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Oncology Intelligence

BRAF
BRAF BRAF(3)

The RAF family of serine/threonine-specific protein kinases is involved in directing cell growth.(1, 2) BRAF plays a role in regulating the MAP kinase/ERKs signaling pathway, which affects cell division, differentiation, and secretion.(2) Signaling through this pathway is driven by RAS family GTPases, including HRAS, KRAS, and NRAS. RAF kinases phosphorylate and activate MEK1 and MEK2, which subsequently phosphorylate and activate ERK1 and ERK2.(3)
RAS and BRAF mutations lead to the constitutive activation of EGFR signaling.(4) Acquired mutations in BRAF gene have also been found in cancers, including NHL, CRC, melanoma, thyroid, ovarian, NSCLC, and lung.(2, 5)
New targeted agents directed against the RAS-RAF-MEK-ERK pathway show promising activity in early clinical development and particular interest is focused on selective inhibitors of mutant BRAF.(3-5)

Drugs/Indications
Marketed Drugs
Generic Code Old Code Brand Company Indication trials
vemurafenib RG7204 RO5185426, PLX4032 Zelboraf Roche Mkt: melanoma; P3: thyroid: P2: leukemia, solid; P1: brain trials
dabrafenib GSK2118436 Tafinlar GSK Mkt: melanoma; P2: NSCLC, thyroid, solid, brain mets; P1/2: CRC, ovarian, epithelial trials
Trial Drugs/Indications
Generic Code Old Code Brand Company Indication trials
encorafenib LGX818 Array, Novartis P3: melanoma; P2: solid, hem, NSCLC; P1/2: CRC trials
RAF-265 CHIR-265 Novartis P2: melanoma; P1: solid trials
BMS-908662 XL281 BMS P1/2: CRC; P1: melanoma trials
PLX8394 Plexxikon P1/2: melanoma, thyroid, CRC, NSCLC, cholangiocarcinoma, histiocytosis, hairy cell leukemia trials
ARQ 736 ArQule P1: solid trials
LY3009120 Eli Lilly P1: melanoma, NSCLC, CRC, solid trials
MLN2480 Millennium P1: solid, melanoma trials
RO5212054 PLX3603 Roche P1: solid, CRC, melanoma trials
Failed Drugs/Indications
Generic Code Old Code Brand Company Indication trials
LErafAON NeoPharm Last trial started in 2004; P1: solid trials


News
References

1. Qiao J KT, Willmon C, Galivo F, Wongthida P, Diaz RM, Thompson J, Ryno P, Barber GN, Chester J, Selby P, Harrington K, Melcher A, Vile RG. Purging metastases in lymphoid organs using a combination of antigen-nonspecific adoptive T cell therapy, oncolytic virotherapy and immunotherapy. Nat Med. 2008;14:37-44. PMCID: 18066076.

2. Davies H BG, Cox C, Stephens P, Edkins S, Clegg S, Teague J, Woffendin H, Garnett MJ, Bottomley W, Davis N, Dicks E, Ewing R, Floyd Y, Gray K, Hall S, Hawes R, Hughes J, Kosmidou V, Menzies A, Mould C, Parker A, Stevens C, Watt S, Hooper S, Wilson R, Jayatilake H, Gusterson BA, Cooper C, Shipley J, Hargrave D, Pritchard-Jones K, Maitland N, Chenevix-Trench G, Riggins GJ, Bigner DD, Palmieri G, Cossu A, Flanagan A, Nicholson A, Ho JW, Leung SY, Yuen ST, Weber BL, Seigler HF, Darrow TL, Paterson H, Marais R, Marshall CJ, Wooster R, Stratton MR, Futreal PA. Mutations of the BRAF gene in human cancer. Nature. 2002;417(6892):949-54. PMCID: 12068308.

3. Corcoran RB SJ, Engelman JA. Potential therapeutic strategies to overcome acquired resistance to BRAF or MEK inhibitors in BRAF mutant cancers. Oncotarget. 2011;4:336-46. PMCID: 21505228.

4. T. Y. Are KRAS/BRAF mutations potent prognostic and/or predictive biomarkers in colorectal cancers? Anticancer Agents Med Chem. 2012;12(2):163-71. PMCID: 22043994.

5. Arkenau HT KR, Long GV. Targeting BRAF for patients with melanoma. Br J Cancer. 2011;104(3):392-8. PMCID: 21139585.

6. Nick J. Dibb SMDCDM. Switching on kinases: oncogenic activation of BRAF and the PDGFR family. Nature Reviews Cancer. 2004;4:718-27.



News
Wednesday, October 26, 2016 3:00 PM|Onclive Articles|Labels: BRAF
Single-agent olaparib significantly improved progression-free survival compared with placebo in the maintenance setting for patients with advanced BRAF-positive ovarian cancer.
Tuesday, October 25, 2016 4:31 PM|PR Newswire Association LLC.|PR Newswire: Pharmaceuticals|Attachments|Labels: BRAF, melanoma, clinical trial

Array BioPharma. (PRNewsFoto/Array BioPharma Inc.) (PRNewsFoto/)BOULDER, Colo., and CASTRES, France, Oct. 25, 2016 /PRNewswire/ -- Array BioPharma (NASDAQ: ARRY) and Pierre Fabre announced today that results from the Phase 3 COLUMBUS trial of binimetinib and encorafenib in BRAF-mutant melanoma will be presented at the 2016 Society for Melanoma...


Tuesday, October 25, 2016 10:20 AM|Pietrantonio, F., Vernieri, C., Siravegna, G., Mennitto, A., Berenato, R., Perrone, F., Gloghini, A., Tamborini, E., LONARDI, S., Morano, F., Picciani, B., Busico, A., Volpi, C. C., Martinetti, A., Battaglin, F., Bossi, I., Pellegrinelli, A., Milione, M., Cremolini, C., Di Bartolomeo, M., Bardelli, A., de Braud, F.|Clinical Cancer Research Online First Articles|Labels: BRAF, EGFR, CRC

Purpose Even if RAS-BRAF wild-type and HER2/MET negative mCRC patients frequently respond to anti-EGFR monoclonal antibodies, acquired resistance almost invariably occurs. Mechanisms of resistance to EGFR blockade include the emergence of KRAS, NRAS and EGFR extracellular domain mutations as well as HER2/MET alterations. However, these findings derive from retrospective studies that analyzed one single resistance mechanism at a time; moreover, it is still unclear how molecular heterogeneity affects clonal evolution in patients. In this work, we aimed at extensively characterizing and correlating the molecular characteristics of tissue- and blood-based data in a prospective cohort of mCRC patients who received anti-EGFR antibodies. Experimental design Twenty-two RAS-BRAF wild-type, HER2/MET negative mCRC patients progressing on anti-EGFR therapy after initial response underwent re-biopsy. Next-generation sequencing and SISH/IHC analyses were performed both on archival tumors and post-progression samples. ctDNA molecular profiles were obtained in matched tissue-plasma samples. Results RAS mutations and HER2/MET amplification were the most frequently detected resistance mechanisms in both tissue and blood sample analysis. On the other hand, BRAF and EGFR ectodomain mutations were much rarer. Patients with acquired MET amplification showed worse PFS on anti-EGFRs. We detected both intra-lesion heterogeneity, as suggested by co-occurrence of different resistance mechanisms in the same sample, and inter-lesion heterogeneity. The combined analysis of tissue and blood (ctDNA) results highlights the complexity of clonal evolution triggered by EGFR blockade. Conclusions Our results indicate that it may be extremely challenging to target the complex landscape of molecular heterogeneity associated with emergence of resistance to targeted therapies in mCRC.

Friday, October 14, 2016 9:31 AM|Qian Shi; Jeffrey P Meyers; Timothy S Maughan; Richard A Adams; Matthew T Seymour; Leonard Saltz; Cornelis J A Punt; Miriam Koopman; Christophe Tournigand; Niall C Tebbutt; Eduardo Diaz-Rubio; John Souglakos; Alfredo Falcone; Benoist Chibaudel; Volker Heinemann; Joseph Moen; Aimery De Gramont; Daniel J Sargent; Axel Grothey|JournalTOCs API - The Lancet Oncology (50 articles)|Labels: BRAF, CRC

Prognosis of patients with peritoneal metastatic colorectal cancer given systemic therapy: an analysis of individual patient data from prospective randomised trials from the Analysis and Research in Cancers of the Digestive System (ARCAD) database
Jan Franko Qian Shi; Jeffrey P Meyers; Timothy S Maughan; Richard A Adams; Matthew T Seymour; Leonard Saltz; Cornelis J A Punt; Miriam Koopman; Christophe Tournigand; Niall C Tebbutt; Eduardo Diaz-Rubio; John Souglakos; Alfredo Falcone; Benoist Chibaudel; Volker Heinemann; Joseph Moen; Aimery De Gramont; Daniel J Sargent; Axel Grothey
The Lancet Oncology, Vol. , No. (2016) pp. -
Publication date: Available online 12 October 2016 Source:The Lancet Oncology Author(s): Jan Franko, Qian Shi, Jeffrey P Meyers, Timothy S Maughan, Richard A Adams, Matthew T Seymour, Leonard Saltz, Cornelis J A Punt, Miriam Koopman, Christophe Tournigand, Niall C Tebbutt, Eduardo Diaz-Rubio, John Souglakos, Alfredo Falcone, Benoist Chibaudel, Volker Heinemann, Joseph Moen, Aimery De Gramont, Daniel J Sargent, Axel Grothey Background Patients with peritoneal metastatic colorectal cancer have reduced overall survival compared with patients with metastatic colorectal cancer without peritoneal involvement. Here we further investigated the effect of the number and location of metastases in patients receiving first-line systemic chemotherapy. Methods We analysed individual patient data for previously untreated patients enrolled in 14 phase 3 randomised trials done between 1997 and 2008. Trials were included if protocols explicitly pre-specified and solicited for patients with peritoneal involvement in the trial data collection process or had done a formal peritoneum-focused review of individual pre-treatment scans. We used stratified multivariable Cox models to assess the prognostic associations of peritoneal metastatic colorectal cancer with overall survival and progression-free survival, adjusting for other key clinical-pathological factors (age, sex, Eastern Cooperative Oncology Group (ECOG) performance score, primary tumour location [colon vs rectum], previous treatment, and baseline BMI). The primary endpoint was difference in overall survival between populations with and without peritoneal metastases. Findings Individual patient data were available for 10 553 patients. 9178 (87%) of 10 553 patients had non-peritoneal metastatic colorectal cancer (4385 with one site of metastasis, 4793 with two or more sites of metastasis), 194 (2%) patients had isolated peritoneal metastatic colorectal cancer, and 1181 (11%) had peritoneal metastatic colorectal cancer and other organ involvement. These groups were similar in age, ethnic origin, and use of targeted treatment. Patients with peritoneal metastatic colorectal cancer were more likely than those with non-peritoneal metastatic colorectal cancer to be women (565 [41%] of 1371 vs 3312 [36%] of 9169 patients; p=0·0003), have colon primary tumours (1116 [84%] of 1334 patients vs 5603 [66%]; p<0·0001), and have performance status of 2 (136 [10%] vs 521 [6%]; p<0·0001). We recorded a higher proportion of patients with mutated BRAF in patients with peritoneal-only (eight [18%] of 44 patients with available data) and peritoneal metastatic colorectal cancer with other sites of metastasis (34 [12%] of 289), compared with patients with non-peritoneal metastatic colorectal cancer (194 [9%] of 2230; p=0·028 comparing the three groups). Overall survival (adjusted HR 0·75, 95% CI 0·63–0·91; p=0·003) was better in patients with isolated non-peritoneal sites than in those with isolated peritoneal metastatic colorectal cancer. Overall survival of patients with two of more non-peritoneal sites of metastasis (adjusted HR 1·04, 95% CI 0·86–1·25, p=0.69) and those with peritoneal metastatic colorectal cancer plus one other site of metastasis (adjusted HR 1·10, 95% CI 0·89–1·37, p=0·37) was similar to those with isolated peritoneal metastases. Compared with patients with isolated peritoneal metastases, those with peritoneal metastases and two or more additional sites of metastasis had the shortest survival (adjusted HR 1·40; CI 1·14–1·71; p=0·0011). Interpretation Patients with peritoneal metastatic colorectal cancer have significantly shorter overall survival than those with other isolated sites of metastases. In patients with several sites of metastasis, poor survival is a function of both increased number of metastatic sites and peritoneal involvement. The pattern of metastasis and in particular, peritoneal involvement, results in prognostic heterogeneity of metastatic colorectal cancer. Funding None.

Friday, October 14, 2016 12:05 AM|Ryan, M. B., Finn, A. J., Pedone, K. H., Thomas, N. E., Der, C. J., Cox, A. D.|Molecular Cancer Research recent issues|Labels: BRAF, MapK, melanoma, skin cancer

Recently, we identified that PREX1 overexpression is critical for metastatic but not tumorigenic growth in a mouse model of NRAS-driven melanoma. In addition, a PREX1 gene signature correlated with and was dependent on ERK MAPK activation in human melanoma cell lines. In the current study, the underlying mechanism of PREX1 overexpression in human melanoma was assessed. PREX1 protein levels were increased in melanoma tumor tissues and cell lines compared with benign nevi and normal melanocytes, respectively. Suppression of PREX1 by siRNA impaired invasion but not proliferation in vitro. PREX1-dependent invasion was attributable to PREX1-mediated activation of the small GTPase RAC1 but not the related small GTPase CDC42. Pharmacologic inhibition of ERK signaling reduced PREX1 gene transcription and additionally regulated PREX1 protein stability. This ERK-dependent upregulation of PREX1 in melanoma, due to both increased gene transcription and protein stability, contrasts with the mechanisms identified in breast and prostate cancers, in which PREX1 overexpression was driven by gene amplification and HDAC-mediated gene transcription, respectively. Thus, although PREX1 expression is aberrantly upregulated and regulates RAC1 activity and invasion in these three different tumor types, the mechanisms of its upregulation are distinct and context dependent.

Implications: This study identifies an ERK-dependent mechanism that drives PREX1 upregulation and subsequent RAC1-dependent invasion in BRAF- and NRAS-mutant melanoma. Mol Cancer Res; 14(10); 1009–18. ©2016 AACR.

Thursday, October 13, 2016 12:00 PM|E.|JournalTOCs API - Blood (22 articles)|Labels: BRAF, leukemia

BRAF V600E mutation in hairy cell leukemia: from bench to bedside
Falini, B Martelli, M. P, Tiacci, E.
Blood, Vol. 128, No. 15 (2016) pp. 1918 - 1927
Hairy cell leukemia (HCL) is a distinct clinicopathological entity whose underlying genetic lesion has remained a mystery for over half a century. The BRAF V600E mutation is now recognized as the causal genetic event of HCL because it is somatic, present in the entire tumor clone, detectable in almost all cases at diagnosis (encompassing the whole disease spectrum), and stable at relapse. BRAF V600E leads to the constitutive activation of the RAF-MEK-extracellular signal-regulated kinase (ERK) signaling pathway which represents the key event in the molecular pathogenesis of HCL. KLF2 and CDNK1B (p27) mutations may cooperate with BRAF V600E in promoting leukemic transformation. Sensitive molecular assays for detecting BRAF V600E allow HCL (highly responsive to purine analogs) to be better distinguished from HCL-like disorders, which are treated differently. In vitro preclinical studies on purified HCL cells proved that BRAF and MEK inhibitors can induce marked dephosphorylation of MEK/ERK, silencing of RAF-MEK-ERK pathway transcriptional output, loss of the HCL-specific gene expression profile signature, change of morphology from "hairy" to "smooth," and eventually apoptosis. The overall response rate of refractory/relapsed HCL patients to the BRAF inhibitor vemurafenib approached 100%, with 35% to 40% complete remissions (CRs). The median relapse free-survival was about 19 months in patients who had achieved CR and 6 months in those who had obtained a partial response. Future therapeutic perspectives include: (1) combining BRAF inhibitors with MEK inhibitors or immunotherapy (anti-CD20 monoclonal antibody) to increase the percentage of CRs and (2) better understanding of the molecular mechanisms underlying resistance of HCL cells to BRAF inhibitors.

Wednesday, October 12, 2016 3:26 PM|E. Klimi|JournalTOCs API - Journal of Investigative Dermatology (875 articles)|Labels: BRAF, melanoma, skin cancer

458 Leopard syndrome associated with metastatic malignant melanoma of unknown origin
E. Klimi
Journal of Investigative Dermatology, Vol. 136, No. 9 (2016) pp. S238 -
Leopard syndrome is a rare autosomal dominant condition associating lentigines, electrocardiographic conduction abnormalities, ocular hypertelorism, pulmonary stenosis, abnormal genitalia, retardation of growth and sensorineural deafness. 90% of Leopard syndrome cases -type1-are associated with mutations of PTPN11 gene while the rest 10% are associated either with mutations in BRAF or RAF gene. A 63 year old woman had a cerebral hematoma evacuated surgically and the histology found HBM positive cells in the debris of the liguid Skin examination showed lentigines on exposed and non exposed body areas, facial dysmorphia, normal ECG, normal X-Rays, normal liver echography and ORL and gynaecological examination also normal.

Wednesday, October 12, 2016 3:26 PM|G. Schuler|JournalTOCs API - Journal of Investigative Dermatology (875 articles)|Labels: BRAF, melanoma

510 SPPL3-mediated ADAM10 Activation is a Key Step in the evolution of BRAFV600E+ Superficial Spreading Melanomas (SSM)
A.S. Baur C. Ostalecki, J. Lee, G. Schuler
Journal of Investigative Dermatology, Vol. 136, No. 9 (2016) pp. S247 -
Malignant melanocyte transformation is characterized by the appearance of specific mutations, notably in the BRAF and PI3-kinase pathway. However, little is known how these mutations cooperate on the protein level. Using the multi-epitope-ligand-cartography (MELC)-technology, we analyzed protein expression profiles (PEP) in different nevi and BRAFV600E+ superficial spreading melanomas (SSM) for key transformation events. Surprisingly, these PEPs differed only in expression levels of specific antigens.

Wednesday, October 12, 2016 3:26 PM|T. Ruksha|JournalTOCs API - Journal of Investigative Dermatology (875 articles)|Labels: BRAF, melanoma, skin cancer

507 Clinicopathological features and NRAS gene mutation status in patients with primary cutaneous melanoma
M. Aksenenko T. Ruksha
Journal of Investigative Dermatology, Vol. 136, No. 9 (2016) pp. S247 -
NRAS oncogene mutations are the second most common mutation type in cutaneous melanoma. Large majority of these mutations affect NRAS exon 3, codons 59, 60,61 or 62. NRAS mutations are often present in melanomas arising from sites of chronic sun damage. The aim of this study was to evaluate NRAS mutations exon 3 frequency in BRAF-negative melanomas. Genomic DNA was extracted from biopsy specimens with suitable percentage of tumor cells. BRAF V600 mutations were estimated by real-time PCR-based allele-specific DNA test.

Wednesday, October 12, 2016 3:26 PM|M. Fargnoli|JournalTOCs API - Journal of Investigative Dermatology (875 articles)|Labels: BRAF, telomerase, melanoma, skin cancer

494 Heterogeneous mutational status of melanomas in multiple primary melanoma patients
C. Pellegrini C. Martorelli, G. Cipolloni, L. Di Nardo, A. Antonini, M. Maturo, M. Fargnoli
Journal of Investigative Dermatology, Vol. 136, No. 9 (2016) pp. S244 -
Multiple primary melanomas (MPM) develop in about 5% of sporadic melanoma patients and in up to 19% of melanoma patients with a positive family history. Many genetic alterations, including predisposing and/or somatic mutations, may contribute to the development of MPM and data on the genetic diversity of MPMs are limited. We aimed to assess the frequency and distribution of BRAF, NRAS and TERT promoter somatic mutations in subsequent melanomas of the same patient and evaluate the association of somatic alterations with germline mutational profile of MPM patients.

Wednesday, October 12, 2016 3:26 PM|M. Fargnoli|JournalTOCs API - Journal of Investigative Dermatology (875 articles)|Labels: BRAF, melanoma, skin cancer

503 BRAF-V600 Mutation discordance in Primary and Metastatic Melanoma
C. Martorelli C. Pellegrini, L. Di Nardo, A. Marinucci, A. Antonini, M. Maturo, M. Fargnoli
Journal of Investigative Dermatology, Vol. 136, No. 9 (2016) pp. S246 -
BRAF V600 mutation is the most common mutation in cutaneous melanoma and is currently the target mutation of molecular therapy with selective BRAF inhibitors in metastatic melanoma patients. The diversity between primary and metastatic melanoma lesions in individual patients is known; however, the molecular and genetic differences remain unclear. Our aim was to investigate the mutational status of primary melanomas and of corresponding metastatic lesions evaluating the genetic discordance rate for BRAF mutations.

Wednesday, October 12, 2016 3:26 PM|Fred Hutchinson Cancer Research Center/UW Medicine Cancer Consortium - Clinical Trials|Labels: BRAF, brain cancer, clinical trial
This is a multicenter, safety and pharmacokinetic trial to determine the MTD and/or select a recommended phase 2 dose (RP2D) of vemurafenib in children with recurrent or refractory gliomas containing the BRAFV600E mutation.
Wednesday, October 12, 2016 3:26 PM|Fred Hutchinson Cancer Research Center/UW Medicine Cancer Consortium - Clinical Trials|Labels: BRAF, skin cancer, clinical trial
This is a 2-part, study to determine the safety, tolerability and pharmacokinetics of oral dabrafenib in children and adolescent subjects with advanced BRAF V600 mutation-positive solid tumors. Part 1 (dose escalation study) will identify the recommended Part 2 (tumor-specific expansion study) dose and regimen using a dose-escalation procedure. Approximately 6 to 18 subjects will participate in Part 1 and will receive a starting dose of 3 mg/kg and dose will deescalate or escalate between 1.5 milligram (mg)/kilogram (kg) and 6 mg/kg. Up to 6 subjects will be enrolled at one dose level dependent upon the number of subjects at the current dose level, the number of subjects who have experienced a dose limiting toxicity (DLT) at the current dose level, and the number of subjects enrolled but with data pending at the current dose level. Escalation may proceed until either a maximum tolerated dose (MTD) is established, or until the dose in which the median pharmacokinetic parameters consistent with exposure in adults are achieved. Cohorts may be added in order to evaluate additional dose levels. Part 2 consists of four disease-specific cohorts of subjects with tumors known to have BRAF V600 activation (pediatric low-grade gliomas, pediatric high-grade gliomas, Langerhans cell histiocytosis [LCH], and other tumors such as melanoma and papillary thyroid carcinoma [PTC]). Each cohort will enroll at least 10 subjects with a pre-dose and at least 1 post-dose disease assessment. In both the parts of the study, on Day 1, a single first dose will be administered, and repeat dosing will begin on Day 2. PK sampling will be performed on Day 1 and Day 15 for subjects >=25 kg in weight. For subjects <25 kg and >=10 kg in weight, blood samples for PK analysis will be collected on Day 1 and Day 15. For subjects <10kg in weight, blood samples for PK analysis will be collected after repeated administration on Day 15 only. Safety and tolerability will be assessed throughout the study. Treatment with dabrafenib will be continued until disease progression or until no clinical benefit or development of an unacceptable toxicity, or until they withdraw consent or begin a new therapy. At the end of treatment, a final study visit will occur.
Wednesday, October 12, 2016 3:26 PM|Seung Tae Kim, Su Jin Lee, Se Hoon Park, Joon Oh Park, Ho Yeong Lim, Won Ki Kang, Jeeyun Lee, Young Suk Park|Journal of Cancer|Labels: BRAF

Background: We have conducted molecular profiling through a high-throughput molecular test as part of our clinical practice for patients with advanced gastrointestinal (GI) cancer or rare cancers including gastroenteropancreatic neuroendocrine tumors (GEP-NETs). Herein, we report on the molecular characterization of 14 metastatic GEP-NET patients.

Methods: We conducted the Ion AmpliSeq Cancer Hotspot Panel v2 (detecting 2,855 oncogenic mutations in 50 commonly mutated genes) and nCounter Copy Number Variation Assay, which was designed with 21 genes based on available targeted agents, as a high throughput genomic platform in 14 patients with metastatic GEP-NETs.

Results: Among the 14 GEP-NET patients analyzed in this study, 8 patients had grade III neuroendocrine carcinoma (NEC) and 6 had grade I/II NET. Primary sites included pancreas (n=3), small intestine and ascending colon (n=3), distal colon and rectum (n=5), and unknown primary origin (n=3). The most common metastatic site was the liver. Of 14 GEP-NET patients available for mutational profiling, 7 (50.0%) patients had one or more aberrations detected. Common aberrations were as follows: SMARCB1 mutation (n=2), TP53 mutation (n=2), STK11 mutation (n=1), RET mutation (n=1), and BRAF mutation (n=1). Gene amplification by nCounter was detected in only 1 patient, showing CCNE1 amplification, and this patient also had a TP53 mutation.

Conclusions: This high throughput genomic test may be useful to identify new drug targets in metastatic GEP-NET patients. Currently, we plan to conduct further genomic analysis to develop predictive and prognostic biomarkers in a larger number of GEP-NET patients.

Wednesday, October 12, 2016 3:24 PM|Andrea L. George, Robert Suriano, Shilpi Rajoria, Maria C. Osso, Neha Tuli, Elyse Hanly, Jan Geliebter, Angelo N. Arnold, Marc Wallack, Raj K. Tiwari|Journal of Cancer (RSS 2.0)|Labels: BRAF, MapK, melanoma, skin cancer, regulatory

Over expression of various immunogenic melanoma associated antigens (MAAs) has been exploited in the development of immunotherapeutic melanoma vaccines. Expression of MAAs such as MART-1 and gp100 is modulated by the MAPK signaling pathway, which is often deregulated in melanoma. The protein BRAF, a member of the MAPK pathway, is mutated in over 60% of melanomas providing an opportunity for the identification and approval by the FDA of a small molecule MAPK signaling inhibitor PLX4032 that functions to inactivate mutant BRAFV600E.

To this end, we characterized five patient derived primary melanoma cell lines with respect to treatment with PLX4032. Cells were treated with 5μM PLX4032 and harvested. Western blotting analysis, RT-PCR and in vitro transwell migration and invasion assays were utilized to determine treatment effects. PLX4032 treatment modulated phosphorylation of signaling proteins belonging to the MAPK pathway including BRAF, MEK, and ERK and abrogated cell phenotypic characteristics such as migration and invasion. Most significantly, PLX4032 led to an up regulation of many MAA proteins in three of the four BRAF mutated cell lines, as determined at the protein and RNA level. Interestingly, MAGE-A1 protein and mRNA levels were reduced upon PLX4032 treatment in two of the primary lines.

Taken together, our findings suggest that the BRAFV600E inhibitor PLX4032 has therapeutic potential over and above its known target and in combination with specific melanoma targeting vaccine strategies may have further clinical utility.

Wednesday, October 12, 2016 3:24 PM|Linli Zhou, Kun Yang, Thomas Andl, R. Randall Wickett, Yuhang Zhang|Journal of Cancer (RSS 2.0)|Labels: BRAF, GIST, melanoma, skin cancer

Melanoma is known as an exceptionally aggressive and treatment-resistant human cancer. Although a great deal of progress has been made in the past decade, including the development of immunotherapy using immune checkpoint inhibitors and targeted therapy using BRAF, MEK or KIT inhibitors, treatment for unresectable stage III, stage IV, and recurrent melanoma is still challenging with limited response rate, severe side effects and poor prognosis, highlighting an urgent need for discovering and designing more effective approaches to conquer melanoma. Melanoma is not only driven by malignant melanocytes, but also by the altered communication between neoplastic cells and non-malignant cell populations, including fibroblasts, endothelial and inflammatory cells, in the tumor stroma. Infiltrated and surrounding fibroblasts, also known as cancer-associated fibroblasts (CAFs), exhibit both phenotypical and physiological differences compared to normal dermal fibroblasts. They acquire properties of myofibroblasts, remodel the extracellular matrix (ECM) and architecture of the diseased tissue and secrete chemical factors, which all together promote the transformation process by encouraging tumor growth, angiogenesis, inflammation and metastasis and contribute to drug resistance. A number of in vitro and in vivo experiments have shown that stromal fibroblasts promote melanoma cell proliferation and they have been targeted to suppress tumor growth effectively. Evidently, a combination therapy co-targeting tumor cells and stromal fibroblasts may provide promising strategies to improve therapeutic outcomes and overcome treatment resistance. A significant benefit of targeting CAFs is that the approach aims to create a tumor-resistant environment that inhibits growth of melanomas carrying different genetic mutations. However, the origin of CAFs and precise mechanisms by which CAFs contribute to melanoma progression and drug resistance remain poorly understood. In this review, we discuss the origin, activation and heterogeneity of CAFs in the melanoma tumor microenvironment and examine the contributions of stromal fibroblasts at different stages of melanoma development. We also highlight the recent progression in dissecting and characterizing how local fibroblasts become reprogrammed and build a dynamic yet optimal microenvironment for tumors to develop and metastasize. In addition, we review key developments in ongoing preclinical studies and clinical applications targeting CAFs and tumor-stroma interactions for melanoma treatment.

Wednesday, October 12, 2016 3:24 PM|Sofia Lampaki, George Lazaridis, Konstantinos Zarogoulidis, Ioannis Kioumis, Antonis Papaiwannou, Katerina Tsirgogianni, Anastasia Karavergou, Theodora Tsiouda, Vasilis Karavasilis, Lonny Yarmus, Kaid Darwiche, Lutz Freitag, Antonios Sakkas, Angeliki Kantzeli, Sofia Baka, Wolfgang Hohenforst-Schmidt, Paul Zarogoulidis|Journal of Cancer (RSS 2.0)|Labels: ALK, BRAF, EGFR, lung cancer, clinical trial

Historical, the non-small cell lung cancer (NSCLC) was as a united disease entity and the chemotherapy to the metastatic cancer had limited results. Recent studies for the metastatic non-small cell lung cancer led to the ascertainment that the NSCLC does not constitute exclusively a disease entity, but different neoplasms guided from different molecular paths, different biological behavior and at extension requires different confrontation. Thus the new direction for the therapeutic approach of NSCLC is henceforth the most individualized approach based on the activated molecular paths of tumor. Distinct subtypes of NSCLC are driven by a specific genetic alteration, like EGFR, ALK, ROS1 or BRAF mutations, and these genetic alterations are sensitized to the inhibition of specific oncogenic pathways. The benefit from the use of tyrosine kinase inhibitors in patients with EGFR mutations it was confirmed by six randomized studies of phase III that investigated the role of gefitinib, erlotinib and afatinib. In these studies the response rates vary in the impressive percentages from 55% to 86% and were connected with a remarkable median progression free survival of approximately 8 to 13 months, and with better quality of life compared to that of chemotherapy. In early stages NSCLC is needed the individualization of systemic treatment in order to reduce toxicity that is observed in the classic chemotherapy and to impact outcome. The role of EGFR TKI's has been evaluated in the adjuvant chemotherapy in early stage resected NSCLC. The data from these studies suggest that adjuvant TKI therapy might not increase the overall survival, but delay the recurrences. Prospective trials restricted to EGFR or ALK driven NSCLC subsets potentially offering the opportunity for a definitive answer in early disease adjuvant setting (ALCHEMIST) or as induction treatment before stage III chemo-radiotherapy (RTOG 1210/Alliance 31101), are ongoing. Ongoing prospective trials may offer the opportunity for a definitive answer of the role of tyrosine kinase inhibitors in induction treatment before chemo-radiotherapy or in early disease adjuvant therapy.

Wednesday, October 12, 2016 3:23 PM|admin|Advances in Melanoma Resource Centre - European Journal of Cancer|Labels: BRAF, MapK, melanoma, skin cancer
Abstract BACKGROUND The degree of response and the duration of survival of patients treated with mitogen-activated protein kinase (MAPK) inhibitors are highly variable. Whether baseline clinicopathologic factors can predict the clinical course with...
Wednesday, October 12, 2016 3:23 PM|Hyunsu Lee, Jae-Ho Lee, Dae-Kwang Kim, In-Jang Choi, Ilseon Hwang, Yu-Na Kang, Shin Kim|International Journal of Medical Sciences|Labels: AKT, BRAF

Colorectal cancer is a heterogeneous disorder than arises via multiple distinct pathways, from tubular adenomas (TAs) and sessile serrated adenomas (SSAs), which are clinically, morphologically, and molecularly different. We examined PIK3CA amplification in colorectal precancerous legions, including TAs and SSAs. DNA was isolated from paired normal and tumoral tissues in 64 TAs and 32 SSAs. PIK3CA amplification, KRAS mutation, and BRAF mutation were analyzed by real-time PCR and pyrosequencing. PIK3CA amplification was found in 25% of TAs and 9.4% of SSAs, respectively. KRAS and BRAF mutations were mutually exclusive in both TAs and SSAs. In TAs, PIK3CA amplification was associated with left side and it was mutually exclusive with KRAS mutation. These results suggest that PIK3CA amplification may be early and important event in colorectal carcinogenesis and may drive the development of left-side TAs independently with KRAS mutation.

Wednesday, October 12, 2016 12:57 PM|Xi, L., Pham, T. H.-T., Payabyab, E. C., Sherry, R. M., Rosenberg, S. A., Raffeld, M.|Clinical Cancer Research Online First Articles|Labels: BRAF, melanoma, skin cancer, biomarker diagnostic

Purpose: Adoptive transfer of activated autologous tumor-infiltrating lymphocytes (TIL) can mediate complete, durable regressions in patients with metastatic melanoma. Responding patients generally do not have significant changes in noncutaneous RECIST targets before 30 to 60 days following TIL infusion, and complete responses are often not confirmed for 1 to 2 years. There is a critical need for a biomarker that can provide early information regarding the likelihood and duration of a response to enable rational decisions about altering therapy. We wished to evaluate the role of circulating tumor DNA (ctDNA) in separating responding from nonresponding patients.

Experimental Design: We studied BRAF V600E ctDNA levels by a sensitive allele-specific PCR assay in 388 serum samples from 48 patients who received TIL immunotherapy at the NCI and correlated differences in the dynamic patterns of their ctDNA measurements with response outcomes.

Results: A strong correlation was found between the presence or absence of an early serum peak of V600E ctDNA, and the likelihood of an objective response. Furthermore, patients that developed an early ctDNA peak and cleared their serum of V600E ctDNA were highly likely to achieve a complete response over the next 1 to 2 years. Patients that showed no peak of V600E ctDNA failed to achieve an objective response, with one exception.

Conclusions: We show that the dynamic changes occurring in BRAF V600E ctDNA levels within the first month following T-cell transfer immunotherapy in metastatic melanoma can be used to rapidly identify responding from nonresponding patients, potentially allowing clinicians to make critical treatment-related decisions in a more timely manner. These data also suggest that the majority of tumor killing by TIL occurs very early after the initiation of therapy. Clin Cancer Res; 1–7. ©2016 AACR.

Thursday, October 6, 2016 6:21 AM|Kohei Shitara, Kimio Yonesaka, Tadamichi Denda, Kentaro Yamazaki, Toshikazu Moriwaki, Masahiro Tsuda, Toshimi Takano, Hiroyuki Okuda, Tomohiro Nishina, Kazuko Sakai, Kazuto Nishio, Shoji Tokunaga, Takeharu Yamanaka, Narikazu Boku, Ichinosuke Hyodo, Kei Muro|Cancer Science|Labels: BRAF, EGFR, HDAC, RAS, VEGF, CRC, clinical trial
This randomized phase II trial compared panitumumab plus fluorouracil, leucovorin, and irinotecan (FOLFIRI) with bevacizumab plus FOLFIRI as second-line chemotherapy for wild-type (WT) KRAS exon 2 metastatic colorectal cancer (mCRC) and to explore the values of oncogenes in circulating tumor DNA (ctDNA) and serum proteins as predictive biomarkers. Patients with WT KRAS exon 2 mCRC refractory to first-line chemotherapy containing oxaliplatin and bevacizumab were randomly assigned to panitumumab plus FOLFIRI or bevacizumab plus FOLFIRI. Of 121 randomly assigned patients, 117 were eligible. Median overall survival (OS) for panitumumab plus FOLFIRI and bevacizumab plus FOLFIRI were 16.2 months and 13.4 months [hazard ratio (HR), 1.16; 95% CI, 0.76–1.77], respectively. Progression-free survival (PFS) was also similar (HR, 1.14; 95% CI, 0.78–1.66). KRAS, NRAS, and BRAF status using ctDNA was successfully examined in 109 patients, and mutations were identified in 19 patients (17.4%). Panitumumab plus FOLFIRI showed favorable survival compared with bevacizumab plus FOLFIRI in WT patients and unfavorable survival in those with mutations (P for interaction = .026 in OS and .054 in PFS). OS with bevacizumab plus FOLFIRI was better than panitumumab plus FOLFIRI in patients with high serum vascular endothelial growth factor-A (VEGF-A) levels and worse in those with low levels (P for interaction = .016). Second-line FOLFIRI plus panitumumab and FOLFIRI plus bevacizumab showed a similar efficacy in patients with WT KRAS exon 2 mCRC. RAS and BRAF mutation in ctDNA could be a negative predictive marker for panitumumab. This article is protected by copyright. All rights reserved.
Wednesday, October 5, 2016 11:19 AM|Lassaletta, Scheinemann, Zelcer, Hukin, Wilson, Jabado, Carret, Lafay-Cousin, Larouche, Hawkins, Pond, Poskitt, Keene, Johnston, Eisenstat, Krishnatry, Mistry, Arnoldo, Ramaswamy, Huang, Bartels, Tabori, Bouffet|Journal of Clinical Oncology Current Issue|Labels: BRAF, brain cancer, clinical trial
Purpose

Vinblastine monotherapy has shown promising activity and a low-toxicity profile in patients with pediatric low-grade glioma (PLGG) who experienced treatment failure after initial treatment with chemotherapy and/or radiation. The aim of this study was to assess the activity of vinblastine in therapy-naïve children.

Patients and Methods

Patients < 18 years old with unresectable and/or progressive therapy-naïve PLGG were eligible. Vinblastine was administered once per week at a dose of 6 mg/m2 intravenously over a period of 70 weeks. Vision, quality of life, neurofibromatosis type 1 (NF1) status, and BRAF mutation/fusion status were also determined and correlated with outcome.

Results

Fifty-four patients were enrolled onto the study, with a median age of 8 years (range, 0.7 to 17.2 years). Most patients had chiasmatic/hypothalamic tumors (55.5%), and 13 patients (24.1%) had NF1. The most common histology was pilocytic astrocytoma (46.3%). Seventeen patients were diagnosed using radiologic criteria alone. Best response to chemotherapy was centrally reviewed with a response rate (complete, partial, or minor response) of 25.9%. Disease stabilization (complete, partial, or minor response or stable disease) was achieved in 47 patients (87.0%). Visual improvement was observed in 20% of patients with optic pathway glioma. Five-year overall survival and progression-free survival (PFS) rates were 94.4% (95% CI, 88.5% to 100%) and 53.2% (95% CI, 41.3% to 68.5%), respectively, for the entire cohort. Patients with NF1 had a significantly better PFS (85.1%; 95% CI, 68.0% to 100%) when compared with patients without NF1 (42.0%; 95% CI, 29.1% to 60.7%; P = .012). Age< 3 years or > 10 years was not associated with poor outcome. Treatment was well tolerated, and quality of life was not affected during treatment. In this trial, there was no correlation between BRAF alterations and outcome.

Conclusion

Vinblastine administered once per week is well tolerated in children with treatment naïve PLGG. Overall survival and PFS are comparable to current therapies, with a favorable toxicity profile and a maintained quality of life.

Monday, October 3, 2016 12:05 AM|Qin, Y., Roszik, J., Chattopadhyay, C., Hashimoto, Y., Liu, C., Cooper, Z. A., Wargo, J. A., Hwu, P., Ekmekcioglu, S., Grimm, E. A.|Molecular Cancer Therapeutics current issue|Labels: BRAF, melanoma, skin cancer

Melanoma is molecularly and structurally heterogeneous, with some tumor cells existing under hypoxic conditions. Our cell growth assays showed that under controlled hypoxic conditions, BRAF(V600E) melanoma cells rapidly became resistant to vemurafenib. By employing both a three-dimensional (3D) spheroid model and a two-dimensional (2D) hypoxic culture system to model hypoxia in vivo, we identified upregulation of HGF/MET signaling as a major mechanism associated with vemurafenib resistance as compared with 2D standard tissue culture in ambient air. We further confirmed that the upregulation of HGF/MET signaling was evident in drug-resistant melanoma patient tissues and mouse xenografts. Pharmacologic inhibition of the c-Met/Akt pathway restored the sensitivity of melanoma spheroids or 2D hypoxic cultures to vemurafenib. Mol Cancer Ther; 15(10); 2442–54. ©2016 AACR.

Monday, October 3, 2016 12:05 AM|Unknown Author|Cancer Discovery current issue|Labels: BRAF, thymic

A new study shows that the BRAF inhibitor vemurafenib may induce partial responses and stable disease in patients with papillary thyroid cancer that has become refractory to radioactive iodine. However, the drug led to a variety of serious adverse events in roughly 65% of patients, including the development of some squamous cell carcinomas.

Monday, October 3, 2016 12:05 AM|Unknown Author|Cancer Discovery current issue|Labels: BRAF, melanoma, skin cancer

STAG2/3 increase sensitivity to BRAF inhibition by reducing ERK activation in melanoma cells.

Saturday, October 1, 2016 1:00 AM|Farchoukh, Lama; Kuan, Shih-Fan; Dudley, Beth; Brand, Randall; Nikiforova, Marina; Pai, Reetesh K.|The American Journal of Surgical Pathology - Current Issue|Labels: BRAF
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.
Saturday, October 1, 2016 1:00 AM|Tsai, Jia-Huei; Liau, Jau-Yu; Yuan, Chang-Tsu; Lin, Yu-Lin; Tseng, Li-Hui; Cheng, Mei-Ling; Jeng, Yung-Ming|The American Journal of Surgical Pathology - Current Issue|Labels: BRAF, WNT
imageRNF43 is an E3 ligase that suppresses the Wnt/β-catenin signaling pathway and is frequently mutated in microsatellite-unstable colorectal carcinoma. To investigate the pathogenetic role of RNF43 in the serrated pathway, we conducted mutation analysis of RNF43 in several types of colorectal neoplasms. RNF43 mutation was found in 2 of 20 (10%) sessile serrated adenomas, 10 of 36 (28%) traditional serrated adenomas, 7 of 37 (19%) traditional serrated adenomas with cytologic dysplasia, and 9 of 31 (29%) BRAF-mutated/microsatellite-stable colorectal carcinomas; however, no mutation was found in 30 tubulovillous/villous adenomas. All mutations were located upstream of the ring finger domain of RNF43 without clustering, which is distinct from the pattern described for microsatellite-unstable colorectal carcinoma. RNF43 mutation was closely associated with BRAF mutation but inversely associated with KRAS mutation in traditional serrated adenoma with or without cytologic dysplasia (P=0.018 and 0.045, respectively). The finding of RNF43 mutation in sessile serrated adenoma and traditional serrated adenoma, but not in tubulovillous/villous adenoma, indicated that RNF43 mutation is an early and specific molecular aberration in the serrated pathway. The frequency of RNF43 mutation was significantly higher in traditional serrated adenoma with or without cytologic dysplasia and BRAF-mutated/microsatellite-stable colorectal carcinoma than sessile serrated adenoma. The unique molecular spectrum of these tumors suggests a stepwise neoplastic progression from sessile serrated adenoma to traditional serrated adenoma and BRAF-mutated/microsatellite-stable colorectal carcinoma, which should be recognized as the traditional serrated pathway to distinguish from the sessile serrated pathway.
Monday, September 26, 2016 2:21 PM|Olow, A., Mueller, S., Yang, X., Hashizume, R., Meyerowitz, J., Weiss, W., Resnick, A. C., Waanders, A. J., Stalpers, L. J. A., Berger, M. S., Gupta, N., James, C. D., Petritsch, C. K., Haas-Kogan, D. A.|Clinical Cancer Research Online First Articles|Labels: BRAF, MapK, mTOR, brain cancer, clinical trial

Purpose: Alteration of the BRAF/MEK/MAPK pathway is the hallmark of pediatric low-grade gliomas (PLGGs), and mTOR activation has been documented in the majority of these tumors. We investigated combinations of MEK1/2, BRAFV600E and mTOR inhibitors in gliomas carrying specific genetic alterations of the MAPK pathway.

Experimental Design: We used human glioma lines containing BRAFV600E (adult high-grade: AM-38, DBTRG, PLGG: BT40), or wild-type BRAF (pediatric high-grade: SF188, SF9427, SF8628) and isogenic systems of KIAA1549:BRAF-expressing NIH/3T3 cells and BRAFV600E-expressing murine brain cells. Signaling inhibitors included everolimus (mTOR), PLX4720 (BRAFV600E), and AZD6244 (MEK1/2). Proliferation was determined using ATP-based assays. In vivo inhibitor activities were assessed in the BT40 PLGG xenograft model.

Results: In BRAFV600E cells, the three possible doublet combinations of AZD6244, everolimus, and PLX4720 exhibited significantly greater effects on cell viability. In BRAFWT cells, everolimus + AZD6244 was superior compared with respective monotherapies. Similar results were found using isogenic murine cells. In KIAA1549:BRAF cells, MEK1/2 inhibition reduced cell viability and S-phase content, effects that were modestly augmented by mTOR inhibition. In vivo experiments in the BRAFV600E pediatric xenograft model BT40 showed the greatest survival advantage in mice treated with AZD6244 + PLX4720 (P < 0.01).

Conclusions: In BRAFV600E tumors, combination of AZD6244 + PLX4720 is superior to monotherapy and to other combinatorial approaches. In BRAFWT pediatric gliomas, everolimus + AZD6244 is superior to either agent alone. KIAA1549:BRAF-expressing tumors display marked sensitivity to MEK1/2 inhibition. Application of these results to PLGG treatment must be exercised with caution because the dearth of PLGG models necessitated only a single patient-derived PLGG (BT40) in this study. Clin Cancer Res; 1–10. ©2016 AACR.

Thursday, September 15, 2016 9:42 AM|Dalin, M. G., Desrichard, A., Katabi, N., Makarov, V., Walsh, L. A., Lee, K.-W., Wang, Q., Armenia, J., West, L., Dogan, S., Wang, L., Ramaswami, D., Ho, A. L., Ganly, I., Solit, D. B., Berger, M. F., Schultz, N. D., Reis-Filho, J. S., Chan, T. A., Morris, L. G. T.|Clinical Cancer Research recent issues|Labels: BRAF, MapK, breast cancer, salivary duct cancer

Purpose: Salivary duct carcinoma (SDC) is an aggressive salivary malignancy, which is resistant to chemotherapy and has high mortality rates. We investigated the molecular landscape of SDC, focusing on genetic alterations and gene expression profiles.

Experimental Design: We performed whole-exome sequencing, RNA sequencing, and immunohistochemical analyses in 16 SDC tumors and examined selected alterations via targeted sequencing of 410 genes in a second cohort of 15 SDCs.

Results: SDCs harbored a higher mutational burden than many other salivary carcinomas (1.7 mutations/Mb). The most frequent genetic alterations were mutations in TP53 (55%), HRAS (23%), PIK3CA (23%), and amplification of ERBB2 (35%). Most (74%) tumors had alterations in either MAPK (BRAF/HRAS/NF1) genes or ERBB2. Potentially targetable alterations based on supportive clinical evidence were present in 61% of tumors. Androgen receptor (AR) was overexpressed in 75%; several potential resistance mechanisms to androgen deprivation therapy (ADT) were identified, including the AR-V7 splice variant (present in 50%, often at low ratios compared with full-length AR) and FOXA1 mutations (10%). Consensus clustering and pathway analyses in transcriptome data revealed striking similarities between SDC and molecular apocrine breast cancer.

Conclusions: This study illuminates the landscape of genetic alterations and gene expression programs in SDC, identifying numerous molecular targets and potential determinants of response to AR antagonism. This has relevance for emerging clinical studies of ADT and other targeted therapies in SDC. The similarities between SDC and apocrine breast cancer indicate that clinical data in breast cancer may generate useful hypotheses for SDC. Clin Cancer Res; 22(18); 4623–33. ©2016 AACR.

Thursday, September 15, 2016 8:45 AM|Teh, J. L. F., Purwin, T. J., Greenawalt, E. J., Chervoneva, I., Goldberg, A., Davies, M. A., Aplin, A. E.|Cancer Research recent issues|Labels: BRAF, CDK, melanoma, skin cancer, regulatory
Aberrant cell-cycle progression is a hallmark feature of cancer cells. Cyclin-dependent kinases 4 and 6 (CDK4/6) drive progression through the G1 stage of the cell cycle, at least in part, by inactivating the tumor suppressor, retinoblastoma. CDK4/6 are targetable and the selective CDK4/6 inhibitor, palbociclib, was recently FDA approved for the treatment of estrogen receptor–positive, HER2-negative advanced breast cancer. In cutaneous melanoma, driver mutations in NRAS and BRAF promote CDK4/6 activation, suggesting that inhibitors such as palbociclib are likely to provide therapeutic benefit in combination with BRAF inhibitors and/or MEK inhibitors that are FDA-approved. However, the determinants of the response to CDK4/6 inhibitors alone and in combination with other targeted inhibitors are poorly defined. Furthermore, in vivo systems to quantitatively and temporally measure the efficacy of CDK4/6 inhibitors and determine the extent that CDK activity is reactivated during acquired resistance are lacking. Here, we describe the heterogeneous effects of CDK4/6 inhibitors, the expression of antiapoptotic proteins that associate with response to CDK4/6 and MEK inhibitors, and the development of a luciferase-based reporter system to determine the effects of CDK4/6 inhibitors alone and in combination with MEK inhibitors in melanoma xenografts. These findings are likely to inform on-going and future clinical trials utilizing CDK4/6 inhibitors in cutaneous melanoma. Cancer Res; 76(18); 5455–66. ©2016 AACR.
Wednesday, September 14, 2016 9:55 AM|PIKOULIS, E., MARGONIS, G. A., ANDREATOS, N., SASAKI, K., ANGELOU, A., POLYCHRONIDIS, G., PIKOULI, A., RIZA, E., PAWLIK, T. M., ANTONIOU, E.|Anticancer Research recent issues|Labels: BRAF, CRC

Background/Aims: The impact of tumor biology on prognosis in patients with colorectal liver metastasis (CRLM) has been the topic of intense research. Specifically, the presence of BRAF mutations has been recently associated with adverse long-term outcomes. We examined the existing literature on the prognostic implications of BRAF mutations in patients with CRLM. Materials and Methods: A structured review of the literature was performed between 5/1/2016 and 6/1/2016 using the PubMed database. Original research articles published between 1/1/2010 and 4/01/2016 were considered eligible. The primary end-points were overall survival (OS)/disease-specific survival (DSS) and recurrence-free survival (RFS) among patients with BRAF mutated CRLM who underwent resection. Results: Eight studies were included. All studies reported on OS/DSS, while 6 reported on RFS. BRAF mutant status was a strong independent predictor of both worse OS/DSS and RFS in 7 and 4 studies, respectively. Conclusion: BRAF-mutant lesions are consistently associated with poor prognosis. Consequently, the indications of CRLM resection in this patient group should be reconsidered.

Wednesday, September 14, 2016 12:05 AM|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.

Wednesday, September 14, 2016 12:05 AM|Fatkhutdinov, N., Sproesser, K., Krepler, C., Liu, Q., Brafford, P. A., Herlyn, M., Aird, K. M., Zhang, R.|Molecular Cancer Research recent issues|Labels: BRAF, melanoma, skin cancer, biomarker diagnostic, regulatory

The majority of patients with melanoma harbor mutations in the BRAF oncogene, thus making it a clinically relevant target. However, response to mutant BRAF inhibitors (BRAFi) is relatively short-lived with progression-free survival of only 6 to 7 months. Previously, we reported high expression of ribonucleotide reductase M2 (RRM2), which is rate-limiting for de novo dNTP synthesis, as a poor prognostic factor in patients with mutant BRAF melanoma. In this study, the notion that targeting de novo dNTP synthesis through knockdown of RRM2 could prolong the response of melanoma cells to BRAFi was investigated. Knockdown of RRM2 in combination with the mutant BRAFi PLX4720 (an analog of the FDA-approved drug vemurafenib) inhibited melanoma cell proliferation to a greater extent than either treatment alone. This occurred in vitro in multiple mutant BRAF cell lines and in a novel patient-derived xenograft (PDX) model system. Mechanistically, the combination increased DNA damage accumulation, which correlated with a global decrease in DNA damage repair (DDR) gene expression and increased apoptotic markers. After discontinuing PLX4720 treatment, cells showed marked recurrence. However, knockdown of RRM2 attenuated this rebound growth both in vitro and in vivo, which correlated with maintenance of the senescence-associated cell-cycle arrest.

Implications: Inhibition of RRM2 converts the transient response of melanoma cells to BRAFi to a stable response and may be a novel combinatorial strategy to prolong therapeutic response of patients with melanoma. Mol Cancer Res; 14(9); 767–75. ©2016 AACR.

Friday, September 2, 2016 12:05 AM|Theurich, S., Rothschild, S. I., Hoffmann, M., Fabri, M., Sommer, A., Garcia-Marquez, M., Thelen, M., Schill, C., Merki, R., Schmid, T., Koeberle, D., Zippelius, A., Baues, C., Mauch, C., Tigges, C., Kreuter, A., Borggrefe, J., von Bergwelt-Baildon, M., Schlaak, M.|Cancer Immunology Research recent issues|Labels: BRAF, melanoma, skin cancer

Immune checkpoint inhibition with ipilimumab has revolutionized cancer immunotherapy and significantly improved outcomes of patients with advanced malignant melanoma. Local peripheral treatments (LPT), such as radiotherapy or electrochemotherapy, have been shown to modulate systemic immune responses, and preliminary data have raised the hypothesis that the combination of LPT with systemic immune checkpoint blockade might be beneficial. Clinical data from 127 consecutively treated melanoma patients at four cancer centers in Germany and Switzerland were analyzed. Patients received either ipilimumab (n = 82) or ipilimumab and additional LPT (n = 45) if indicated for local tumor control. The addition of LPT to ipilimumab significantly prolonged overall survival (OS; median OS 93 vs. 42 weeks, unadjusted HR, 0.46; P = 0.0028). Adverse immune-related events were not increased by the combination treatment, and LPT-induced local toxicities were in most cases mild. In a multivariable Cox regression analysis, we show that the effect of added LPT on OS remained statistically significant after adjusting for BRAF status, tumor stage, tumor burden, and central nervous system metastases (adjusted HR, 0.56; 95% confidence interval, 0.31–1.01, P = 0.05). Our data suggest that the addition of LPT to ipilimumab is safe and effective in patients with metastatic melanoma irrespective of clinical disease characteristics and known risk factors. Induction of antitumor immune responses is most likely the underlying mechanism and warrants prospective validation. Cancer Immunol Res; 4(9); 744–54. ©2016 AACR.

Thursday, September 1, 2016 12:05 AM|Pietrantonio, F., Oddo, D., Gloghini, A., Valtorta, E., Berenato, R., Barault, L., Caporale, M., Busico, A., Morano, F., Gualeni, A. V., Alessi, A., Siravegna, G., Perrone, F., Di Bartolomeo, M., Bardelli, A., de Braud, F., Di Nicolantonio, F.|Cancer Discovery recent issues|Labels: BRAF, EGFR, CRC

A patient with metastatic BRAF-mutated colorectal cancer initially responded to combined EGFR and BRAF inhibition with panitumumab plus vemurafenib. Pre-existing cells with increased MET gene copy number in the archival tumor tissue likely underwent clonal expansion during treatment, leading to the emergence of MET amplification in the rebiopsy taken at progression. In BRAF-mutated colorectal cancer cells, ectopic expression of MET conferred resistance to panitumumab and vemurafenib, which was overcome by combining BRAF and MET inhibition. Based on tumor genotyping and functional in vitro data, the patient was treated with the dual ALK–MET inhibitor crizotinib plus vemurafenib, thus switching to dual MET and BRAF blockade, with rapid and marked effectiveness of such strategy. Although acquired resistance is a major limitation to the clinical efficacy of anticancer agents, the identification of molecular targets emerging during the first treatment may afford the opportunity to design the next line of targeted therapies, maximizing patient benefit.

Significance: MET amplification is here identified—clinically and preclinically—as a new mechanism of resistance to EGFR and BRAF dual/triple block combinations in BRAF-mutated colorectal cancer. Switching from EGFR to MET inhibition, while maintaining BRAF inhibition, resulted in clinical benefit after the occurrence of MET-driven acquired resistance. Cancer Discov; 6(9); 963–71. ©2016 AACR.

This article is highlighted in the In This Issue feature, p. 932

Wednesday, August 31, 2016 8:00 PM|Grothey A, Sargent DJ. |JAMA Oncology Current Issue|Labels: BRAF, EGFR, HDAC, CRC
In current practice, the choice of which adjuvant therapy to use for colon cancer is determined mainly on the basis of the patient’s TNM stage. Patients with stage III cancers are most commonly considered candidates for a fluoropyrimidine plus oxaliplatin treatment; those with stage II cancers, for a fluoropyrimidine alone or no postsurgical therapy. This standard of care has not changed for more than a decade. With the exception of high microsatellite instability (MSI-H) status (which characterizes a subgroup of 15% to 20% of patients with stage II disease who have excellent prognosis and thus no need for adjuvant therapy), no molecular markers or marker signatures have made relevant inroads into clinical practice. Commercial tests using a limited gene-expression profile analysis are currently being marketed as decision tools for the use of adjuvant therapy in patients with stage II colon cancer, but since these tests have shown only prognostic and not predictive value, their utility in clinical practice is limited. This is in contrast to the situation in metastatic colorectal cancer, for which biomarkers are increasingly being used to guide treatment decisions in clinical practice. Activating mutations in exons 2, 3, and 4 of KRAS and NRAS genes have been established as negative predictive markers for the activity of anti–epidermal growth factor receptors such as cetuximab and panitumumab. Patients whose tumors show BRAF V600E mutations are known to have a very poor prognosis, and intensified chemotherapy regimens as well as combinations of targeted agents are being developed for this specific population. Hypermutated MSI-H cancers have been identified as targets for immune therapy with anti–PD-1/PD-L1 (programmed cell death 1/programmed cell death 1 ligand 1) agents, and human epithelial growth factor receptor 2 (HER2)-positive colorectal cancers appear to respond to HER2-targeted combination therapy. These markers, however, characterize only about 15% to 20% of patients with advanced colorectal cancer, and importantly, they represent only single molecular alterations, which are conceivably not an adequate reflection of the complex biology of a cancer cell.
Wednesday, August 31, 2016 10:36 AM|Immacolata Maida; Stefania Guida; Amalia Azzariti; Letizia Porcelli; Stefania Tommasi; Paola Zanna; Tiziana Cocco; Michele Guida; Gabriella Guida|JournalTOCs API - Biochimica et Biophysica Acta (BBA) - Molecular Cell Research (50 articles)|Labels: BRAF, melanoma, skin cancer

New insight into the role of metabolic reprogramming in melanoma cells harboring BRAF mutations
Anna Ferretta Immacolata Maida; Stefania Guida; Amalia Azzariti; Letizia Porcelli; Stefania Tommasi; Paola Zanna; Tiziana Cocco; Michele Guida; Gabriella Guida
Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, Vol. 1863, No. 11 (2016) pp. 2710 - 2718
Publication date: November 2016 Source:Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, Volume 1863, Issue 11 Author(s): Anna Ferretta, Immacolata Maida, Stefania Guida, Amalia Azzariti, Letizia Porcelli, Stefania Tommasi, Paola Zanna, Tiziana Cocco, Michele Guida, Gabriella Guida This study explores the V600BRAF-MITF-PGC-1α axis and compares metabolic and functional changes occurring in primary and metastatic V600BRAF melanoma cell lines. V600BRAF mutations in homo/heterozygosis were found to be correlated to high levels of pERK, to downregulate PGC-1α/β, MITF and tyrosinase activity, resulting in a reduced melanin synthesis as compared to BRAFwt melanoma cells. In this scenario, V600BRAF switches on a metabolic reprogramming in melanoma, leading to a decreased OXPHOS activity and increased glycolytic ATP, lactate, HIF-1α and MCT4 levels. Furthermore, the induction of autophagy and the presence of ER stress markers in V600BRAF metastatic melanoma cells suggest that metabolic adaptations of these cells occur as compensatory survival mechanisms. For the first time, we underline the role of peIF2α as an important marker of metastatic behaviour in melanoma. Our results suggest the hypothesis that inhibition of the glycolytic pathway, inactivation of peIF2α and a reduction of basal autophagy could be suitable targets for novel combination therapies in a specific subgroup of metastatic melanoma.

Monday, August 15, 2016 12:05 AM|Harbst, K., Lauss, M., Cirenaȷwis, H., Isaksson, K., Rosengren, F., Torngren, T., Kvist, A., Johansson, M. C., Vallon–Christersson, J., Baldetorp, B., Borg, A., Olsson, H., Ingvar, C., Carneiro, A., Jonsson, G.|Cancer Research recent issues|Labels: BRAF, melanoma, skin cancer
Cancer genome sequencing has shed light on the underlying genetic aberrations that drive tumorigenesis. However, current sequencing-based strategies, which focus on a single tumor biopsy, fail to take into account intratumoral heterogeneity. To address this challenge and elucidate the evolutionary history of melanoma, we performed whole-exome and transcriptome sequencing of 41 multiple melanoma biopsies from eight individual tumors. This approach revealed heterogeneous somatic mutations in the range of 3%–38% in individual tumors. Known mutations in melanoma drivers BRAF and NRAS were always ubiquitous events. Using RNA sequencing, we found that the majority of mutations were not expressed or were expressed at very low levels, and preferential expression of a particular mutated allele did not occur frequently. In addition, we found that the proportion of ultraviolet B (UVB) radiation-induced C>T transitions differed significantly (P < 0.001) between early and late mutation acquisition, suggesting that different mutational processes operate during the evolution of metastatic melanoma. Finally, clinical history reports revealed that patients harboring a high degree of mutational heterogeneity were associated with more aggressive disease progression. In conclusion, our multiregion tumor-sequencing approach highlights the genetic evolution and non-UVB mutational signatures associated with melanoma development and progression, and may provide a more comprehensive perspective of patient outcome. Cancer Res; 76(16); 4765–74. ©2016 AACR.
Monday, August 15, 2016 12:05 AM|Bossi, D., Cicalese, A., Dellino, I. G., Luzi, L., Punzi, S., D'Alesio, C., Cavallaro, E., Minucci, S., Pelicci, P., Lanfrancone, L.|Clinical Cancer Research recent issues|Labels: BRAF, melanoma, skin cancer, regulatory

Metastatic melanoma is one of the most aggressive cancers, refractory to most of the current therapies in its late stage. Melanoma is characterized by a strong molecular heterogeneity, and patients can be stratified in different subtypes according to genetic alterations. Despite the approval of new targeted drugs, patient treatment produces only partial responses and frequent and rapid relapses, mostly due to increased resistance. Mounting evidence highlights the necessity to better stratify melanomas on the basis of altered molecular pathways and to define new specific and effective tailored therapies. Our main goal is the genetic and functional characterization of metastatic melanomas aimed to generate new predictive and prognostic biomarkers and to identify new potential "druggable" candidates involved in melanoma progression.

We have generated a series of human-in-mouse melanoma models, by xeno-transplantation of a cohort of human melanoma samples in immune-compromised mice. Our patient-derived xenografts (PDXs) faithfully recapitulate the heterogeneity of the original tumors at phenotypic and genetic level. Taking advantage of our model, we performed the first in vivo genetic screen on patient-derived tumors, using metastatic NRAS- and BRAF-mutant melanomas and targeting chromatin genes (~240). Our screens revealed unprecedented numerosity of tumor drivers (~50% of tested genes) and unexpected functional heterogeneity among patients (~60 drivers per tumor; <15% in common). Tumor drivers, however, were not activated by somatic mutations in the same patients. Several genes have been validated in vivo and then characterized in vitro on primary cultures derived from the patients' cells. We analysed underlying molecular mechanisms of one of the identified drivers, the Histone-lysine N-methyltransferase KMT2D, and showed that it promotes tumorigenesis by dysregulating a subset of transcriptional enhancers and genes involved in cell migration.

Citation Format: Daniela Bossi, Angelo Cicalese, Ivan Gaetano Dellino, Lucilla Luzi, Simona Punzi, Carolina D'Alesio, Elena Cavallaro, Saverio Minucci, PierGiuseppe Pelicci, Luisa Lanfrancone. Challenging new epigenetic vulnerabilities in human metastatic melanoma PDXs. [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 A28.

Monday, August 15, 2016 12:05 AM|Houghton, P. J., Kurmasheva, R. T.|Clinical Cancer Research recent issues|Labels: BRAF, biomarker diagnostic

The Pediatric Preclinical Testing Program (PPTP) has evaluated almost 97 single agents and several combinations over the last 10 years. A total of 89 xenograft models that encompass solid tumors, brain tumors, and acute leukemias have been used as a screen to identify novel agents that could be accelerated for clinical trial. With few exceptions, all models were directly established in mice from patient biopsies, and approximately half of the models were established from patients who were refractory or had relapsed.

Expression analysis showed that tumor models clustered with their appropriate clinical histotype, and exome sequencing showed mutation frequency and identity to be similar to published genomic data for patient cancers. The models accurately identified known active agents that are used as standard of care therapy for solid tumors and leukemias, thus validating the models.

Criteria for assessing response similar to clinical criteria were developed. Of the 97 agents tested, twenty-four signaling inhibitors (ligand or receptor binding antibodies, kinase inhibitors etc.) were evaluated. With the exception of inhibitors of Aurora kinase A, and Polo-like kinase 1, the overall objective response rate (ORR) was <2%. The low response rate is probably an accurate reflection of likely ORR in clinical trials, as the models identified selumetinib in an astrocytoma with BRAF (V600E) mutation, dasatinib in a Ph+ ALL, sorafenib in a Flt3 mutant ALL, and crizotinib in an ALK mutant neuroblastoma.

The models identified several cytotoxic agents as having broad-spectrum antitumor activity (eribulin, alisertib, PR-104), with small molecule and antibody conjugated antimitotic agents showing high-level activity in many models (eribulin in sarcoma and ALL, glembatumab in osteosarcoma). The major issue is that the models overpredict drug activity due to tolerance of the host (mouse) compared to patients. Thus, it is imperative to adjust dose/schedule to accurately reproduce clinical exposures.

Another issue to be addressed is whether the models adequately encompass genomic and epigenomic heterogeneity. With increasing evidence of molecular subtypes within histotypes previously considered as relatively homogeneous, it is clear that additional models need to be established. We have analyzed 2134 tumor/drug studies undertaken as part of the PPTP to determine whether alternative experimental designs can be used that would allow an increased genomic diversity to be interrogated using current resources.

Citation Format: Peter J. Houghton, Raushan T. Kurmasheva. Promise and challenges of pediatric cancer PDX models. [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 IA15.

Monday, August 15, 2016 12:05 AM|Banda, M., McKim, K. L., Parsons, B.|Clinical Cancer Research recent issues|Labels: BRAF, EGFR, lung cancer

Molecularly-targeted therapeutics are being developed for personalized cancer treatment based on the ideas that treatments should be determined by specific characteristics of an individual patient's tumor and that treatments should target tumor cells without damaging normal cells. Accumulating evidence indicates tumor heterogeneity can lead to acquired resistance to molecularly-targeted therapies. Currently, preclinical investigations rely heavily on the use of tumor cell lines and mouse xenograft models. The high failure rate in oncology drug development suggests that such test systems do not adequately model tumor heterogeneity. Therefore, more predictive preclinical models are needed to ensure that only the most promising molecularly-targeted, combination therapies are assessed in clinical studies. Here we established a novel 3D lung adenocarcinoma tissue-originating spheroid model, which may be used to investigate the efficacy of molecularly-targeted cancer therapeutics. Fresh lung adenocarcinomas were minced and either digested with Liberase DH enzyme (5.6 units) or homogenized to generate cancer tissue-originating spheroids. Spheroids were plated in Matrigel and cultured in stem cell media, with 0.01, 0.1, 1 or 10 μM erlotinib or the DMSO vehicle. Erlotinib is an epidermal growth receptor inhibitor commonly used to treat non-small cell lung cancer (NSCLC). Using an ImageXpress Micro Widefield High Content Screening System (Molecular devices), z-stack bright field images were collected 1, 7, and 14 days after plating. Composite z-stack images were analyzed using LASX (Leica Microsystems) software. Total spheroid area (in pixels) was measured for replicate cultures exposed to different erlotinib concentrations at each time point. The total area of spheroids for 7 and 14 days were normalized to day 1 measurements and cell death was evaluated as the decrease in total area of the spheroids in culture. Our results indicate that direct homogenization of lung adenocarcinomas generated at least ten times greater yield of spheroids than was achieved using digestion with the Liberase DH. The yield and appearance of spheroids varied significantly. Spheroids prepared from some lung adenocarcinomas were more spherical and aggregated than those prepared from other tumors. There was time-dependent decrease in total area of spheroids following 7 and 14 days in culture. Also spheroids prepared from different lung adenocarcinomas responded differently to different concentrations of erlotinib. High-sensitivity quantification of specific mutations (EGFR, KRAS PIK3CA and BRAF) will be conducted before and after erlotinib treatments to determine whether this model can be used to assess the potential outgrowth of drug-resistant tumor subpopulations.

Citation Format: Malathi Banda, Karen L. McKim, Barbara Parsons. Establishing an ex-vivo, tumor spheroid culture system to assess molecular-targeted therapies for personalized treatment of non-small cell lung cancer. [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 A15.

Monday, August 15, 2016 12:05 AM|Kopetz, S.|Clinical Cancer Research recent issues|Labels: BRAF, EGFR, biomarker diagnostic

Patient-derived xenograft models (PDX) provide opportunities to evaluate efficacy, pharmacodynamics, response durability, and resistance mechanisms in greater details than traditionally possible with sampling of patient tumors. When PDX models are integrated with patients enrolled in clinical trials, or treated with novel agents, there are opportunities to interrogate aspects of cancer biology not traditionally evaluable, while maintaining clinical relevance. This presentation will explore the strengths and limitations of the models, with examples on how these models are being integrated into clinical trials, validation efforts, and novel insights derived from this approach.

The concept of a co-clinical trial has been variously defined, including a definition proposed by the NCI as parallel or sequential trials of combination therapy in patients and in mouse and human-in-mouse models of appropriate genotypes to represent the patients. This concept is being extended for PDX models to incorporate tumor models derived from the same patients participating in the clinical trial, both with a goal of improving translational interrogation of the novel agent efficacy as well as emerging efforts to use the PDX models to guide patient therapy.

Validation of the PDX models for treatment response/regression will be demonstrated, including recently completed clinical trials utilizing inhibitors of BRAF and MEK with a comparison of the response of the matched PDX models to radiographic measurements from the treated patients. Cohort based validation will be demonstrated through examples with EGFR targeted therapy and comparison to randomized clinical trial results. Limitations of these approaches will be described.

PDX models from clinical trials have been demonstrated to model and replicate mechanisms of resistance in some settings, and examples from colorectal and lung cancer will be used to demonstrate this. Resistance of lung cancer to EGFR tyrosine kinase inhibitors is being explored on PDXs obtained from post-progression biopsies. Tumor heterogeneity as a mechanism of resistance has been successfully modeled in BRAF-mutant colorectal cancer, and has been integrated into prospective clinical trials.

Finally, efforts to integrate PDX models as predictors of treatment response are described. These include efforts to use matched PDX models to guide treatment decisions in patients as part of prospective clinical trials. Regulatory, logistic, and biologic limitations of such efforts will be discussed.

Citation Format: Scott Kopetz. Opportunities for integration of PDX models into clinical trials. [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 IA21.

Monday, August 15, 2016 12:05 AM|Krepler, C., Sproesser, K., Beqiri, M., Xiao, M., Brafford, P., Xu, W., Garman, B., Wargo, J., Davies, M. A., Frederick, D. T., Flaherty, K. T., Hoon, D., Bennett, J. J., Guarino, M., Petrelli, N. J., Elder, D., Xu, X., Karakousis, G., NATHANSON, K. L., Schuchter, L., Herlyn, M.|Clinical Cancer Research recent issues|Labels: BRAF, melanoma, skin cancer, biomarker diagnostic

Advanced melanoma has seen dramatic changes in standard of care in the last years and many novel targeted small molecules and immune checkpoint inhibitors are in development. More than 200 clinical trials are currently ongoing for metastatic melanoma. Thus, accurate pre-clinical models to predict clinical responses are urgently needed. We have established a large bank of live tumor tissue (n=500) with more than 300 models expanded as PDX. Melanoma tumor tissue is uniquely suited to establish patient-derived xenograft (PDX) models, since only one tumor cell can initiate tumor growth. Thus, our success rate is at 90% using NSG mice and matrigel for s.c. implantation of minced tissue chunks. We have opted for such a large number of models due to the genomic and clinical heterogeneity of melanoma based on available TCGA data. Although almost half of all melanomas harbor BRAF V600 hotspot mutations, followed in frequency by NRAS and NF1 mutations, many driver mutations are only found in a small subset and many mutations are occurring concurrently in various confirmations. We used a custom targeted capture of 108 genes previously implicated in melanoma, and performed massively parallel sequencing on currently 200 PDX. Samples were then clustered into groups based on deleterious mutations which were detected in all but a very small subset of samples. The biological relevance of these multiple genomic subsets was then tested against latency, growth rate, and spontaneous metastasis in PDX. Further, clinical information such as age, gender, history, and response to therapies provided additional parameters to classify this collection into meaningful subgroups. In conclusion, this collection of melanoma PDX recapitulates the breadth of advanced melanoma in the clinic and therefore a comprehensive resource for precision medicine testing in an increasingly scattered therapy landscape.

Citation Format: Clemens Krepler, Katrin Sproesser, Marilda Beqiri, Min Xiao, Patricia Brafford, Wei Xu, Bradley Garman, Jennifer Wargo, Michael A. Davies, Dennie T. Frederick, Keith T. Flaherty, David Hoon, Joseph J. Bennett, Michael Guarino, Nicholas J. Petrelli, David Elder, Xiaowei Xu, Giorgos Karakousis, KATHERINE L. NATHANSON, Lynn Schuchter, Meenhard Herlyn. A comprehensive collection of patient derived xenografts of human melanoma with clinical, genomic, and biological characterization. [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 A01.

Thursday, August 11, 2016 3:02 PM|Rose, A. A. N., Annis, M. G., Frederick, D. T., Biondini, M., Dong, Z., Kwong, L. N., Chin, L., Keler, T., Hawthorne, T., Watson, I. R., Flaherty, K. T., Siegel, P. M.|Clinical Cancer Research Online First Articles|Labels: BRAF, MapK, melanoma, skin cancer

Purpose: To determine if BRAF and/or MEK inhibitor-induced GPNMB expression renders melanomas sensitive to CDX-011, an antibody-drug conjugate targeting GPNMB. Experimental Design: The TCGA melanoma dataset was interrogated for a panel of MITF-regulated melanosomal differentiation antigens, including GPNMB. BRAF mutant melanoma cell lines treated with BRAF or MEK inhibitors were assessed for GPNMB expression by RT-qPCR, immunoblot and FACs analyses. Transient siRNA-mediated knockdown approaches were used to determine if MITF is requirement for treatment-induced GPNMB upregulation. GPNMB expression was analyzed in serial biopsies and serum samples from melanoma patients taken before, during and after disease progression on MAPK inhibitor treatment. Sub-cutaneous injections were performed to test the efficacy of MAPK inhibitors alone, CDX-011 alone, or their combination in suppressing melanoma growth. Results: A MITF-dependent melanosomal differentiation signature is associated with poor prognosis in patients with this disease. MITF is increased following BRAF and MEK inhibitor treatment and induces the expression of melanosomal differentiation genes, including GPNMB. GPNMB is expressed at the cell surface in MAPK inhibitor-treated melanoma cells and is also elevated in on-treatment versus pre-treatment biopsies from melanoma patients receiving MAPK pathway inhibitors. Combining BRAF and/or MEK inhibitors with CDX-011, an antibody-drug-conjugate targeting GPNMB, is effective in causing melanoma regression in pre-clinical animal models and delays the recurrent melanoma growth observed with MEK or BRAF/MEK inhibitor treatment alone. Conclusions: The combination of MAPK pathway inhibitors with an antibody-drug-conjugate targeting GPNMB is an effective therapeutic option for patients with melanoma.

Thursday, August 4, 2016 8:57 AM|Yang, X., Li, J., Li, X., Liang, Z., Gao, W., Liang, J., Cheng, S., Lin, Y.|JNM Ahead of Print|Labels: BRAF, telomerase

Telomerase Reverse Transcriptase (TERT) promoter mutation has been reported to be associated with aggressive characteristics in differentiated thyroid cancer (DTC). This study examined the status of TERT promoter mutation in distant metastatic DTC (DM-DTC), and evaluated the correlation between TERT mutation and radioactive iodine-131(RAI) uptake, as well as that between TERT mutation and therapy response. Methods: TERT promoter and B-Raf proto-oncogene (BRAF) V600E mutation were retrospectively examined in primary tumors of 66 DM-DTC patients. Stimulated thyroglobulin (sTg) changes, RAI uptake status (avid or non-avid), and other imaging evidence were analyzed to evaluate therapy response. After a median follow-up of 46.5 months (interquartile range, 29.0 to 70.5 months), therapy response was classified as disease control and refractory. Results: The prevalence of TERT mutations was 22.73% (15/66), of which C228T mutation was more prevalent (13/15) than C250T mutation (2/15). Rising sTg was noticed in 93.33% (14/15) of TERT mutation group. While in cases with both mutations negative, 78.12 % (25/32) presented with decreased sTg. TERT mutation closely correlated with poor RAI therapy response (p<0.001), and all 15 patients were classified as refractory to RAI with a positive predictive value of 100% at the end point of follow-up. TERT mutation was associated with older mean age at diagnosis (p<0.001), larger mean tumor diameter (P = 0.013), and more likelihood of both BRAF mutation coexistence (P = 0.044) and refractory to RAI (p<0.001). In the 36 cases received imaging semi-quantitative analysis, it was found that TERT mutation significantly correlated with non-RAI-avidity, with a much lower mean tumor/background (T/B) ratio (obtained from post RAI therapy whole-body scanning) than TERT wild-type (p<0.001). And DM-DTC patients with TERT mutation were more likely to lose RAI-avidity at initial RAI therapy than those with only BRAF mutation (8/8 vs 5/11, Fisher’s exact test, P = 0.018). Conclusion: TERT promoter mutation closely associates with non-RAI-avidity in DM-DTC, and when comparing with BRAF mutation, TERT mutation manifested a worse negative influence on RAI uptake. It could also be used as a predictive marker to early identify refractory to RAI.

Sunday, July 31, 2016 8:00 PM|Falcone, A.|The Oncologist Subject Collection: Gastrointestinal Cancer|Labels: BRAF, EGFR, RAS, CRC
Introduction.Right- and left-sided colorectal cancers (CRCs) differ in clinical and molecular characteristics. Some retrospective analyses suggested that patients with right-sided tumors derive less benefit from anti-epidermal growth factor receptor (EGFR) antibodies; however, molecular selection in those studies was not extensive. Patients and Methods.Patients with RAS and BRAF wild-type metastatic CRC (mCRC) who were treated with single-agent anti-EGFRs or with cetuximab-irinotecan (if refractory to previous irinotecan) were included in the study. Differences in outcome between patients with right- and left-sided tumors were investigated. Results.Of 75 patients, 14 and 61 had right- and left-sided tumors, respectively. None of the right-sided tumors responded according to RECIST, compared with 24 left-sided tumors (overall response rate: 0% vs. 41%; p = .0032), and only 2 patients with right-sided tumors (15%) versus 47 patients with left-sided tumors (80%) achieved disease control (p < .0001). The median duration of progression-free survival was 2.3 and 6.6 months in patients with right-sided and left-sided tumors, respectively (hazard ratio: 3.97; 95% confidence interval: 2.09-7.53; p < .0001). Conclusion.Patients with right-sided RAS and BRAF wild-type mCRC seemed to derive no benefit from single-agent anti-EGFRs. Implications for PracticeRight- and left-sided colorectal tumors have peculiar epidemiological and clinicopathological characteristics, distinct gene expression profiles and genetic alterations, and different prognoses. This study assessed the potential predictive impact of primary tumor site with regard to anti-epidermal growth factor receptor (EGFR) monoclonal antibody treatment in patients with RAS and BRAF wild-type metastatic colorectal cancer. The results demonstrated the lack of activity of anti-EGFRs in RAS and BRAF wild-type, right-sided tumors, thus suggesting a potential role for primary tumor location in driving treatment choices.
Sunday, July 17, 2016 5:00 PM|Pathology International|Pathology International via MedWorm.com|Comments|Labels: BRAF, GIST
Gastrointestinal stromal tumor (GIST) is a mesenchymal tumor of the gastrointestinal tract. Mutation of KIT and PDGFRA genes is implicated in the tumorigenesis. Approximately 10% of GISTs do not harbor mutation of these genes, and they are designated as “wild type” GIST. They are classified into succinate dehydrogenase (SDH)‐deficient and non‐SDH‐deficient groups. SDH‐deficient group includes Carney triad and Carney Stratakis syndrome. The patients are young women. Tumors occur in the antrum of the stomach, and tumor cells are epithelioid. Lymph node metastasis is frequent. The non‐SDH‐deficient group includes neurofibromatosis (NF) type 1 and GISTs with mutations of BRAF, KRAS, and PIK3CA and with the ETV6‐NTRK3 fusion gene. GIST in NF occurs in the small intestine, and ...
Thursday, July 7, 2016 9:46 AM|Gasparotto, D., Rossi, S., Polano, M., Tamborini, E., Lorenzetto, E., Sbaraglia, M., Mondello, A., Massani, M., Lamon, S., Bracci, R., Mandolesi, A., Frate, E., Stanzial, F., Agaj, J., Mazzoleni, G., Pilotti, S., Gronchi, A., Dei Tos, A. P., Maestro, R.|Clinical Cancer Research Online First Articles|Labels: BRAF, GIST

Purpose: The majority of Gastrointestinal Stromal Tumors (GISTs) are driven by KIT, PDGFRA or, less commonly, BRAF mutations, and SDH gene inactivation is involved in a limited fraction of gastric lesions. However, about 10% of GISTs are devoid of any of such alterations and are poorly responsive to standard treatments. This study aims to shed light on the molecular drivers of quadruple-negative GISTs. Experimental Design: Twenty-two sporadic quadruple-negative GISTs with no prior association with Neurofibromatosis Type 1 syndrome were molecularly profiled for a panel of genes belonging to tyrosine kinase pathways or previously implicated in GISTs. For comparison purposes, 24 GISTs carrying KIT, PDGFRA, or SDH gene mutations were also analyzed. Molecular findings were correlated to clinicopathological features. Results: Most quadruple-negative GISTs featured intestinal localization, with a female predilection. About 60% (13/22) of quadruple-negative tumors carried NF1 pathogenic mutations, often associated with biallelic inactivation. The analysis of normal tissues, available in 11 cases, indicated the constitutional nature of the NF1 mutation in 7/11 cases, unveiling an unrecognized Neurofibromatosis Type 1 syndromic condition. Multifocality and a multinodular pattern of growth were common findings in NF1-mutated quadruple-negative GISTs. Conclusions: NF1 gene mutations are frequent in quadruple-negative GISTs and are often constitutional, indicating that a significant fraction of patients with apparently sporadic quadruple-negative GISTs are affected by unrecognized Neurofibromatosis Type 1 syndrome. Hence, a diagnosis of quadruple-negative GIST, especially if multifocal or with a multinodular growth pattern and a non-gastric location, should alert the clinician to a possible Neurofibromatosis Type 1 syndromic condition.

Friday, July 1, 2016 12:40 AM|J.H. Beijnen|JournalTOCs API - Journal of Clinical Pharmacology (94 articles)|Labels: BRAF, melanoma, skin cancer, clinical trial

Clinical Pharmacokinetics of Vemurafenib in BRAF‐Mutated Melanoma Patients
C.M. Nijenhuis A.D.R. Huitema, C. Blank, J.B.A.G. Haanen, J.V. Thienen, H. Rosing, J.H.M. Schellens, J.H. Beijnen
Journal of Clinical Pharmacology, Vol. , No. (2016) pp. -
Vemurafenib is an oral tyrosine kinase inhibitor which inhibits mutated serine/threonine protein kinase B‐Raf (BRAF) and is approved as monotherapy or in combination with the MEK inhibitor cobimetinib for the treatment of adult patients with BRAF V600 mutation‐positive unresectable or metastatic melanoma.1‐3 Currently vemurafenib is given in a fixed dose regimen of 960 mg bi‐daily (BID). The pharmacokinetics of vemurafenib was previously investigated in a phase I trial and mean steady state plasma concentration of 40 ± 20 μg/mL was found at.4 Concomitant intake of food (high‐fat meal) increased the Cmax (2.5 times) and the vemurafenib plasma AUC0‐∞ (4.7 times).5 In addition a dose‐exposure relationship has been established for vemurafenib at steady state from 240 mg BID to 960 mg BID.6 This article is protected by copyright. All rights reserved

Friday, July 1, 2016 12:05 AM|Unknown Author|Cancer Discovery recent issues|Labels: BRAF, clinical trial

In a phase I trial, BGB-283, which targets the RAF family of proteins, was found to be safe and tolerable—and effective—in treating patients with a variety of solid tumors harboring mutations in BRAF, KRAS, and NRAS.

Friday, July 1, 2016 12:05 AM|Unknown Author|Cancer Discovery recent issues|Labels: BRAF, lung cancer, clinical trial

Results from a phase II trial show that the BRAF inhibitor dabrafenib has significant single-agent activity in patients with advanced non–small cell lung cancer harboring the BRAF V600E mutation. However, data from another arm of the study suggest that combining dabrafenib with a MEK inhibitor may be a more effective treatment strategy for these patients.

Monday, June 27, 2016 2:02 PM|Barras, D., Missiaglia, E., Wirapati, P., Sieber, O. M., Jorissen, R. N., Love, C., Molloy, P. L., Jones, I. T., McLaughlin, S., Gibbs, P., Guinney, J., Simon, I. M., Roth, A., Bosman, F. T., Tejpar, S., Delorenzi, M.|Clinical Cancer Research Online First Articles|Labels: BRAF, EGFR, CRC

Purpose: Mutation of BRAF at the valine 600 residue occurs in ~10% of colorectal cancers (CRC), a group with particularly poor prognosis. The response of BRAF mutant CRC to recent targeted strategies such as anti-BRAF or combinations with MEK and EGFR inhibitors remains minimal and highly heterogeneous within BRAF V600E cohorts. There is clearly an unmet need in understanding the biology of BRAF V600E CRC and potential subgroups within this population. Experimental Design: In the biggest yet reported cohort of 218 BRAF V600E with gene expression data, we performed unsupervised clustering using non-negative matrix factorization to identify gene expression based subgroups and characterized pathway activation. Results: We found strong support for a split into two distinct groups, called BM1 and BM2. These subtypes are independent of MSI status, PI3K mutation, gender and sidedness. Pathway analyses revealed that BM1 is characterized by KRAS/AKT pathway activation, mTOR/4EBP deregulation and EMT while BM2 displays important deregulation of the cell cycle. Proteomics data validated these observations as BM1 is characterized by high phosphorylation levels of AKT and 4EBP1, and BM2 patients display high CDK1 and low cyclin D1 levels. We provide a global assessment of gene expression motifs that differentiate BRAF V600E subtypes from other CRC. Conclusions: We suggest that BRAF mutant patients should not be considered as having a unique biology and provide an in depth characterization of heterogeneous motifs that may be exploited for drug targeting.

Monday, June 27, 2016 12:52 PM|LUO, H., UMEBAYASHI, M., DOI, K., MORISAKI, T., SHIRASAWA, S., TSUNODA, T.|Anticancer Research recent issues|Labels: AKT, BRAF, melanoma, skin cancer

Background/Aim: The serine/threonine-protein kinase B-Raf (BRAF) V600E mutant (BRAFV600E) inhibitor vemurafenib, has improved clinical outcomes for patients with BRAFV600E melanoma, but acquired cellular resistance mediated by AKT serine/threonine kinase 1 (AKT) phosphorylation limits its efficacy. We examined the effect of resveratrol on vemurafenib-resistant melanoma cells. Materials and Methods: A vemurafenib-resistant human metastatic melanoma cell line positive for the BRAF V600E mutation was established. The anti-tumorigenic effects of vemurafenib and resveratrol, both alone and in combination, were examined through analysis of cell proliferation and protein expression. Results: The level of phosphorylated AKT (p-AKT) was increased in the primary melanoma cells after treatment with vemurafenib, and the basal level of p-AKT was increased in vemurafenib-resistant melanoma cells. Notably, resveratrol both alone and in combination with vemurafenib effectively suppressed cell proliferation and AKT phosphorylation in both parental and vemurafenib-resistant melanoma cells. Conclusion: Vemurafenib resistance can be reversed by addition of resveratrol in patients undergoing treatment with BRAF inhibitors.

Monday, June 27, 2016 12:52 PM|TANG, T., ELDABAJE, R., YANG, L.|Anticancer Research recent issues|Labels: BRAF, melanoma, skin cancer

Compared to early-stage melanoma when surgical excision is possible, metastatic disease continues to offer a much grimmer prognosis as traditional chemotherapy treatment regimens offer relatively little survival benefit. This has led to changes in treatment approaches over the preceding two decades as contemporary methods for the treatment of advanced or metastatic melanoma now involve a number of biological modalities, which include immunotherapeutic approaches, targeted therapies and epigenetic modification therapies. Clinically available immunotherapeutic agents include interleukin 2 (IL-2), as well as drugs targeting the important immune checkpoint molecules, such as cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1). The targeted therapeutic agents modulate specific pro-oncogenic mutations such as v-Raf murine sarcoma viral oncogene homolog B (BRAF), receptor tyrosine kinases, MEK inhibitors and potential future therapeutic targets, such as the CDK4/CDK6, PTEN and GNAQ/GNA11 genes. Additionally, an increasing understanding of the role of epigenetic alterations in the development and progression of melanoma now offers a new potential drug target. Several of these agents have shown promising results; however, in many investigations, combinations of different therapeutic approaches, each with different mechanisms of action, have yielded improved outcomes as treatment regimens continue to be further optimized by active research and patient disease sub-group analyses. This review summarizes the novel biological agents and new treatments, directly contributing to the significant improvement of biological therapies and markedly advancing knowledge of clinical application of newly approved and developed therapies in treatment of patients with metastatic melanoma.

Saturday, June 18, 2016 2:05 PM|Benjamin Besse; Harry J M Groen; Pierre-;Jean Souquet; Elisabeth Quoix; Christina S Baik; Fabrice Barlesi; Tae Min Kim; Julien Mazieres; Silvia Novello; James R Rigas; Allison Upalawanna; Anthony M D'Amelio; Pingkuan Zhang; Bijoyesh Mookerjee; Bruce E Johnson|JournalTOCs API - The Lancet Oncology (50 articles)|Labels: BRAF, MapK, lung cancer, skin cancer, clinical trial

Dabrafenib plus trametinib in patients with previously treated BRAFV600E-mutant metastatic non-small cell lung cancer: an open-label, multicentre phase 2 trial
David Planchard Benjamin Besse; Harry J M Groen; Pierre-Jean Souquet; Elisabeth Quoix; Christina S Baik; Fabrice Barlesi; Tae Min Kim; Julien Mazieres; Silvia Novello; James R Rigas; Allison Upalawanna; Anthony M D'Amelio; Pingkuan Zhang; Bijoyesh Mookerjee; Bruce E Johnson
The Lancet Oncology, Vol. 17, No. 7 (2016) pp. 984 - 993
Publication date: Available online 6 June 2016 Source:The Lancet Oncology Author(s): David Planchard, Benjamin Besse, Harry J M Groen, Pierre-Jean Souquet, Elisabeth Quoix, Christina S Baik, Fabrice Barlesi, Tae Min Kim, Julien Mazieres, Silvia Novello, James R Rigas, Allison Upalawanna, Anthony M D'Amelio, Pingkuan Zhang, Bijoyesh Mookerjee, Bruce E Johnson Background BRAF mutations act as an oncogenic driver via the mitogen-activated protein kinase (MAPK) pathway in non-small cell lung cancer (NSCLC). BRAF inhibition has shown antitumour activity in patients with BRAF V600E-mutant NSCLC. Dual MAPK pathway inhibition with BRAF and MEK inhibitors in BRAF V600E-mutant NSCLC might improve efficacy over BRAF inhibitor monotherapy based on observations in BRAF V600-mutant melanoma. We aimed to assess the antitumour activity and safety of dabrafenib plus trametinib in patients with BRAF V600E-mutant NSCLC. Methods In this phase 2, multicentre, non-randomised, open-label study, we enrolled adult patients (aged ≥18 years) with pretreated metastatic stage IV BRAF V600E-mutant NSCLC who had documented tumour progression after at least one previous platinum-based chemotherapy and had had no more than three previous systemic anticancer therapies. Patients with previous BRAF or MEK inhibitor treatment were ineligible. Patients with brain metastases were allowed to enrol only if the lesions were asymptomatic, untreated (or stable more than 3 weeks after local therapy if treated), and measured less than 1 cm. Enrolled patients received oral dabrafenib (150 mg twice daily) plus oral trametinib (2 mg once daily) in continuous 21-day cycles until disease progression, unacceptable adverse events, withdrawal of consent, or death. The primary endpoint was investigator-assessed overall response, which was assessed by intention to treat in the protocol-defined population (patients who received second-line or later treatment); safety was also assessed in this population and was assessed at least once every 3 weeks, with adverse events, laboratory values, and vital signs graded according to the Common Terminology Criteria for Adverse Events version 4.0. The study is ongoing but no longer recruiting patients. This trial is registered with ClinicalTrials.gov, number NCT01336634. Findings Between Dec 20, 2013, and Jan 14, 2015, 59 patients from 30 centres in nine countries across North America, Europe, and Asia met eligibility criteria. Two patients who had previously been untreated due to protocol deviation were excluded; thus, 57 eligible patients were enrolled. 36 patients (63·2% [95% CI 49·3–75·6]) achieved an investigator-assessed overall response. Serious adverse events were reported in 32 (56%) of 57 patients and included pyrexia in nine (16%), anaemia in three (5%), confusional state in two (4%), decreased appetite in two (4%), haemoptysis in two (4%), hypercalcaemia in two (4%), nausea in two (4%), and cutaneous squamous cell carcinoma in two (4%). The most common grade 3–4 adverse events were neutropenia in five patients (9%), hyponatraemia in four (7%), and anaemia in three (5%). Four patients died during the study from fatal adverse events judged to be unrelated to treatment (one retroperitoneal haemorrhage, one subarachnoid haemorrhage, one respiratory distress, and one from disease progression that was more severe than typical progression, as assessed by the investigator). Interpretation Dabrafenib plus trametinib could represent a new targeted therapy with robust antitumour activity and a manageable safety profile in patients with BRAF V600E-mutant NSCLC. Funding GlaxoSmithKline.

Saturday, June 18, 2016 2:05 PM|Paul Mitchell|JournalTOCs API - The Lancet Oncology (50 articles)|Labels: BRAF, lung cancer

Combined BRAF and MEK inhibition in BRAF-mutant NSCLC
Gareth Rivalland Paul Mitchell
The Lancet Oncology, Vol. 17, No. 7 (2016) pp. 860 - 862
Publication date: Available online 6 June 2016 Source:The Lancet Oncology Author(s): Gareth Rivalland, Paul Mitchell

Wednesday, June 15, 2016 12:56 PM|Niessner, H., Schmitz, J., Tabatabai, G., Schmid, A., Calaminus, C., Sinnberg, T., Weide, B., Eigentler, T. K., Garbe, C., Schittek, B., Quintanilla-Fend, L., Bender, B., Mai, M., Praetorius, C., Beissert, S., Schackert, G., Muders, M., Meinhardt, M., Baretton, G. B., Dummer, R., Flaherty, K. T., Pichler, B. J., Kulms, D., Westphal, D., Meier, F.|Clinical Cancer Research Online First Articles|Labels: AKT, BRAF, melanoma, skin cancer

Purpose: Great advances have recently been made in treating patients with metastatic melanoma. However, existing therapies are less effective on cerebral than extracerebral metastases. This highlights the potential role of the brain environment on tumor progression and drug resistance and underlines the need for "brain-specific" therapies. We previously showed that the PI3K-AKT survival pathway is hyperactivated in brain but not extracerebral melanoma metastases and that astrocyte-conditioned medium activates AKT in melanoma cells in vitro. We therefore tested the PI3K inhibitor buparlisib as an antitumor agent for melanoma brain metastases. Experimental Design and Results: Buparlisib inhibited AKT activity, decreased proliferation and induced apoptosis in metastatic melanoma cell lines and short-term brain melanoma cells, irrespective of their BRAF and NRAS mutation status. Additionally, buparlisib inhibited hyperactivated AKT and induced apoptosis in melanoma cells that were stimulated with astrocyte-conditioned medium. The growth of tumors induced by injecting human BRAF- and NRAS-mutant metastatic melanoma cells into the brain of mice was significantly inhibited by buparlisib. Conclusions: These results emphasize the value of targeting the PI3K pathway as a strategy to develop drugs for melanoma brain metastases.

Tuesday, March 22, 2016 12:51 PM|Pramio, D. T., Pennacchi, P. C., Maria-Engler, S. S., Campos, A. H. J. F. M., Duprat, J. P., Carraro, D. M., Krepischi, A. C. V.|Journal of Investigative Medicine recent issues|Labels: BRAF, telomerase, melanoma, skin cancer

Epigenetic dysregulation is an important emerging hallmark of cutaneous melanoma development. The global loss of DNA methylation in gene-poor regions and transposable DNA elements of cancer cells contributes to increased genomic instability. Long interspersed element-1 (LINE-1) sequences are the most abundant repetitive sequence of the genome and can be evaluated as a surrogate marker of the global level of DNA methylation. In this work, LINE-1 methylation levels were evaluated in cutaneous melanomas and normal melanocyte primary cell cultures to investigate their possible association with both distinct clinicopathological characteristics and tumor mutational profile. A set of driver mutations frequently identified in cutaneous melanoma was assessed by sequencing (actionable mutations in BRAF, NRAS, and KIT genes, and mutations affecting the TERT promoter) or multiplex ligation-dependent probe amplification (MLPA) (CDKN2A deletions). Pyrosequencing was performed to investigate the methylation level of LINE-1 and CDKN2A promoter sequences. The qualitative analysis showed a trend toward an association between LINE-1 hypomethylation and CDKN2A inactivation (p=0.05). In a quantitative approach, primary tumors, mainly the thicker ones (>4 mm), exhibited a trend toward LINE-1 hypomethylation when compared with control melanocytes. To date, this is the first study reporting in cutaneous melanomas a possible link between the dysregulation of LINE-1 methylation and the presence of driver mutations.

Monday, March 14, 2016 7:00 PM|Proceedings of the National Academy of Sciences|MedWorm: Non-Small Cell Lung Cancer|Comments|Labels: BRAF, EGFR, FAK, lung cancer
We read with great interest the article by Davare et al. (1) on structural insight of ROS1 tyrosine kinase inhibitors. Non-small cell lung cancer (NSCLC) is no longer a single disease but a collection of genetically heterogeneous tumors with different therapeutic options [e.g., aberrations in EGFR, BRAF, HER2/Neu, RET, anaplastic... (Source: Proceedings of the National Academy of Sciences)
Thursday, March 10, 2016 6:00 PM|Cancer|Cancer via MedWorm.com|Comments|Labels: BRAF, RAS, thymic
CONCLUSIONSThe coexistence of BRAF or RAS mutations enhanced the prognostic effects of TERT promoter mutations. Furthermore, TERT promoter mutations strengthened the predictions of mortality and recurrence by the ATA and TNM staging systems, particularly for high‐risk patients with DTC. Cancer 2016. © 2016 American Cancer Society. (Source: Cancer)
Tuesday, March 8, 2016 9:36 AM|Pandzic, T., Larsson, J., He, L., Kundu, S., Ban, K., Akhtar-Ali, M., Hellström, A. R., Schuh, A., Clifford, R., Blakemore, S. J., Strefford, J. C., Bauman, T. S., Lopez-Guillermo, A., Campo, E., Ljungström, V., Mansouri, L., Rosenquist, R., Sjöblom, T., Hellström, M.|Clinical Cancer Research Online First Articles|Labels: BRAF, MapK, CLL

Purpose: To identify resistance mechanisms for the chemotherapeutic drug fludarabine in chronic lymphocytic leukemia (CLL), as innate and acquired resistance to fludarabine-based chemotherapy represents a major challenge for long-term disease control. Experimental Design: We employed piggyBac transposon-mediated mutagenesis, combined with next-generation sequencing, to identify genes that confer resistance to fludarabine in a human CLL cell line. Results: In total, this screen identified 782 genes with transposon integrations in fludarabine-resistant pools of cells. One of the identified genes is a known resistance mediator DCK (deoxycytidine kinase), which encodes an enzyme that is essential for the phosphorylation of the pro-drug to the active metabolite. BMP2K, a gene not previously linked to CLL, was also identified as a modulator of response to fludarabine. In addition, ten out of 782 transposon-targeted genes had previously been implicated in treatment resistance based on somatic mutations seen in patients refractory to fludarabine-based therapy. Functional characterization of these genes supported a significant role for ARID5B and BRAF in fludarabine sensitivity. Finally, pathway analysis of transposon-targeted genes and RNA-seq profiling of fludarabine-resistant cells suggested deregulated MAPK signaling as involved in mediating drug resistance in CLL. Conclusions: To our knowledge, this is the first forward genetic screen for chemotherapy resistance in CLL. The screen pinpointed novel genes and pathways involved in fludarabine resistance along with previously known resistance mechanisms. Transposon screens can therefore aid interpretation of cancer genome sequencing data in the identification of genes modifying sensitivity to chemotherapy

Sunday, February 28, 2016 6:00 PM|EMBO Journal|MedWorm: Cancer Therapy|Comments|Labels: BRAF, melanoma, skin cancer
The activation of transcriptional coactivators YAP and its paralog TAZ has been shown to promote resistance to anti-cancer therapies. YAP/TAZ activity is tightly coupled to actin cytoskeleton architecture. However, the influence of actin remodeling on cancer drug resistance remains largely unexplored. Here, we report a pivotal role of actin remodeling in YAP/TAZ-dependent BRAF inhibitor resistance in BRAF V600E mutant melanoma cells. Melanoma cells resistant to the BRAF inhibitor PLX4032 exhibit an increase in actin stress fiber formation, which appears to promote the nuclear accumulation of YAP/TAZ. Knockdown of YAP/TAZ reduces the viability of resistant melanoma cells, whereas overexpression of constitutively active YAP induces resistance. Moreover, inhibition of actin polymerization and...
Thursday, February 25, 2016 6:00 PM|Critical Reviews in Oncology Hematology|MedWorm: Non-Small Cell Lung Cancer|Comments|Labels: ALK, BRAF, EGFR, lung cancer
The low survival rates observed in Non-Small cell lung cancer (NSCLC) patients occurs due to a dangerous association of late detection and limited efficacy of the available treatments (Jemal et al., 2009). The discovery of common oncogene drivers, such as epidermal growth factor receptor (EGFR) mutations, anaplastic lymphoma kinase fusion (EML-ALK) and proto-oncogene tyrosine-protein kinase ROS1 rearrangements, has led to the development of new accurate and efficient targeted therapies, radically improving the clinical outcomes of patients that harbor these mutations, opening the new era of targeted therapy in lung cancer (Solomon et al., 2014; Rosell et al., 2012). (Source: Critical Reviews in Oncology Hematology)
Wednesday, February 17, 2016 6:00 PM|Cancer Research|MedWorm: Non-Small Cell Lung Cancer|Comments|Labels: BRAF, EGFR, breast cancer, skin cancer, clinical trial
Background: Nivo is an inhibitory antibody against programmed death receptor-1 (PD-1), a regulator of antitumor immunity. Nivo is approved for treatment of unresectable or metastatic melanoma and disease progression (PD) following ipilimumab or, in BRAF V600 mutation–positive melanoma, following a BRAF inhibitor, and for metastatic squamous non-small cell lung cancer (NSCLC) following PD during or after platinum-based chemotherapy. Nivo and other immune checkpoint inhibitors are also being investigated in other tumor-types. nab-P is a novel taxane formulation and does not require prophylaxis with immunosuppressive steroids. It has demonstrated superior efficacy over control regimens in phase III studies of MBC, pancreatic cancer, and NSCLC. This open-label, 6-arm, multicenter phase I tri...
Wednesday, February 17, 2016 11:44 AM|Expert Opinion on Emerging Drugs|MedWorm: Non-Small Cell Lung Cancer|Comments|Labels: ALK, BRAF, EGFR, lung cancer
This article reviews the next-generation kinase inhibitors targeting EGFR and ALK-positive NSCLC. In addition, targeted kinase inhibitors in clinical development for other specific molecular subtypes of NSCLC are covered, including ROS1, BRAF, RET, HER2, KRAS (upstream of the MEK kinase), MET, PIK3CA, FGFR1, DDR2, VEGFR and AAK. EXPERT OPINION: In EGFR-mutant NSCLC, there are several kinase inhibitors with promising activity, most notably dacomitinib and CO-1686 in tumors with acquired resistance to EGFR-targeted therapy. Next-generation ALK inhibitors appear to have greater potency than crizotinib and several ongoing trials may shed light on their role in both ALK- and ROS1-positive NSCLC. While there is optimism regarding the role of kinase inhibitors in other molecular subtypes, the...
Wednesday, February 17, 2016 9:36 AM|Expert Review of Anticancer Therapy|MedWorm: Non-Small Cell Lung Cancer|Comments|Labels: ALK, BRAF, EGFR, fibrosarcoma, lung cancer
Authors: Khanal N, Ganti AK Abstract Lung cancer is the leading cause of cancer-related deaths in United States, accounting for more than one-fourth of the deaths annually. Although comparatively rare and relatively less studied, genetic abnormalities other than epidermal growth factor receptor (EGFR) mutations, anaplastic lymphoma kinase (ALK) rearrangements, and Kirsten rat sarcoma (KRAS) mutations account for significant proportion of the driver mutations identified thus far. The targeted agents against B-rapidly accelerated fibrosarcoma (BRAF) V600E mutation, MNNG-HOS transforming gene (MET) pathway, ROS1 rearrangement, rearranged during transfection (RET) rearrangement, and HER2 pathways offer promising therapeutic options. Recruiting patients with these rarer mutations to wel...
Tuesday, February 2, 2016 6:00 PM|Cancer Chemotherapy and Pharmacology|MedWorm: Non-Small Cell Lung Cancer|Comments|Labels: ALK, BRAF, EGFR, lung cancer, biomarker diagnostic
Conclusion Combined analysis of these commonly studied genes may promote the individual treatment in NSCLC. RRM1 may be a prognostic and predictive biomarker for PFS in patients with NSCLC who received platinum-based adjuvant chemotherapy, and combining EGFR mutation and RRM1 expression or combining ERCC1 and RRM1 expression can enhance prognostic and predictive power for PFS. (Source: Cancer Chemotherapy and Pharmacology)
Monday, February 1, 2016 1:00 AM|Buchbinder, Elizabeth I.; Desai, Anupam|American Journal of Clinical Oncology - Most Popular Articles|Labels: BRAF, skin cancer
imageThe cytotoxic T-lymphocyte–associated antigen 4 (CTLA-4) and programmed death 1 (PD-1) immune checkpoints are negative regulators of T-cell immune function. Inhibition of these targets, resulting in increased activation of the immune system, has led to new immunotherapies for melanoma, non–small cell lung cancer, and other cancers. Ipilimumab, an inhibitor of CTLA-4, is approved for the treatment of advanced or unresectable melanoma. Nivolumab and pembrolizumab, both PD-1 inhibitors, are approved to treat patients with advanced or metastatic melanoma and patients with metastatic, refractory non-small cell lung cancer. In addition the combination of ipilimumab and nivolumab has been approved in patients with BRAF WT metastatic or unresectable melanoma. The roles of CTLA-4 and PD-1 in inhibiting immune responses, including antitumor responses, are largely distinct. CTLA-4 is thought to regulate T-cell proliferation early in an immune response, primarily in lymph nodes, whereas PD-1 suppresses T cells later in an immune response, primarily in peripheral tissues. The clinical profiles of immuno-oncology agents inhibiting these 2 checkpoints may vary based on their mechanistic differences. This article provides an overview of the CTLA-4 and PD-1 pathways and implications of their inhibition in cancer therapy.
Saturday, January 23, 2016 6:00 PM|Journal of Genetics and Genomics|MedWorm: Cancer Therapy|Comments|Labels: ALK, BRAF, EGFR, lung cancer
Publication date: Available online 15 September 2015 Source:Journal of Genetics and Genomics Author(s): Jingwu Xie, Xiaoli Zhang There is high expectation for significant improvements in cancer patient care after completion of the human genome project in 2003. Through pains-taking analyses of genomic profiles in cancer patients, a number of targetable gene alterations have been discovered, with some leading to novel therapies, such as activating mutations of EGFR, BRAF and ALK gene fusions. As a result, clinical management of cancer through targeted therapy has finally become a reality for a subset of cancers, such as lung adenocarcinomas and melanomas. In this review, we summarize how gene mutation discovery leads to new treatment strategies using non-small cell lung cancer (NSCLC) as...
Friday, January 22, 2016 4:15 PM|News-Medical.Net Cetuximab News Feed|Comments|Labels: BRAF, 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.
Thursday, January 14, 2016 7:00 PM|April Salama|Cancers|Labels: BRAF, melanoma, skin cancer
Cutaneous melanoma is one of the most aggressive forms of skin cancer, and is correlated with a large proportion of skin cancer-related deaths. Therapy for cutaneous melanoma has advanced greatly through careful identification of therapeutic targets and the development of novel immunotherapeutic approaches. The identification of BRAF as well as other driver mutations, have allowed for a specialized approach to treatment. In addition, immune checkpoint inhibition has dramatically changed the treatment landscape over the past 5–10 years. The successful targeting of CTLA-4, as well as PD-1/PD-L1, has been translated into meaningful clinical benefit for patients, with multiple other potential agents in development. Systemic therapy for cutaneous melanoma is becoming more nuanced and often takes a multifaceted strategy. This review aims to discuss the benefits and limitations of current therapies in systemic melanoma treatment as well as areas of future development.
Thursday, January 14, 2016 6:00 PM|The Lancet|MedWorm: Non-Small Cell Lung Cancer|Comments|Labels: ALK, BRAF, EGFR, lung cancer
This study included patients with advanced NSCLC, who were routinely screened for EGFR mutations, ALK rearrangements, as well as HER2 (ERBB2), KRAS, BRAF, and PIK3CA mutations by 28 certified regional genetics centres in France. Patients were assessed consecutively during a 1-year period from April, 2012, to April, 2013. We measured the frequency of molecular alterations in the six routinely screened genes, the turnaround time in obtaining molecular results, and patients' clinical outcomes. This study is registered with ClinicalTrials.gov, number NCT01700582. Findings 18 679 molecular analyses of 17 664 patients with NSCLC were done (of patients with known data, median age was 64·5 years [range 18–98], 65% were men, 81% were smokers or former smokers, and 76% had adenocarcinoma). T...
Wednesday, January 6, 2016 6:00 PM|Molecular Cancer Therapeutics|MedWorm: Non-Small Cell Lung Cancer|Comments|Labels: BRAF, EGFR, clinical trial
Conclusions: To date, treatment with RXDX-105 demonstrates an acceptable safety profile. Improvements in bioavailability are being investigated. Dose escalation continues.Citation Format: Ding Wang, Manish Patel, Marwan Fakih, A. Craig Lockhart, Anthony J. Olszanski, Rupal Patel, Peter D. Brown, Jennifer W. Oliver, Pratik S. Multani. A phase 1 study of RXDX-105, an oral RET, BRAF and EGFR tyrosine kinase inhibitor, in patients with advanced or metastatic cancers. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C188. (Source: Molecular Cancer Therapeutics)
Wednesday, January 6, 2016 6:00 PM|Molecular Cancer Therapeutics|MedWorm: Cancer Therapy|Comments|Labels: BRAF, melanoma, skin cancer
ConclusionOur results implicate the actin cytoskeleton in the induction of YAP/TAZ-dependent resistance to vemurafenib, and inhibition of actin remodeling might be a promising synthetic lethal strategy to suppress resistance in BRAF inhibitor therapies.Citation Format: Min Hwan Kim, Joon Kim. Actin remodeling confers BRAF inhibitor resistance to melanoma cells through YAP/TAZ activation. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr B50. (Source: Molecular Cancer Therapeutics)

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Friday, January 1, 2016 6:00 PM|Journal of Molecular Diagnostics|MedWorm: Non-Small Cell Lung Cancer|Comments|Labels: BRAF, EGFR, lung cancer
Personalized medicine has gained increasing importance in clinical oncology, and several clinically important biomarkers are implemented in routine practice. In an effort to guarantee high quality of molecular testing in France, three subsequent external quality assessment rounds were organized at the initiative of the National Cancer Institute between 2012 and 2014. The schemes included clinically relevant biomarkers for metastatic colorectal (KRAS, NRAS, BRAF, PIK3CA, microsatellite instability) and non-small cell lung cancer (EGFR, KRAS, BRAF, PIK3CA, ERBB2), and they represent the first multigene/multicancer studies throughout Europe. (Source: Journal of Molecular Diagnostics)

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