: GIV expression status predicts recurrence risk in patients with T3 pMMR stage II colon cancer. A risk model combining GIV expression and LVI status information further enhances prediction of recurrence. Further validation studies are warranted before GIV status can be routinely included in patient management algorithms. Clin Cancer Res; 22(14); 3488–98. ©2016 AACR.

Thursday, July 14, 2016 3:00 PM|Cancer|Cancer via|Comments|Labels: CRC
CONCLUSIONSIn what to the authors' knowledge is the largest group of patients with stage II colon cancer evaluated to date, improved OS was found to be associated with adjuvant chemotherapy regardless of treatment regimen, patient age, or high‐risk pathologic risk features. Cancer 2016. © 2016 American Cancer Society. (Source: Cancer)
CONCLUSIONSProviding FIT kits and educational print materials to black individuals in community settings resulted in high rates of CRC screening. The study also identified subgroups of participants who were less likely to return an FIT kit and provides insight for future interventions. Cancer 2016. © 2016 American Cancer Society. (Source: Cancer)
Tuesday, July 12, 2016 10:19 AM|Onclive Colorectal Cancer Articles|Labels: RAS, CRC
The debate between the phase III CALGB/SWOG 80405 and FIRE-3 studies has been settled, as findings of a retrospective analysis of 80405 show that tumor location is significant in determining survival outcomes for patients with KRAS wild-type metastatic colorectal cancer.
Monday, July 4, 2016 12:51 AM|News-Medical.Net Interleukin 2 News Feed|Comments|Labels: CRC, clinical trial, IL
A novel anti-interleukin 1-alpha antibody has shown a significant impact on symptoms, and a high level of safety and tolerability in patients with advanced colorectal cancer, according to phase III data presented at the European Society for Medical Oncology's 18th World Congress of Gastrointestinal Cancer in Barcelona, Spain.
Thursday, June 30, 2016 10:05 PM|Montero, A. J., Kwon, D., Flores, A., Kovacs, K., Trent, J. C., Benedetto, P., Rocha-Lima, C., Merchan, J. R.|Clinical Cancer Research recent issues|Labels: CRC, clinical trial

Purpose: To evaluate the safety, MTD, pharmacokinetics/pharmacodynamics, and early clinical activity of ixabepilone given either weekly or every 3 weeks in combination with daily sunitinib in patients with advanced solid tumors.

Experimental Design: Eligible patients received either weekly (schedule A) or every 3 weeks (schedule B) ixabepilone at escalating doses (schedule A: 7.5, 15, or 20 mg/m2; schedule B: 20, 30, or 40 mg/m2), and oral sunitinib (37.5 mg daily), starting on day 8 of cycle 1. Dose-limiting toxicities (DLT) were assessed during cycle 1.

Results: The ixabepilone and sunitinib combination was fairly well tolerated. DLTs were observed in 3 subjects (1 in schedule 3A and 2 in schedule 3B). The most common grade 3–4 hematologic and nonhematologic adverse events were leukopenia and fatigue, respectively. Four patients (3 in schedule A) achieved a partial response, while 13 patients had stable disease. Nine of 17 heavily pretreated colorectal cancer patients had clinical benefit. Coadministration of sunitinib with ixabepilone on a weekly (but not every 3 week) schedule was associated with a significant increase in the half-life and a significant decrease in the clearance of ixabepilone. Correlative studies demonstrated a significant association between higher baseline plasma angiogenic activity (PAA) and clinical benefit in schedule A patients. Weekly, but not every 3 weeks, ixabepilone led to a significant decrease in PAA postbaseline.

Conclusions: Coadministration of ixabepilone with sunitinib has acceptable toxicity and encouraging clinical activity in heavily pretreated patients, particularly in patients with metastatic colorectal cancer. Clin Cancer Res; 22(13); 3209–17. ©2016 AACR.

Thursday, June 30, 2016 10:05 PM|Morita, R., Hirohashi, Y., Torigoe, T., Ito-Inoda, S., Takahashi, A., Mariya, T., Asanuma, H., Tamura, Y., Tsukahara, T., Kanaseki, T., Kubo, T., Kutomi, G., Mizuguchi, T., Terui, T., Ishitani, K., Hashino, S., Kondo, T., Minagawa, N., Takahashi, N., Taketomi, A., Todo, S., Asaka, M., Sato, N.|Clinical Cancer Research recent issues|Labels: CRC

Purpose: Cancer-initiating cells (CICs) are thought to be essential for tumor maintenance, recurrence, and distant metastasis, and they are therefore reasonable targets for cancer therapy. Cancer immunotherapy is a novel approach to target cancer. In this study, we aimed to establish novel CIC-targeting immunotherapy.

Experimental Design: Colorectal cancer (CRC) CICs were isolated as side population (SP) cells. The gene expression profile of CRC CICs was analyzed by cDNA microarray and RT-PCR. Protein expression of olfactory receptor family 7 subfamily C member 1 (OR7C1) were analyzed by Western blot and immunohistochemical staining. The functions of OR7C1 were analyzed by gene overexpression and gene knockdown using siRNAs. OR7C1-positive cells were isolated by a flow cytometer and analyzed. CTLs specific for OR7C1 peptide were generated, and the antitumor effect was addressed by mice adoptive transfer model.

Results: OR7C1 has essential roles in the maintenance of colon CICs, and the OR7C1-positive population showed higher tumorigenicity than that of the OR7C1-negative population, indicating that OR7C1 is a novel functional marker for colon CIC. Immunohistochemical staining revealed that OR7C1 high expression was correlated with poorer prognosis in CRC patients. OR7C1-derived antigenic peptide-specific CTLs showed specific cytotoxicity for CICs, and an OR7C1-specific CTL clone showed a greater antitumor effect than did a CTL clone targeting all cancer cells in a CTL adoptive transfer mouse model.

Conclusions: OR7C1 is a novel marker for colon CICs and can be a target of potent CIC-targeting immunotherapy. Clin Cancer Res; 22(13); 3298–309. ©2016 AACR.

Thursday, June 30, 2016 10:05 PM|Matsusaka, S., Hanna, D. L., Cao, S., Zhang, W., Yang, D., Ning, Y., Sunakawa, Y., Okazaki, S., Berger, M. D., Miyamato, Y., Parekh, A., Stintzing, S., Loupakis, F., Lenz, H.-J.|Clinical Cancer Research recent issues|Labels: VEGF, CRC, clinical trial, IL

Purpose: The IL6/STAT3 axis promotes inflammation, angiogenesis, and cancer. The effect of genetic variants within this pathway on benefit from antiangiogenic cancer therapy is unknown. We tested whether SNPs in genes involved in IL6/STAT3 signaling can predict efficacy of bevacizumab-based chemotherapy in metastatic colorectal cancer (mCRC) patients.

Experimental Design: Associations between potentially functional IL6 (rs2069837 and rs1800795) and STAT3 (rs744166 and rs4796793) SNPs and clinical outcomes [progression-free survival (PFS), overall survival, and tumor response rate] were evaluated in mCRC patients receiving first-line FOLFIRI plus bevacizumab in two randomized phase III trials: TRIBE (n = 223, training cohort) and FIRE-3 (n = 288, validation cohort). Patients receiving FOLFIRI plus cetuximab in FIRE-3 (n = 264) served as a control cohort. The interaction between genotype and primary tumor location with clinical outcomes was examined. Genomic DNA isolated from whole blood or tumor tissue was analyzed by PCR-based direct sequencing.

Results: Patients with an IL6 rs2069837 G allele treated with FOLFIRI plus bevacizumab had an inferior PFS than those with the A/A genotype in TRIBE [9.4 vs. 11.1 months; HR = 1.53; 95% confidence interval (CI), 1.12–2.10; P = 0.004] and FIRE-3 (8.8 vs. 10.9 months; HR = 1.40; 95% CI, 1.06–1.85; P = 0.015). These associations were confirmed in multivariable analyses and were not seen in the control cohort. In subgroup analysis, the effect of IL6 rs2069837 on PFS was present only in patients with left-sided cancers, but the test for interaction was not significant.

Conclusions: IL6 rs2069837 genotype is a clinically relevant prognostic factor in mCRC patients treated with first-line bevacizumab-based chemotherapy. Clin Cancer Res; 22(13); 3218–26. ©2016 AACR.

Thursday, June 30, 2016 10:05 PM|Unknown Author|Cancer Discovery recent issues|Labels: CRC

Elevated FOXP3, in the absence of FOXP3lo T cells, is associated with poor colon cancer prognosis.

Thursday, June 30, 2016 10:05 PM|Unknown Author|Cancer Discovery recent issues|Labels: EGFR, CRC

According to data from a phase II study, the anti-HER2 combination of trastuzumab and lapatinib is active in patients with KRAS–wild-type, HER2-positive metastatic colorectal cancer. None of the study participants responded to previous, standard treatment with the EGFR inhibitors cetuximab or panitumumab, but 30% achieved an objective response to dual HER2 blockade.

Thursday, June 30, 2016 10:05 PM|Jacobs, E. T., Haussler, M. R., Alberts, D. S., Kohler, L. N., Lance, P., Martinez, M. E., Roe, D. J., Jurutka, P. W.|Cancer Prevention Research recent issues|Labels: CRC

Although hydrophobic bile acids have been demonstrated to exhibit cytotoxic and carcinogenic effects in the colorectum, ursodeoxycholic acid (UDCA) has been investigated as a potential chemopreventive agent. Vitamin D has been shown to play a role in both bile acid metabolism and in the development of colorectal neoplasia. Using a cross-sectional design, we sought to determine whether baseline circulating concentrations of the vitamin D metabolites 25(OH)D and 1,25(OH)2D were associated with baseline fecal bile acid concentrations in a trial of UDCA for the prevention of colorectal adenoma recurrence. We also prospectively evaluated whether vitamin D metabolite concentrations modified the effect of UDCA on adenoma recurrence. After adjustment for age, sex, BMI, physical activity, and calcium intake, adequate concentrations of 25(OH)D (≥30 ng/mL) were statistically significantly associated with reduced odds for high levels of total [OR, 0.61; 95% confidence interval (CI), 0.38–0.97], and primary (OR, 0.61; 95% CI, 0.38–0.96) bile acids, as well as individually with chenodeoxycholic acid (OR, 0.39; 95% CI, 0.24–0.63) and cholic acid (OR, 0.56; 95% CI, 0.36–0.90). No significant associations were observed for 1,25(OH)2D and high versus low fecal bile acid concentrations. In addition, neither 25(OH)D nor 1,25(OH)2D modified the effect of UDCA on colorectal adenoma recurrence. In conclusion, this is the first study to demonstrate an inverse relationship between circulating levels of 25(OH)D and primary fecal bile acid concentrations. These results support prior data demonstrating that vitamin D plays a key role in bile acid metabolism, and suggest a potential mechanism of action for 25(OH)D in colorectal cancer prevention. Cancer Prev Res; 9(7); 589–97. ©2016 AACR.

Thursday, June 30, 2016 10:05 PM|Sheaffer, K. L., Elliott, E. N., Kaestner, K. H.|Cancer Prevention Research recent issues|Labels: CRC

Intestinal cancer is a heterogeneous disease driven by genetic mutations and epigenetic changes. Approximately 80% of sporadic colorectal cancers are initiated by mutation and inactivation of the adenomatous polyposis coli (APC) gene, which results in unrestrained intestinal epithelial growth and formation of adenomas. Aberrant DNA methylation promotes cancer progression by the inactivation of tumor suppressor genes via promoter methylation. In addition, global DNA hypomethylation is often seen before the formation of adenomas, suggesting that it contributes to neoplastic transformation. Previous studies employed mice with a hypomorphic mutation in DNA methyltransferase 1 (Dnmt1), which exhibited constitutive global DNA hypomethylation and decreased tumorigenesis in the ApcMin/+ mouse model of intestinal cancer. However, the consequences of intestinal epithelial-specific acute hypomethylation during ApcMin/+ tumor initiation have not been reported. Using temporally controlled intestinal epithelial-specific gene ablation, we show that total loss of Dnmt1 in the ApcMin/+ mouse model of intestinal cancer causes accelerated adenoma initiation. Deletion of Dnmt1 precipitates an acute response characterized by hypomethylation of repetitive elements and genomic instability, which surprisingly is followed by remethylation with time. Two months post-Dnmt1 ablation, mice display increased macroadenoma load, consistent with a role for Dnmt1 and DNA methylation in maintaining genomic stability. These data suggest that DNA hypomethylation plays a previously unappreciated role in intestinal adenoma initiation. Cancer Prev Res; 9(7); 534–46. ©2016 AACR.

See related article by Lee and Laird, p. 509

Thursday, June 30, 2016 10:05 PM|Dunne, P. D., McArt, D. G., O'Reilly, P. G., Coleman, H. G., Allen, W. L., Loughrey, M., Van Schaeybroeck, S., McDade, S., Salto-Tellez, M., Longley, D. B., Lawler, M., Johnston, P. G.|Cancer Immunology Research recent issues|Labels: PD-1/PD-L1, CRC

A recent phase II study of patients with metastatic colorectal carcinoma showed that mismatch repair gene status was predictive of clinical response to PD-1–targeting immune checkpoint blockade. Further examination revealed strong correlation between PD-L1 protein expression and microsatellite instability (MSI) in stage IV colorectal carcinoma, suggesting that the amount of PD-L1 protein expression could identify late-stage patients who might benefit from immunotherapy. To assess whether the clinical associations between PD-L1 gene expression and MSI identified in metastatic colorectal carcinoma are also present in stage II/III colorectal carcinoma, we used in silico analysis to elucidate the cell types expressing the PD-L1 gene. We found a statistically significant association of PD-L1 gene expression with MSI in early-stage colorectal carcinoma (P < 0.001) and show that, unlike in non–colorectal carcinoma tumors, PD-L1 is derived predominantly from the immune infiltrate. We demonstrate that PD-L1 gene expression has positive prognostic value in the adjuvant disease setting (PD-L1low vs. PD-L1high HR = 9.09; CI, 2.11–39.10). PD-L1 gene expression had predictive value, as patients with high PD-L1 expression appear to be harmed by standard-of-care treatment (HR = 4.95; CI, 1.10–22.35). Building on the promising results from the metastatic colorectal carcinoma PD-1–targeting trial, we provide compelling evidence that patients with PD-L1high/MSI/immunehigh stage II/III colorectal carcinoma should not receive standard chemotherapy. This conclusion supports the rationale to clinically evaluate this patient subgroup for PD-1 blockade treatment. Cancer Immunol Res; 4(7); 582–91. ©2016 AACR.

Thursday, June 30, 2016 10:05 PM|Tada, K., Kitano, S., Shoji, H., Nishimura, T., Shimada, Y., Nagashima, K., Aoki, K., Hiraoka, N., Honma, Y., Iwasa, S., Okita, N., Takashima, A., Kato, K., Yamada, Y., Katayama, N., Boku, N., Heike, Y., Hamaguchi, T.|Cancer Immunology Research recent issues|Labels: CRC

It remains unclear whether the immunologic status of cells in peripheral blood can be used as a prognostic indicator of response to treatment for patients with unresectable metastatic colorectal cancer (MCRC). We therefore investigated the relationship between the pretreatment immunologic status of 40 patients with MCRC who planned to receive the first-line chemotherapy and their progression-free survival. Twenty-five immune cell subsets, including monocytic myeloid-derived suppressor cells (M-MDSC) and effector memory T cells (TEM), were measured by multicolor-flow cytometry. We divided patients into high and low (above and below the median, respectively) groups based on the median value for each immune cell subset and compared progression-free survival of the two groups. Patients with high M-MDSC, low CD4+ TEM, or low CD8+ TEM quantities had significantly shorter progression-free survival (P = 0.004, 0.005, and 0.002, respectively). Patients were classified into two prognostic groups based on numbers of adverse factors; having two or three adverse factors (n = 21, 52.5%) was correlated with significantly shorter progression-free survival compared with none or one (n = 19, 47.5%; P < 0.001). The presence of two or three adverse factors was an independent poor prognostic factor for progression-free survival (HR, 9.2; 95% confidence interval, 2.5–34.2; P < 0.001). These results provide evidence that pretreatment peripheral immune status can inform the outcome of patients with MCRC treated with first-line chemotherapy. Cancer Immunol Res; 4(7); 592–9. ©2016 AACR.

Monday, June 27, 2016 10:52 AM|MUKOHYAMA, J., SHIMONO, Y., YAMASHITA, K., SUMI, Y., MUKOHARA, T., MINAMI, H., KAKEJI, Y.|Anticancer Research recent issues|Labels: CRC

Background: Cancer stem cells (CSCs) have a high tumorigenic ability to form patient-derived tumor xenografts (PDXs). PDXs are an attractive pre-clinical model, but gene expression and biological behavior of cancer cells in the tumor will change during establishment and passage of PDXs. Materials and Methods: Human colon cancer PDX was established and passaged either subcutaneously or orthotopically into the murine intestine. Histology and flow cytometric profile of the surgical specimen and the PDX were analyzed. CSCs were then isolated from the tumors and their microRNA (miRNA) expression was analyzed by semi-quantitative polymerase chain reaction. Results: The surgical specimens and PDXs were histologically similar. The size of CSC population increased and expression of miRNAs in CSCs changed in the passaged PDXs. Expression of oncogenic miRNAs was highly up-regulated in the CSCs of the orthotopically passaged PDXs. Conclusion: The xenotransplantation site and the number of tumor passages affect the miRNA expression of human colon CSCs.

Monday, June 27, 2016 10:52 AM|SHIRAIWA, S., KINUGASA, T., KAWAHARA, A., MIZOBE, T., OHCHI, T., YUGE, K., FUJINO, S., KATAGIRI, M., SHIMOMURA, S., TAJIRI, K., SUDO, T., KAGE, M., KUWANO, M., AKAGI, Y.|Anticancer Research recent issues|Labels: EGFR, CRC

Background/aim: Y-Box-binding protein-1 (YB-1), a DNA/RNA-binding protein, is an important oncogenic transcription and translation factor. We aimed to evaluate the relationships between nuclear YB-1 expression, epidermal growth factor receptor (EGFR) status, and poor clinical outcomes in patients with colorectal cancer (CRC). Materials and Methods: Nuclear YB-1 expression was immunohistochemically analyzed in CRC tissues obtained from 124 patients who underwent curative resection between 2005 and 2008. Correlations between nuclear YB-1 expression, various clinicopathological characteristics, EGFR status, and prognostic factors were evaluated. Results: High-grade nuclear YB-1 expression was detected in 62.9% of cases and was found to be an independent predictor of poorer overall survival (p<0.001) and relapse-free survival (p<0.001). A trend was also observed towards a positive correlation between nuclear YB-1 expression and EGFR status (p=0.051). Conclusion: Nuclear YB-1 expression is a useful prognostic biomarker that correlates with EGFR status in patients with CRC.

Monday, June 27, 2016 10:52 AM|YOSHIDA, Y., NAITO, M., YAMADA, T., AISU, N., DAIBO, K., MERA, T., TANAKA, T., NAITO, K., YASUMOTO, K., KAMIGAKI, T., GOTO, S., YAMASHITA, Y., HASEGAWA, S.|Anticancer Research recent issues|Labels: CRC

Background/Aim: Adoptive immunotherapy of cancer is evolving with the development of novel technologies that generate proliferation of large numbers of αβ and T cells. We evaluated the safety and efficacy of the combination of adoptive immunotherapy using αβ T cells with chemotherapy for stage IV colorectal cancer (CRC). Patients and Methods: Fifteen patients with advanced or recurrent CRC received XELOX + bevacizumab + ex vivo expanded αβ T lymphocytes as a first-line chemoimmunotherapy. Results: Median age of the 15 patients (4 men, 11 women) was 65 years (range=49-80). Median progression-free survival was 21.3 months. Response rate was 80% (complete response (CR)=26.7%, partial response (PR)=53.3%, stable disease (SD)=20% and progressive disease (PD)=0%). Most adverse events were mild to moderate regarding their intensity and immunotherapy-associated toxicity was minimal. Conclusion: Combination of adoptive αβ T cell immunotherapy with chemotherapy for stage IV CRC is feasible and safe.

Monday, June 27, 2016 10:52 AM|AKINWANDE, O., SCOGGINS, C., MARTIN, R. C. G.|Anticancer Research recent issues|Labels: CRC

Aim: To report our early experience on the feasibility and safety of 70-150 μm drug-eluting beads loaded with irinotecan (M1-DEBIRI) for treating unresectable hepatic colorectal metastases. Patients and Methods: An Institutional Review Board-approved, prospectively maintained, multi-institutional registry was evaluated from 2/2009 to 8/2013. Fifteen consecutive patients presenting with liver-dominant metastatic colorectal cancer were treated with M1-DEBIRI. Kaplan–Meier statistics was used to evaluate hepatic progression-free-survival and overall survival. Results: Fifteen patients underwent 32 DEBIRI treatments. The mean prescribed dose was 100 mg of irinotecan (range=100-200 mg). In 75% of treatments (n=24), 100% of the prescribed dose was delivered before complete stasis. In 97% of treatments (n=31), at least 50% (median 100 mg, range 25-150 mg) of the prescribed dose was delivered. There was grade 2 abdominal pain after one treatment (3%). In another patient, increased total bilirubin (1.1 to 3.1 mg/dl) was seen after one treatment. There was 42% reduction in median carcinoembryonic antigen level and 33% (5/15 patients) with Response Evaluation Criteria in Solid Tumors (RECIST) objective response (complete and partial). Modified RECIST and European Association for the study of the Liver (EASL) objective response rates were both 73% (11/15 patients). The disease control rate was 93% (14/15 patients). Hepatic progression-free-survival and overall survival were 8 and 13 months respectively. Disease in one patient was down-staged to resection (6%). Conclusion: M1-DEBIRI appears to be safe and feasible in the treatment of metastatic colorectal cancer. Smaller beads also provide efficient irinotecan dose delivery. Larger studies are needed to validate these findings.

Monday, June 27, 2016 10:52 AM|MATSUI, T., NAGATA, N., HIRATA, K., OKAZAKI, S., SATO, S., NAKAMURA, M., KIM, H., OBA, K., SAKAMOTO, J., MISHIMA, H.|Anticancer Research recent issues|Labels: VEGF, CRC, clinical trial

Aim: This phase II study assessed the efficacy and toxicity of an intermittent weekly capecitabine regimen in combination with oxaliplatin (XELOX) plus bevacizumab as a first-line treatment of metastatic colorectal cancer (mCRC). Patients and Methods: Patients with measurable mCRC who were to receive first-line chemotherapy were enrolled onto this disease-oriented multicenter phase II trial. Patients with mCRC were required to have Eastern Cooperative Oncology Group performance status of 0 to 1, to be aged >20 years, and to have adequate organ function. Localization of tumor, toxicities, response rate, progression-free survival (PFS) and time to progression (TTP) were studied. Capecitabine dose was 2,500 mg/m2/day on days 1-7 (n=47) and was increased to 3,000 mg/m2/day (n=5) in combination with oxaliplatin (85 mg/m2) and bevacizumab (5 mg/kg), repeated every 2 weeks. Results: A total of 51 patients were enrolled from 14 institutions from December 2011 to July 2012. The median age was 66 (range=38-85) years, 29 (56.9%) had colonic cancer and 22 (43.1%) had rectal cancer in this study. Pertinent grade 3/4 toxicities were neutropenia (13.7%), peripheral neuropathy (13.7%), hypertension (13.7%), gastrointestinal perforation (3.9%), and hand-foot syndrome (5.9%). The response rate was 51% (one complete and 25 partial responses). Median PFS and TTP were 344 days and 196 days, respectively. Median overall survival has not been reached yet. Conclusion: The first-line treatment of mCRC using a biweekly combination of XELOX plus bevacizumab can also be administered safely and effectively in Japan. It is suggested that this regimen is an appropriate option for the treatment of mCRC.

Monday, June 27, 2016 10:52 AM|KANG, B. W., JEON, H.-S., CHAE, Y. S., LEE, S. J., PARK, J. S., CHOI, G. S., KIM, J. G.|Anticancer Research recent issues|Labels: CRC

The present study analyzed single nucleotide polymorphisms (SNPs) located at putative microRNA(miRNA)-binding sites of the 3’-untranslated region (UTR) in different genes and investigated their impact on the susceptibility to colorectal cancer (CRC). Ninety-two SNPs were selected using an in silico analysis of 3’-UTR SNPs in an SNP database and their miRNA binding efficiency was calculated using several miRNA databases and the HapMap database. Two independent study sets were used: 380 healthy controls and 371 patients with colorectal adenocarcinoma for the discovery set, and 521 healthy controls and 524 patients with colorectal adenocarcinoma for the validation set. The SNP genotyping was performed using a Sequenom MassARRAY. In addition, a luciferase assay was used to investigate whether miR-370 modulated docking protein 3 (DOK3) gene expression when rs2279398G>A was included in the DOK3 3’-UTR region. For the discovery set, 16 out of 92 SNPs were significantly associated with the risk of CRC in at least one of the genetic models. The validation set showed that among these 16 SNPs, DOK3 rs2279398G>A was significantly associated with reduced risk of CRC in a recessive model [adjusted odds ratio (aOR)=0.65, 95% confidence interval (CI)=0.44-0.97, p=0.03]. In a combined analysis, DOK3 rs2279398G>A was associated with a significantly reduced risk of CRC in a co-dominant and recessive model (aOR=0.84, 95% CI=0.73-0.96, p=0.012; aOR=0.65, CI=0.49-0.88, p=0.004, respectively). Significantly lower Renilla activity was also observed with the rs2279398 AA construct when compared to the rs2279398 GG construct (p<0.001). DOK3 rs2279398G>A may affect the expression of DOK3 by altering the miRNA binding efficiency at the miRNA-binding sites of the 3’-UTR in DOK3, thereby impacting CRC tumorigenesis.

Monday, June 27, 2016 10:52 AM|YAMAGUCHI, T., IWASA, S., NAGASHIMA, K., IKEZAWA, N., HAMAGUCHI, T., SHOJI, H., HONMA, Y., TAKASHIMA, A., OKITA, N., KATO, K., YAMADA, Y., SHIMADA, Y.|Anticancer Research recent issues|Labels: RAS, CRC

Background/Aim: Panitumumab and cetuximab are known to be effective treatments for KRAS wild-type metastatic colorectal cancer (mCRC). However, it remains unclear which of these two biologic agents confers the greatest benefit when combined with irinotecan in patients with KRAS wild-type mCRC previously treated with fluoropyrimidine, oxaliplatin and irinotecan. Patients and Methods: Data, from 139 patients who received panitumumab or cetuximab, in combination with irinotecan, for KRAS wild-type mCRC previously treated with fluoropyrimidine, oxaliplatin and irinotecan were analyzed. The efficacy and safety of panitumumab plus irinotecan was compared to that of cetuximab plus irinotecan. Results: Baseline characteristics of the panitumumab plus irinotecan (n=42) and cetuximab plus irinotecan (n=97) groups were similar. Among patients with measurable lesions, the response rate was 34% in the panitumumab plus irinotecan group and 20% in the cetuximab plus irinotecan group. Median progression-free survival (PFS) was 4.3 and 5.7 months in the panitumumab and cetuximab groups, respectively. Median overall survival was 13.6 months with panitumumab and 11.2 months with cetuximab. Conclusion: Panitumumab plus irinotecan was well-tolerated and displayed a similar level of efficacy to that of cetuximab plus irinotecan.

Wednesday, June 15, 2016 10:05 PM|Mathur, P. S., Gierut, J. J., Guzman, G., Xie, H., Xicola, R. M., Llor, X., Chastkofsky, M. I., Perekatt, A. O., Tyner, A. L.|Molecular Cancer Research recent issues|Labels: Trk, CRC

Disruption of the gene encoding Protein Tyrosine Kinase 6 (Ptk6) delayed differentiation and increased growth in the mouse intestine. However, Ptk6-null mice were also resistant to azoxymethane-induced colon tumorigenesis. To further explore functions of PTK6 in colon cancer, expression of epithelial and mesenchymal markers, as well as proliferation, migration, and xenograft tumor growth, was examined in human colon tumor cell lines with knockdown or overexpression of PTK6. PTK6 protein, transcript, and activation were also examined in a human colon tumor tissue array, using immunohistochemistry and qRT-PCR. Knockdown of PTK6 led to the epithelial–mesenchymal transition (EMT) in SW480 and HCT116 cells, whereas overexpression of PTK6 in SW620 cells restored an epithelial phenotype in a kinase-independent manner. PTK6 knockdown also increased xenograft tumor growth of SW480 cells, suggesting tumor suppressor functions. In clinical specimens, PTK6 expression was highest in normal differentiated epithelial cells and reduced in tumors. In contrast, overexpression of constitutively active PTK6 promoted STAT3 and ERK5 activation in colon cancer cells, and endogenous PTK6 promoted cell survival and oncogenic signaling in response to DNA-damaging treatments. These data indicate that PTK6 has complex, context-specific functions in colon cancer; PTK6 promotes the epithelial phenotype to antagonize the EMT in a kinase-independent manner, whereas activation of PTK6 promotes oncogenic signaling.

Implications: Understanding context-specific functions of PTK6 is important, because although it promotes cell survival and oncogenic signaling after DNA damage, expression of PTK6 in established tumors may maintain the epithelial phenotype, preventing tumor progression. Mol Cancer Res; 14(6); 563–73. ©2016 AACR.

Thursday, June 9, 2016 5:56 AM|German Cancer Research Center|Labels: CRC, clinical trial

Metastasized colorectal cancer is difficult to treat. Scientists at the National Center for Tumor Diseases (NCT) and the Heidelberg University Hospital in collaboration with the German Cancer Research Center (DKFZ) have now discovered that the immune system acts as an accomplice to the metastases. Macrophages, also called scavenger cells, play a vital role in this process. The metastases in the liver influence macrophages in a way that helps tumor cells grow and spread. The reason for this is a signal pathway which is also used by the human immunodeficiency virus (HIV) as an entry point into human cells. One inhibitor is already being used therapeutically in HIV patients. The scientists have now tested this drug in pre-clinical experiments and in a subsequent study involving 14 patients with advanced metastatic colorectal cancer. This Phase I study was funded by the Dietmar-Hopp Foundation and the promising results have just been published in Cancer Cell.The NCT is a joint institution of the German Cancer Research Center (DKFZ), the Heidelberg University Hospital and the German Cancer Aid (Deutsch Krebshilfe).



Tuesday, June 7, 2016 10:41 AM|Kuroiwa-Trzmielina, J., Wang, F., Rapkins, R. W., Ward, R. L., Buchanan, D. D., Win, A. K., Clendenning, M., Rosty, C., Southey, M. C., Winship, I., Hopper, J. L., Jenkins, M. A., Olivier, J., Hawkins, N. J., Hitchins, M. P.|Clinical Cancer Research Online First Articles|Labels: CRC

Purpose: Methylation of the MGMT promoter is the major cause of O6-methylguanine methyltransferase deficiency in cancer and has been associated with the T variant of the promoter-enhancer SNP rs16906252C>T. We sought evidence for an association between the rs16906252C>T genotype and increased risk of developing a subtype of colorectal cancer (CRC) featuring MGMT methylation, mediated by genotype-dependent epigenetic silencing within normal tissues. Experimental Design: By applying a molecular pathological epidemiology case-control study design, associations between rs16906252C>T and risk for CRC overall, and CRC stratified by MGMT methylation status, were estimated using multinomial logistic regression in two independent retrospective series of CRC cases and controls. The test sample comprised 1054 CRC cases and 451 controls from Sydney, Australia. The validation sample comprised 612 CRC cases and 245 controls from the Australasian Colon Cancer Family Registry (ACCFR). To determine if rs16906252C>T was linked to a constitutively altered epigenetic state, quantitative allelic expression and methylation analyses were performed in normal tissues. Results: An association between rs16906252C>T and increased risk of developing MGMT-methylated CRC in the Sydney sample was observed (OR 3.3; 95%CI=2.0-5.3; P<0.0001), which was replicated in the ACCFR sample (OR 4.0; 95%CI=2.4-6.8; P<0.0001). The T allele demonstrated ~2.5-fold reduced transcription in normal colorectal mucosa from cases and controls, and was selectively methylated in a minority of normal cells, indicating rs16906252C>T represents an expression and methylation quantitative trait locus. Conclusions: We provide evidence that rs16906252C>T is associated with elevated risk for MGMT-methylated CRC, likely mediated by constitutive epigenetic repression of the T allele.

Monday, June 6, 2016 9:26 AM|Cancer Therapy Advisor Videos : Recently Posted|Labels: PD-1/PD-L1, CRC
Dr. Johanna Bendell discusses her research at the ASCO 2016 meeting in Chicago.
Monday, June 6, 2016 4:58 AM|NASIR, A., REISING, L. O., NEDDERMAN, D. M., FULFORD, A. D., UHLIK, M. T., BENJAMIN, L. E., SCHADE, A. E., HOLZER, T. R.|Anticancer Research recent issues|Labels: VEGF, CRC

Background: The vascular endothelial growth factor (VEGF) pathway plays an important role in growth and progression of human cancer, including colorectal carcinomas (CRC). The key mediators of VEGF signaling are VEGFR1, VEGFR2, and VEGFR3, part of a family of related receptor tyrosine kinases. The relative expression, activity, or interplay among these receptors may determine the response of CRC patients to anti-angiogenic therapies. Materials and Methods: We developed technically sound immunohistochemical (IHC) assays to quantify VEGFR1, 2 and 3, and using a well-annotated CRC tissue microarray (TMA), we carried out comprehensive comparative evaluation of the three VEGFRs in archival primary CRC tissues (n=84). For each TMA core, tumor cell VEGFR1 expression was reported as H-score (range=0-300); vascular VEGFR2/VEGFR3 expression was manually scored as the number of receptor-positive tumor stromal vessels. Each case was defined as VEGFR1/ VEGFR2/VEGFR3-negative, low, medium or high. Results: Based on the differential expression of the three VEGFRs, eight VEGFR staining profiles were observed: Triple VEGFR positive (n=12, 14%), VEGFR1 predominant (n=17, 20%), VEGFR2 predominant (n=7, 8%), VEGFR3 predominant (n=1, 1%), VEGFR1/2 predominant (n=39, 46%), VEGFR1/3 predominant (n=2, 2%), VEGFR2/3 predominant (n=3, 4%), and triple-VEGFR-negative (n=3, 4%). Conclusion: Herein we demonstrated heterogeneity of expression of VEGFRs in human CRC stromal vessels and tumor cells. The observed VEGFR expression-based subsets of human CRCs may reflect differences in biology of pathologic angiogenesis in primary CRC tissues. Furthermore, the heterogeneity of expression of VEGFRs unraveled in this analysis merits independent validation in larger cohorts of primary and metastatic human CRC tissues and in pertinent experimental models treated with various anti-angiogenic therapies.

Thursday, June 2, 2016 10:05 PM|Unknown Author|Cancer Discovery recent issues|Labels: CRC

CCL5 produced by T cells at invasive margins of colorectal cancer metastases promotes tumor growth.

Thursday, June 2, 2016 9:01 AM|Nagasaka, T., Mishima, H., Sawaki, A., Shimokawa, M., Inukai, M., Shinozaki, K., Tanioka, H., Nasu, J., Nishina, T., Hazama, S., Okajima, M., Yamaguchi, Y.|BMJ Open Protocol|Labels: VEGF, CRC, clinical trial

Results from several randomised trials suggest that the sequential use of cytotoxic agents in patients with metastatic colorectal cancer (mCRC) has the potential to improve overall survival compared with combination chemotherapy. This study is designed to investigate whether sequential treatment with bevacizumab-based first-line treatment with oxaliplatin is superior to combination treatment of mCRC.

Methods and analysis

The C-cubed (C3) study is a two-arm, multicentre, open-label, randomised phase III trial in Japan comparing the efficacy and safety of sequential capecitabine or 5-fluorouracil plus bevacizumab (Cape/5-FU-Bmab) with escalation to capecitabine or 5-fluorouracil plus oxaliplatin plus bevacizumab (CapeOX/mFOLFOX6-Bmab) versus combination CapeOX/mFOLFOX6-Bmab as the first-line treatment of mCRC. In the sequential arm (Arm A: oxaliplatin ‘wait-and-go’), treatment escalation from Cape/5-FU-Bmab to CapeOX/mFOLFOX6-Bmab is recommended in the case of progressive disease. In the combination arm (Arm B: oxaliplatin ‘stop-and-go’), de-escalation from CapeOX/mFOLFOX6-Bmab to Cape/5-FU-Bmab is possible after 12 weeks of treatment. Re-escalation to CapeOX/mFOLFOX6-Bmab after progressive disease is considered only for patients who received de-escalation of oxaliplatin after 12 weeks of treatment not caused by oxaliplatin-associated toxicity. A target sample size of 304 evaluable patients is considered sufficient to validate an expected HR for time to failure of strategy of the sequential approach ‘wait-and-go’ compared to the combination approach ‘stop-and go’ with 80% power and 2-sided 5% α in case of a true HR<0.69.

Ethics and dissemination

This study is conducted according to the standards of Good Clinical Practice and in compliance with the Declaration of Helsinki 2013 and local regulations, and has been submitted and approved by the Ethical Committee of the Non-Profit Organization MINS Institutional Review Board. The protocol and the trial results, even inconclusive, will be presented at international oncology congresses and published in peer-reviewed journals.

Trial registration number

UMIN000015405, Pre-results.

Tuesday, May 31, 2016 10:05 PM|Borras, E., San Lucas, F. A., Chang, K., Zhou, R., Masand, G., Fowler, J., Mork, M. E., You, Y. N., Taggart, M. W., McAllister, F., Jones, D. A., Davies, G. E., Edelmann, W., Ehli, E. A., Lynch, P. M., Hawk, E. T., Capella, G., Scheet, P., Vilar, E.|Cancer Prevention Research recent issues|Labels: WNT, CRC, preclinical

The molecular basis of the adenoma-to-carcinoma transition has been deduced using comparative analysis of genetic alterations observed through the sequential steps of intestinal carcinogenesis. However, comprehensive genomic analyses of adenomas and at-risk mucosa are still lacking. Therefore, our aim was to characterize the genomic landscape of colonic at-risk mucosa and adenomas. We analyzed the mutation profile and copy number changes of 25 adenomas and adjacent mucosa from 12 familial adenomatous polyposis patients using whole-exome sequencing and validated allelic imbalances (AI) in 37 adenomas using SNP arrays. We assessed for evidence of clonality and performed estimations on the proportions of driver and passenger mutations using a systems biology approach. Adenomas had lower mutational rates than did colorectal cancers and showed recurrent alterations in known cancer driver genes (APC, KRAS, FBXW7, TCF7L2) and AIs in chromosomes 5, 7, and 13. Moreover, 80% of adenomas had somatic alterations in WNT pathway genes. Adenomas displayed evidence of multiclonality similar to stage I carcinomas. Strong correlations between mutational rate and patient age were observed in at-risk mucosa and adenomas. Our data indicate that at least 23% of somatic mutations are present in at-risk mucosa prior to adenoma initiation. The genomic profiles of at-risk mucosa and adenomas illustrate the evolution from normal tissue to carcinoma via greater resolution of molecular changes at the inflection point of premalignant lesions. Furthermore, substantial genomic variation exists in at-risk mucosa before adenoma formation, and deregulation of the WNT pathway is required to foster carcinogenesis. Cancer Prev Res; 9(6); 417–27. ©2016 AACR.

Tuesday, May 31, 2016 10:05 PM|Kanth, P., Bronner, M. P., Boucher, K. M., Burt, R. W., Neklason, D. W., Hagedorn, C. H., Delker, D. A.|Cancer Prevention Research recent issues|Labels: CRC

Sessile serrated colon adenoma/polyps (SSA/P) are found during routine screening colonoscopy and may account for 20% to 30% of colon cancers. However, differentiating SSA/Ps from hyperplastic polyps (HP) with little risk of cancer is challenging and complementary molecular markers are needed. In addition, the molecular mechanisms of colon cancer development from SSA/Ps are poorly understood. RNA sequencing (RNA-Seq) was performed on 21 SSA/Ps, 10 HPs, 10 adenomas, 21 uninvolved colon, and 20 control colon specimens. Differential expression and leave-one-out cross-validation methods were used to define a unique gene signature of SSA/Ps. Our SSA/P gene signature was evaluated in colon cancer RNA-Seq data from The Cancer Genome Atlas (TCGA) to identify a subtype of colon cancers that may develop from SSA/Ps. A total of 1,422 differentially expressed genes were found in SSA/Ps relative to controls. Serrated polyposis syndrome (n = 12) and sporadic SSA/Ps (n = 9) exhibited almost complete (96%) gene overlap. A 51-gene panel in SSA/P showed similar expression in a subset of TCGA colon cancers with high microsatellite instability. A smaller 7-gene panel showed high sensitivity and specificity in identifying BRAF-mutant, CpG island methylator phenotype high, and MLH1-silenced colon cancers. We describe a unique gene signature in SSA/Ps that identifies a subset of colon cancers likely to develop through the serrated pathway. These gene panels may be utilized for improved differentiation of SSA/Ps from HPs and provide insights into novel molecular pathways altered in colon cancer arising from the serrated pathway. Cancer Prev Res; 9(6); 456–65. ©2016 AACR.

Tuesday, May 31, 2016 6:00 PM|Simona Rossi|Non-Coding RNA|Labels: CRC, biomarker diagnostic
MiRNAs are suggested as promising cancer biomarkers. They are stable and extractable from a variety of clinical tissue specimens (fresh frozen or formalin fixed paraffin embedded tissue) and a variety of body fluids (e.g., blood, urine, saliva). However, there are several challenges that need to be solved, considering their potential as biomarkers in cancer, such as lack of consistency between biomarker panels in independent studies due to lack of standardized sample handling and processing, use of inconsistent normalization approaches, and differences in patients populations. Focusing on colorectal cancer (CRC), divergent results regarding circulating miRNAs as prognostic or predictive biomarkers are reported in the literature. In the present review, we summarize the current data on circulating miRNAs as prognostic/predictive biomarkers in patients with localized and metastatic CRC (mCRC).
Sunday, May 22, 2016 6:00 PM|Gregory Yochum|Cancers|Labels: WNT, CRC
Mutations in components of the Wnt/β-catenin signaling pathway drive colorectal cancer (CRC) by deregulating expression of downstream target genes including the c-MYC proto-oncogene (MYC). The critical regulatory DNA enhancer elements that control oncogenic MYC expression in CRC have yet to be fully elucidated. In previous reports, we correlated T-cell factor (TCF) and β-catenin binding to the MYC 3′ Wnt responsive DNA element (MYC 3′ WRE) with MYC expression in HCT116 cells. Here we used CRISPR/Cas9 to determine whether this element is a critical driver of MYC. We isolated a clonal population of cells that contained a deletion of a single TCF binding element (TBE) within the MYC 3′ WRE. This deletion reduced TCF/β-catenin binding to this regulatory element and decreased MYC expression. Using RNA-Seq analysis, we found altered expression of genes that regulate metabolic processes, many of which are known MYC target genes. We found that 3′ WRE-Mut cells displayed a reduced proliferative capacity, diminished clonogenic growth, and a decreased potential to form tumors in vivo. These findings indicate that the MYC 3′ WRE is a critical driver of oncogenic MYC expression and suggest that this element may serve as a therapeutic target for CRC.
Friday, May 13, 2016 4:00 AM|News-Medical.Net Cetuximab News Feed|Comments|Labels: CRC
Cancer researchers have identified a marker that shows up in a blood test that determines which patients with colorectal cancer that has spread would benefit from receiving the drug cetuximab.
Tuesday, May 10, 2016 6:00 PM|Avri Ben-Ze’ev|Cancers|Labels: WNT, CRC
The Wnt-β-catenin signaling pathway is highly conserved during evolution and determines normal tissue homeostasis. Hyperactivation of Wnt-β-catenin signaling is a characteristic feature of colorectal cancer (CRC) development. β-catenin is a major transducer of the Wnt signal from the cytoplasm into the nucleus where it acts as a co-transcriptional activator of β-catenin-TCF target genes. β-catenin is also required for linking cadherin type cell-cell adhesion receptors to the cytoskeleton, and consequently Wnt-β-catenin signaling is an attractive system for investigating the role of adhesion-mediated signaling in both normal intestinal tissue homeostasis and CRC development. In this review, we summarize our studies on one Wnt-β-catenin target gene, L1, a member of the immunoglobulin-like cell adhesion transmembrane receptor family. We describe the mechanisms of L1-mediated signaling in CRC cells, its exclusive localization in invasive areas of CRC tissue, and its ability to increase cell motility and confer metastasis to the liver. We discuss the activation (by L1) of genes via an ezrin-NF-κB pathway and the induction of genes also found in the intestinal stem cell signature. By studying L1 (adhesion)-mediated signaling, we expect to learn about mechanisms regulating both normal intestinal homeostasis and CRC development.
Tuesday, May 10, 2016 8:29 AM|Munakata, K., Uemura, M., Tanaka, S., Kawai, K., Kitahara, T., Miyo, M., Kano, Y., Nishikawa, S., Fukusumi, T., Takahashi, Y., Hata, T., Nishimura, J., Takemasa, I., Mizushima, T., Ikenaga, M., Kato, T., Murata, K., Carethers, J. M., Yamamoto, H., Doki, Y., Mori, M.|Clinical Cancer Research Online First Articles|Labels: proteasome, CRC, clinical trial

Purpose:One of the main reasons for cancer treatment resistance is the existence of cancer stem-like cells (CSCs). Here, we elucidated the relationship between low proteasome activity cells (LPACs) and CSCs. Experimental Design:The human CRC cell lines HCT116, SW480, DLD1, and KM12SM were engineered to stably express a green fluorescent molecule fused to the degron of ornithine decarboxylase, resulting in an accumulation of the fluorescence in LPACs. LPACs were isolated by flow cytometry. Treatment resistance (radiation and chemotherapy) and the capacity of LPACs to act as CSCs were analyzed. Microarray analysis was performed to reveal genes related to treatment resistance. The prognostic impact of potent genes was examined in 190 CRC patients. Results:LPACs had a significantly increased capacity for radioresistance and chemoresistance (5-fluorouracil and oxaliplatin), significantly lower reactive oxygen species activity, and significantly increased sphere-formation capacity compared to non-LPACs. The number of cells in the G0/G1 phase was significantly higher among LPACs. Subcutaneous injection of as few as 20 LPACs led to tumor formation in immunologically incompetent mice. Microarray analysis revealed that the expression of EP300 Interacting Inhibitor of Differentiation 3 (EID3) was significantly increased in LPACs. In vitro assay revealed that EID3 positively controlled cell proliferation and treatment resistance. The high expression of EID was an adverse prognostic indicator in patients with CRC (p=0.0400). Conclusions:LPACs have characteristic treatment resistance and act as CSCs in CRC. In addition, EID3 is one of the potential regulators of treatment resistance in CRC and may be a potential therapeutic target.

Thursday, April 28, 2016 9:59 AM|OGAWA, M., ANAN, T., SUZUKI, T., OKUMA, M., ICHIHARA, K., HASEGAWA, T., YOSHIDA, K., YANAGA, K.|Anticancer Research recent issues|Labels: RAS, CRC, clinical trial

Aim: This prospective study was designed to evaluate the tolerability and the efficacy of bi-weekly SOX (S-1 and oxaliplatin)+cetuximab as first-line chemotherapy for wild-type K-RAS metastatic colorectal cancer. Patients and Methods: We studied patients with previously untreated, unresectable, advanced or recurrent colorectal cancer who were treated in our hospital between October 2010 and March 2013. Their performance status (PS) was 0 to 1. Cetuximab was combined with S-1 and oxaliplatin (SOX+cetuximab). S-1 was given orally at a dose of 40 mg/m2 (40-60 mg, calculated according to body surface area) twice daily after meals for 2 weeks, followed by a 2-week rest (course 1). Oxaliplatin (85 mg/m2) was given on days 1 and 15 of each course. Cetuximab was administered on days 1 (400 mg/m2), 8 (250 mg/m2) and 15 (500 mg/m2) of course 1, followed by every 2 weeks (500 mg/m2) thereafter. Results: The study group comprised of 18 patients. The mean age was 61 (range=32-72) years, the male:female ratio was 10:8 and the PS was 0 in 12 patients and 1 in 6 patients. The median number of administered courses was 6 (range=2-12). The treatment response was complete response (CR) in 2 and partial response (PR) in 10 (response rate=67% (12/18 patients)). The minimum number of treatment courses until a PR was 2, indicating an early response. Liver resection was performed in 4 patients (22.2%). The incidence of any adverse events (Grade 3/4) was 28% (5/18), including skin disorder (16.7%) as dry skin, cutaneous pruritus, contusion and paronychia, as well as peripheral sensory neuropathy (11.1%). The any-grade events of skin disorders and peripheral sensory neuropathy were mostly observed in all patients. These events were controllable by preventive skin care and by withdrawal and dose reduction, respectively. Death due to adverse events was not observed. Adverse events did not require the withdrawal of this regimen. Conclusion: Based on the 18 patients studied, combined therapy with SOX+cetuximab was free of serious adverse events and could be safely administered by reducing the dose or temporarily suspending treatment, as required. These regimens seem to be promising for conversion therapy (4 out of 18 patients) because of good outcomes and an early response.

Thursday, April 28, 2016 9:59 AM|RADIĆ, J., KRUŠLIN, B., ŠAMIJA, M., ULAMEC, M., MILOŠEVIĆ, M., JAZVIĆ, M., ŠAMIJA, I., GRAH, J. J., BOLANČA, A., KUSIĆ, Z.|Anticancer Research recent issues|Labels: CRC

Background/Aim: Protein connexin 43 (Cx43), a part of intercellular gap junctions, is frequently down-regulated in tumors. The aim of the study was to compare Cx43 expression in primary colorectal tumors of patients with stage III and IV disease. Patients and Methods: Immunohistochemical expression of Cx43 was analyzed in 50 colorectal adenocarcinomas from surgically-treated patients of stage pT3N1-2 without metastases (M0) and 50 specimens of the same pTN stage from patients with synchronous liver metastases (M1). Association of Cx43 expression with clinicopathological factors and tumor site was also analyzed. Results: There was no significant difference in Cx43 expression between M0 and M1 tumor specimens (p=0.817), as well as in Cx43 expression between colonic and rectal tumors (p=0.116), respectively. Stromal expression of Cx43 was higher in M1 than in M0 tumors (p=0.004). Conclusion: Stromal Cx43 expression is possible indicator of metastatic potential of colorectal adenocarcinoma.

Thursday, April 28, 2016 9:59 AM|MITSELOU, A., GALANI, V., SKOUFI, U., ARVANITIS, D. L., LAMPRI, E., IOACHIM, E.|Anticancer Research recent issues|Labels: CRC

The Syndecan-1 protein plays a crucial role in cell proliferation, cell adhesion, cell migration and angiogenesis and, at the same time, its co-expression with E-cadherin is regulated during epithelial-mesenchymal transition (EMT). In colorectal cancer (CRC), the expression of syndecan-1, E-cadherin/β-catenin complex is frequently disturbed. Angiogenesis is critical for the growth and metastatic spread of tumors. In the present study, we focused on the expression of these biological molecules and their prognostic significance in human CRC. Formalin-fixed paraffin-embedded surgical specimens from 69 patients with CRC were immunostained for syndecan-1, E-cadherin, β-catenin, endoglin (CD105) and CD31 (platelet cell adhesion molecule (PCAM-1)). A significant association was found between syndecan-1 with E-cadherin (p<0.0001), as well with β-catenin (p<0.0001). High β-catenin expression appeared to reduce the risk of poor outcome. Endoglin microvascular density (MVD) count was correlated significantly with Dukes' stage (p<0.0001), vessel invasion (p<0.0001), lymph node metastasis (p=0.039), liver metastasis (p<0.0001), recurrence of disease (p=0.010) and poor survival rate (p<0.0001). Endoglin tumor epithelial cell expression was associated with E-cadherin, β-catenin and syndecan-1 (p=0.001, p=0.068 and p=0.005, respectively). In conclusion, changes in the pattern of expression of syndecan-1, EMT markers, E-cadherin/β-catenin, in association with endoglin (CD105), may be involved in tumor progression and prognosis of CRC patients. Further studies are needed to clarify the interaction between these proteins and tumor initiation and progression.

Thursday, April 28, 2016 9:59 AM|SADAHIRO, S., SUZUKI, T., TANAKA, A., OKADA, K., SAITO, G., KAMIJO, A., NAGASE, H.|Anticancer Research recent issues|Labels: HIF, CRC

Aim: The aim of the study was to identify biomarkers capable of predicting response to preoperative chemoradiotherapy (CRT) including S-1 or UFT for rectal cancer using biopsy specimens obtained before CRT (Pre-samples) and 7 days after the start of CRT (Day-7 samples). Materials and Methods: Preoperative CRT including S-1 or UFT was performed in 82 patients with locally advanced rectal cancer. The expression levels of 18 genes related to 5-fluorouracil, folate, and radiation in the Pre-samples and the Day-7 samples were evaluated using reverse transcription polymerase chain reaction (RT-PCR) assay. Results: The gene expression levels of hypoxia inducible factor 1 alpha subunit (HIF1A), dihydropyrimidine dehydrogenase (DPYD) and thymidine phosphorylase (TYMP) were found significantly increased in Day-7 samples compared to Pre-samples in responders, but not in non-responders. Conclusion: Increases in gene expression levels of TYMP, DPYD, and HIF1A in tumor tissues at 7 days after the start of CRT may be useful for predicting the efficacy of CRT including S-1 or UFT.

Saturday, March 19, 2016 10:51 AM|Oncotarget|MedWorm: Cancer Therapy|Comments|Labels: IGFR, CRC
Authors: Dowling CM, Phelan J, Callender JA, Cathcart MC, Mehigan B, McCormick P, Dalton T, Coffey JC, Newton AC, O'Sullivan J, Kiely PA Abstract Despite extensive efforts, cancer therapies directed at the Protein Kinase C (PKC) family of serine/threonine kinases have failed in clinical trials. These therapies have been directed at inhibiting PKC and have, in some cases, worsened disease outcome. Here we examine colon cancer patients and show not only that PKC Beta II is a tumour suppressor, but patients with low levels of this isozyme have significantly decreased disease free survival. Specifically, analysis of gene expression levels of all PKC genes in matched normal and cancer tissue samples from colon cancer patients revealed a striking down-regulation of the gene coding PKC Be...
Authors: Sun S, Cheng S, Zhu Y, Zhang P, Liu N, Xu T, Sun C, Lv Y Abstract Oncogene and non-oncogene addictions describe the phenomenon that tumor cells become reliant on certain genes for maintenance of malignancy. Reversal of these mutations profoundly affects tumor growth and survival, providing a fundamental rationale for development of targeted cancer therapy. However, inadequate knowledge on cancer signaling networks and lack of potential drug targets limited its clinical application. A screen was conducted using a custom small interfering RNA (siRNA) library in colorectal cancer (CRC). Transient knockdown followed by cell proliferation assays were performed to validate the essentiality of PRKDC (Protein Kinase, DNA-Activated, Catalytic Polypeptide) in CRC. Western blot analy...
Friday, March 11, 2016 4:00 PM|International Journal of Cancer|MedWorm: Cancer Therapy|Comments|Labels: CRC
Adjuvant chemotherapy can be considered in high‐risk stage II colon cancer comprising pT4, poor/undifferentiated grade, vascular invasion, emergency surgery and/or <10 evaluated lymph nodes (LNs). Adjuvant chemotherapy administration and its effect on survival was evaluated for each known risk factor. All patients with high‐risk stage II colon cancer who underwent resection and were diagnosed in the Netherlands between 2008 and 2012 were included. After stratification by risk factor(s) (vascular invasion could not be included), Cox regression was used to discriminate the independent association of adjuvant chemotherapy with the probability of death. Relative survival was used to estimate disease‐specific survival. A total of 4,940 of 10,935 patients with stage II colon cancer were...

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Sunday, March 6, 2016 4:00 PM|World Journal of Gastroenterology : WJG|MedWorm: Cancer Therapy|Comments|Labels: CRC
CONCLUSION: miR-145 is important in inhibiting colon cancer cell proliferation and migration. This is a good foundation for development of colon cancer therapy by targeting tumor suppressor miR-145. PMID: 26973415 [PubMed - in process] (Source: World Journal of Gastroenterology : WJG)

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Wednesday, March 2, 2016 4:00 PM|Life Sciences|MedWorm: Cancer Therapy|Comments|Labels: CRC, clinical trial
Publication date: 1 April 2016 Source:Life Sciences, Volume 150 Author(s): Martin Kello, David Drutovic, Martina Bago Pilatova, Vierka Tischlerova, Pal Perjesi, Jan Mojzis Aims Chalcones, naturally occurring open-chain polyphenols abundant in plants, have demonstrated antiproliferative activity in several cancer cell lines. In the present study, the potential anticancer activity of two synthetic analogues named Ch1 and Ch2 in colon cancer cell line was investigated. Main methods Antiproliferative activities of both synthetic analogues were assessed by Growth Inhibition Assay (MTT) and xCELLigence cell analysis. Apoptosis was assessed by annexin V/PI staining (early stage) or by DNA fragmentation (final stage). To study the cell death mechanism induced by tested substances, we asse...
Monday, February 22, 2016 4:00 PM|Bioorganic and Medicinal Chemistry Letters|Bioorganic and Medicinal Chemistry Letters via|Comments|Labels: WNT, CRC
Authors: Radwan AA, Al-Mohanna F, Alanazi FK, Manogaran PS, Al-Dhfyan A Abstract Cell surface molecule CD44 plays a major role in regulation of cancer stem cells CSCs on both phenotypic and functional level, however chemical inhibition approach of CD44 to targets CSCs is poorly studied. Herein, we report the discovery of certain N'-(2-oxoindolin-3-ylidene)-2-(benzyloxy)benzohydrazides as a novel inhibitor of CD44. Molecular docking study showed interference of the scaffold of these compounds with β-catenin/TCF-4 complex, building a direct relationship between CD44 inhibition and observed well-fitted binding domain. Compound 11a, most potent member elicits inhibition effect on TCF/LEF reporter activity conformed the involvement of Wnt pathway inhibition as a mechanism of action. Fu...
Friday, February 19, 2016 5:16 AM|Oncotarget|MedWorm: Cancer Therapy|Comments|Labels: STAT, CRC, liver cancer
Authors: Chongqiang Z, Wenlong W, Wenying Y, David J, Yina W, Haiyan M, Hui X, Hua Q, Z, Jiagao L, Sheng L, Chenglong L, Jiayuh L, Li L Abstract Signal Transducer and Activator of Transcription 3 (STAT3) is persistently activated in human liver and colon cancer cells and is required for cancer cell viability, survival and migration. Therefore, inhibition of STAT3 signaling may be a viable therapeutic approach for these two cancers. We recently designed a non-peptide small molecule STAT3 inhibitor, LY5, using in silico site-directed Fragment-based drug design (FBDD). The inhibitory effect on STAT3 phosphorylation, cell viability, migration and colony forming ability by LY5 were examined in human liver and colon cancer cells. We demonstrated that LY5 inhibited constitutive Inter...
Tuesday, February 16, 2016 4:00 PM|Journal of International Medical Research|MedWorm: Cancer Therapy|Comments|Labels: CRC
Conclusions The findings of this study suggest that SMC1A plays an oncogenic role in colorectal cancer and that it might be a promising target for colorectal cancer therapy. (Source: Journal of International Medical Research)
Monday, February 15, 2016 4:00 PM|BMB Reports|MedWorm: Cancer Therapy|Comments|Labels: EGFR, CRC, clinical trial
Authors: Joo D, Woo JS, Cho KH, Han SH, Min TS, Yang DC, Yun CH Abstract Type-specific features of cancer cells may strongly influence the effectiveness of therapeutic interventions. Here, we show that the spontaneous reactivation of extracellular signal-regulated kinase (ERK), distinct from conventional ERK activation, represents a potent mechanism for cancer cell survival. We studied ERK1/2 activation in vitro in SW480 colorectal cancer cells. Although ERK signaling tends to be only transiently activated, we observed the delayed reactivation of ERK1/2 in EGF-stimulated SW480 cells. This effect was observed even after EGF withdrawal. While phosphorylated ERK1/2 translocated into the nucleus following its primary activation, it remained in the cytoplasm during late-phase activation...
This meta-analysis evaluates the clinical evidence for the addition of traditional medicines (TMs) to oxaliplatin-based regimens for colorectal cancer (CRC) in terms of tumor response rate (TRR). Eight electronic databases were searched for randomized controlled trials of oxaliplatin-based chemotherapy combined with TMs compared to the same oxaliplatin-based regimen. Data on TRR from 42 randomized controlled trials were analyzed using Review Manager 5.1. Studies were conducted in China or Japan. Publication bias was not evident. The meta-analyses suggest that the combination of the TMs with oxaliplatin-based regimens increased TRR in the palliative treatment of CRC (risk ratio [RR] 1.31 [1.20-1.42], I2 = 0%). Benefits were evident for both injection products (RR 1.36 [1.18-1.57], I2 = 0%) ...
Monday, February 1, 2016 4:00 PM|FEBS Letters|MedWorm: Cancer Therapy|Comments|Labels: CRC
Cytochrome P450 2W1 (CYP2W1) is a colon tumor‐specific enzyme, suggested as a potential target for cancer therapy. In contrast to other endoplasmic reticulum P450s, we found completely inverted ER membrane topology of CYP2W1 using different approaches (redox sensitive luciferase assay and protease protection assay) and demonstrated that canonical CYP reductants, cytochrome P450 reductase, and cytochrome b5 cannot serve as electron donors for CYP2W1. Moreover, the reduced catalytic activity of the Asn177 mutant that is modified by glycan moieties in the wild‐type enzyme indicates a functional relevance of CYP2W1 glycosylation. (Source: FEBS Letters)
Clinical outcomes of colorectal cancer are influenced not by tumor size, but by spread into the bowel wall. Although assessment of serosal involvement is an important pathological feature for classification of colon cancer, its diagnostic consistency has been questioned. Using elastic staining, we assessed elastic laminal invasion (ELI) for more objective stratification of deep tumor invasion around the peritoneal surface. In addition, pathological characteristic features of marked tumor budding, fibrosis, and macrophage infiltration in the tumor area with ELI was elucidated. This characteristic tumor area was termed cancer microenvironment formed by peritoneal elastic laminal invasion (CMPI). We elucidated histoanatomical layer‐dependent heterogeneity of fibroblast in colonic tissue. Fu...
Although new classifications for neuroendocrine tumors were established by the World Health Organization, the current procedures and terms used in pathology laboratories are not known. A Web‐based questionnaire was distributed to 491 institutions affiliated with the Japanese Society for Cancer of the Colon and Rectum, and 150 participated. The questionnaires included questions regarding routine pathological reporting, staining, and assessment of neuroendocrine tumors. Next, the time taken to assess Ki‐67 index and mitotic count according to recommendation was evaluated to determine its feasibility. Most laboratories recorded diagnostic term, depth of invasion, size, lymph‐vascular invasion, Ki‐67 index, and mitotic count. However, only 32.2% reported tumor stage. Chromogranin A and...
Friday, January 22, 2016 2:15 PM|News-Medical.Net Cetuximab News Feed|Comments|Labels: BRAF, RAS, CRC
A post hoc analysis of the PETACC-8 trial has revealed an interaction between microsatellite instability and BRAF and KRAS mutation status when determining the prognosis of patients with resected stage III colon adenocarcinoma.
Sunday, January 10, 2016 4:00 PM|Pathology International|Pathology International via|Comments|Labels: CRC
This study examines the interobserver variability according to pathologists' experience and evaluates the influence of cytokeratin (CK) immunostaining in the assessment of budding. Hematoxylin‐eosin (HE) and CK‐immunostained slides of 40 cases with T1 primary colorectal cancer were examined. Budding grades were individually evaluated by 12 pathologists who we categorized into three groups by their experience (expert, with >10 years of experience (n = 4), senior, with 5–10 years (n = 4), and junior, < 5 years (n = 4)). The results revealed a tendency for the more experienced pathologists to assign higher budding grades compared to the less‐experienced pathologists. In the junior group, the interobserver variability obtained with HE slides was poor, but it was markedly improved...
Wednesday, January 6, 2016 4:00 PM|Molecular Cancer Therapeutics|MedWorm: Cancer Therapy|Comments|Labels: P53, CRC
TP53, a well-known tumour suppressor gene, is frequently inactivated by mutation or deletion in a majority of human tumours. A tremendous effort has been made to restore p53 activity in cancer therapies. However, no effective p53-based therapy has been successfully translated into clinical cancer treatment due to the complexity of p53 regulators and their poor drugability. Here, we demonstrate that genomic deletion of TP53 frequently encompasses neighbouring essential genes, rendering cancer cells with hemizygous TP53 deletion vulnerable to further suppression of such genes. POLR2A is identified as such an essential house-keeping gene that is virtually co-deleted with TP53 in many types of human cancer. It encodes the largest and catalytic subunit of RNA polymerase II complex, which is spe...