Oncology Intelligence

FAK, a cytosolic kinase, is involved in cellular adhesion, spreading processes, cell locomotion, mitogen response, and cell survival. FAK activity elicits intracellular signal transduction pathways that promote the turn-over of cell contacts with the ECM, promoting cell migration. Throughout apoptosis, FAK is an important contributor to cell rounding, loss of focal contacts and apoptotic membrane formations such as blebbing. Anoikis is a form of programmed cell death which is induced by anchorage-dependent cells detaching from the surrounding ECM.(1, 2) Most cells express FAK and FAK is required during development.(3) FAK is typically located at structures known as focal adhesion, these are multi-protein structures that link the ECM to the cytoplasmic cytoskeleton. When dynamic microtubules make contact, focal adhesions disassemble and FAK plays a role in this process. Phosphorylation of tyrosine 925 and the subsequent recruitment of Grb2 and dynamin into a complex with FAK are required for microtubule-induced disassembly of focal adhesions. There is a the link between FAK and the regulation of microtubule structure and organization.(4, 5) Additional components of focal adhesions include actin, filamin, vinculin, talin, paxillin, tensin, and RSU-1.(6)
FAK has four domains. Two of these domains, the N-terminal FERM domain and the kinase domain form an auto-inhibitory interaction. This interaction, the result of hydrophobic interactions, prevents the activation of the kinase domain, thereby preventing the signalling function of FAK.(6) FAK-dependent kinase activity is performed mostly by the SFKs, Src or Fyn, which are recruited and activated by a dentate interaction between their SH2 and SH3 domains and FAK's p-Tyr397 and PR1 motifs.(7) FAK is phosphorylated in response to INT engagement, growth factor stimulation, and the action of mitogenic neuropeptides.(6) FAK also binds to PI-3k, Grb-7, EGFR, VEGFR, p53, Jak2, TGFB1I1, STAT1, PTEN, and MAPK8IP3.(3, 6, 8-14).
During early apoptotic signaling in human endothelial cells, FAK is cleaved by caspase 3 at Asp-772, generating two FAK fragments. The smaller FAK fragment, killer FAT, and becomes the domain associated with death signaling. Throughout apoptosis, FAK is an important contributor to cell rounding, loss of focal contacts, and apoptotic membrane formations such as blebbing.(9) Overexpression of FAK leads to inhibition of apoptosis and an increase in the prevalence of metastatic tumors.(6, 9)
FAK may play a role in the regulation of the tumor suppressor p53. Overexpression of FAK leads to inhibition of apoptosis and an increase in the prevalence of metastatic tumors.(3, 9) It has been shown that when FAK was blocked, BC cells became less metastatic due to decreased mobility.(3) Increased FAK expression is found in several different cancers including laryngeal, esophageal, CRC, BC, PC, and melanoma.(15)
Existing FAK-based therapeutics focus on inhibiting the kinase's catalytic function.(16) The major downstream signaling pathway activated by HER2 cross-talk is PI3K/mTOR, and a potential integrator of receptor crosstalk is Src-FAK signaling. PI3K, Src, and FAK have independently been implicated in trastuzumab resistance.(17) TGF-mediated trastuzumab resistance was overcome by Src-FAK inhibition.(17, 18) Targeting the FAK scaffold is also a feasible and promising approach for developing highly specific therapeutics that disrupt FAK signaling pathways in cancer.(16)



1. Inbal B, Bialik S, Sabanay I, Shani G, Kimchi A. DAP kinase and DRP-1 mediate membrane blebbing and the formation of autophagic vesicles during programmed cell death. The Journal of cell biology. 2002;157(3):455-68.

2. Frisch SM, Schaller M, Cieply B. Mechanisms that link the oncogenic epithelial-mesenchymal transition to suppression of anoikis. Journal of Cell Science. 2013;126(1):21-9.

3. PTK2. [cited]; Available from:

4. Schaller MD. Cellular functions of FAK kinases: insight into molecular mechanisms and novel functions. Journal of Cell Science. 2010;123(7):1007-13.

5. Palazzo AF, Eng CH, Schlaepfer DD, Marcantonio EE, Gundersen GG. Localized stabilization of microtubules by integrin-and FAK-facilitated Rho signaling. Science. 2004;303(5659):836-9.

6. Parsons MJ GD. Mitochondria in cell death. . Essays Biochem. 2010;47:99-114. PMCID: 20533903.

7. Arold ST. How focal adhesion kinase achieves regulation by linking ligand binding, localization and action. Current opinion in structural biology. 2011;21(6):808-13.

8. Guinebault C, Payrastre B, Racaud-Sultan C, Mazarguil H, Breton M, Mauco G, et al. Integrin-dependent translocation of phosphoinositide 3-kinase to the cytoskeleton of thrombin-activated platelets involves specific interactions of p85 alpha with actin filaments and focal adhesion kinase. The Journal of cell biology. 1995;129(3):831-42.

9. Mehlen P, Puisieux A. Metastasis: a question of life or death. Nature Reviews Cancer. 2006;6(6):449-58.

10. Dunn KB, Heffler M, Golubovskaya V. Evolving therapies and FAK inhibitors for the treatment of cancer. Anti-cancer agents in medicinal chemistry. 2010;10(10):722.

11. Golubovskaya VM, Cance WG. Focal adhesion kinase and p53 signaling in cancer cells. International review of cytology. 2007;263:103-53.

12. Zhu T, Goh EL, Lobie PE. Growth hormone stimulates the tyrosine phosphorylation and association of p125 focal adhesion kinase (FAK) with JAK2 Fak is not required for stat-mediated transcription. Journal of Biological Chemistry. 1998;273(17):10682-9.

13. Ryu H, Lee J-H, Kim KS, Jeong S-M, Kim P-H, Chung H-T. Regulation of neutrophil adhesion by pituitary growth hormone accompanies tyrosine phosphorylation of Jak2, p125FAK, and paxillin. The Journal of Immunology. 2000;165(4):2116-23.

14. Matsuya M, Sasaki H, Aoto H, Mitaka T, Nagura K, Ohba T, et al. Cell adhesion kinase ? forms a complex with a new member, Hic-5, of proteins localized at focal adhesions. Journal of Biological Chemistry. 1998;273(2):1003-14.

15. Infusino GA, Jacobson JR. Endothelial FAK as a therapeutic target in disease. Microvascular research. 2012;83(1):89-96.

16. Cance WG, Kurenova E, Marlowe T, Golubovskaya V. Disrupting the Scaffold to Improve Focal Adhesion Kinase-Targeted Cancer Therapeutics. Science signaling. 2013;6(268):pe10.

17. Nahta R. Pharmacological strategies to overcome HER2 cross-talk and Trastuzumab resistance. Current medicinal chemistry. 2012;19(7):1065.

18. Wang SE, Xiang B, Zent R, Quaranta V, Pozzi A, Arteaga CL. Transforming growth factor ? induces clustering of HER2 and integrins by activating Src-focal adhesion kinase and receptor association to the cytoskeleton. CANCER RESEARCH. 2009;69(2):475-82.

Sunday, October 2, 2016 10:05 PM|Gainor, J. F., Dardaei, L., Yoda, S., Friboulet, L., Leshchiner, I., Katayama, R., Dagogo-Jack, I., Gadgeel, S., Schultz, K., Singh, M., Chin, E., Parks, M., Lee, D., DiCecca, R. H., Lockerman, E., Huynh, T., Logan, J., Ritterhouse, L. L., Le, L. P., Muniappan, A., Digumarthy, S., Channick, C., Keyes, C., Getz, G., Dias-Santagata, D., Heist, R. S., Lennerz, J., Sequist, L. V., Benes, C. H., Iafrate, A. J., Mino-Kenudson, M., Engelman, J. A., Shaw, A. T.|Cancer Discovery current issue|Labels: ALK, FAK, lung cancer

Advanced, anaplastic lymphoma kinase (ALK)–positive lung cancer is currently treated with the first-generation ALK inhibitor crizotinib followed by more potent, second-generation ALK inhibitors (e.g., ceritinib and alectinib) upon progression. Second-generation inhibitors are generally effective even in the absence of crizotinib-resistant ALK mutations, likely reflecting incomplete inhibition of ALK by crizotinib in many cases. Herein, we analyzed 103 repeat biopsies from ALK-positive patients progressing on various ALK inhibitors. We find that each ALK inhibitor is associated with a distinct spectrum of ALK resistance mutations and that the frequency of one mutation, ALKG1202R, increases significantly after treatment with second-generation agents. To investigate strategies to overcome resistance to second-generation ALK inhibitors, we examine the activity of the third-generation ALK inhibitor lorlatinib in a series of ceritinib-resistant, patient-derived cell lines, and observe that the presence of ALK resistance mutations is highly predictive for sensitivity to lorlatinib, whereas those cell lines without ALK mutations are resistant.

Significance: Secondary ALK mutations are a common resistance mechanism to second-generation ALK inhibitors and predict for sensitivity to the third-generation ALK inhibitor lorlatinib. These findings highlight the importance of repeat biopsies and genotyping following disease progression on targeted therapies, particularly second-generation ALK inhibitors. Cancer Discov; 6(10); 1118–33. ©2016 AACR.

See related commentary by Qiao and Lovly, p. 1084.

This article is highlighted in the In This Issue feature, p. 1069

Sunday, October 2, 2016 10:05 PM|Zhou, Y., Dang, J., Chang, K.-Y., Yau, E., Aza-Blanc, P., Moscat, J., Rana, T. M.|Cancer Research recent issues|Labels: FAK, RAS
Global miRNA functional screens can offer a strategy to identify synthetic lethal interactions in cancer cells that might be exploited therapeutically. In this study, we applied this strategy to identify novel gene interactions in KRAS-mutant cancer cells. In this manner, we discovered miR-1298, a novel miRNA that inhibited the growth of KRAS-driven cells both in vitro and in vivo. Using miR-TRAP affinity purification technology, we identified the tyrosine kinase FAK and the laminin subunit LAMB3 as functional targets of miR-1298. Silencing of FAK or LAMB3 recapitulated the synthetic lethal effects of miR-1298 expression in KRAS-driven cancer cells, whereas coexpression of both proteins was critical to rescue miR-1298–induced cell death. Expression of LAMB3 but not FAK was upregulated by mutant KRAS. In clinical specimens, elevated LAMB3 expression correlated with poorer survival in lung cancer patients with an oncogenic KRAS gene signature, suggesting a novel candidate biomarker in this disease setting. Our results define a novel regulatory pathway in KRAS-driven cancers, which offers a potential therapeutic target for their eradication. Cancer Res; 76(19); 5777–87. ©2016 AACR.
Thursday, September 29, 2016 6:00 PM|Cancer Research UK|Cancer Research UK News|Labels: FAK

Science blog

In a corner of our head office, sits a team of six people. And since 2010, they’ve been quietly, but resolutely helping to develop new treatment options for cancer patients.

This is the team behind the Combinations Alliance, one of several projects run through the Experimental Cancer Medicine Centre network (ECMC).

The Alliance brings together UK researchers and drug companies from around the world to explore new combinations of cancer drugs. By combining multiple drugs in a single clinical trial, they can test whether or not the combination is better at treating cancer than the standard treatment available.

It’s a unique scheme that’s increasing treatment options for patients, and tackling drug resistance – arguably one of the biggest problems in cancer treatment.

And a new deal signed today will see a combination of drugs tested in mesothelioma, non-small cell lung and pancreatic cancers for the first time.

A match made in heaven

The Combinations Alliance works very much like a match-making agency for researchers and drug companies. The goal of developing these relationships is to hopefully launch clinical trials that test promising new combinations of drugs to treat different types of cancer.

The Combinations Alliance focuses on therapies that wouldn’t progress without our support

– Dr Ian Walker, director of clinical research and strategic partnerships at Cancer Research UK

So why exactly was the Combinations Alliance set up? Surely companies have thought of collaborating in this way before? Well, this can be tricky. Each company has a different way of working.

So it’s often much simpler for many drug companies to go it alone. But they’re beginning to see how limiting this approach is. Despite huge resources, it’s simply impossible for any company to test every combination of drugs.

But our Combinations Alliance offers a structured way for drug companies – alongside researchers – to work together. And, ultimately, get better treatments to patients, sooner.

How the initiative works

Our team begins by meeting a drug company that’s developing promising cancer drugs. Researchers within our ECMC network then consider which drugs could work together in a clinical trial.

So far twelve partners have signed onto the scheme, with many more set to join in the next few years.

Alternatively, the team receives an idea from a researcher to combine two or more drugs, which aren’t necessarily owned by an existing Alliance partner.

The team then approaches the company who owns that drug for permission to use it in a clinical trial.

Most trials to date have tested a single company’s experimental new drug in combination with a drug that’s already available as standard treatment for patients – or with radiotherapy. There have also been a few trials involving two new experimental drugs owned by the same company.

But for the first time today, the Alliance has brought together two drug companies to test an exciting combination of two drugs: one an immunotherapy drug, the other a so-called ‘targeted’ cancer treatment.

And it’s thanks to the inspirational idea of two researchers – Dr Stefan Symeonides, at the University of Edinburgh, and Professor Dean Fennell, at the University of Leicester.

The bright idea

Together, Symeonides and Fennell designed the combination clinical trial that will be managed at Cancer Research UK’s Clinical Trials Unit in Glasgow. It’s an idea that’s based on work from another of our researchers, Professor Margaret Frame, at the University of Edinburgh.

But designing the trial was only one part of the story. To run it, Symeonides and Fennell needed approval to use the drugs, which are owned and made by two different drug companies.

Let’s step back a couple of years to see how the team made this happen.

Finding a drug that works

Back in 2014, a small drug company based just north of New Jersey in Massachusetts in the US, called Verastem, got in touch with the team.

They’re an existing Alliance partner, introduced by Fennell, and have been working on a drug called VS-6063, which switches off a molecule called FAK that’s found inside cells.

The drug works by stopping FAK forming a cellular barrier that blocks the body’s immune system.

But once the barrier is down, how can the body attack the tumour itself?

Symeonides and Fennell believe that Verastem’s drug could work better with another drug that boosts the immune system and the army of cells it unleashes. And they had a good idea of what could work.

The final piece of the jigsaw

Fast-forward to 2015 and the answer to this problem could be found back in New Jersey at a different drug company, MSD, which is one of the largest drug companies in the world.

Although not an Alliance partner at the time, they were in talks with the team, and agreed to let Symeonides and Fennell use an immunotherapy drug called pembrolizumab (Keytruda) for the trial. Pembrolizumab is designed to target an antenna-like molecule that sticks out on the surface of certain forms of immune cell.

Normally, this ‘antenna’ – called the programmed cell death 1 (PD-1) receptor – picks up signals, preventing the immune system from inappropriately reacting to certain triggers. But in people with some types of cancer, the same antenna receives signals stopping the body’s immune system from recognising the cancer cells, allowing the tumour to remain undetected.

Pembrolizumab blocks these signals, jumpstarting the immune system into recognising, targeting and destroying tumours.

Symeonides and Fennell thought that once the barrier surrounding the cancer cells has been taken down by Verastem’s drug, MSD’s pembrolizumab could then activate cancer-killing immune cells to attack the tumour.

Pembrolizumab on its own has shown promise in treating bladder, melanoma, kidney and non-small cell lung cancer, but it’s had little effect in people with other types of cancer. So this would be a chance to see if the drug could treat more types than originally thought.

Fast-forward to today, and both Verastem and MSD have agreed to the use of their drugs in combination for the first time in a clinical trial. The trial will look at whether the two drugs can be used safely together, and test whether the combination is better for treating people with mesothelioma, non-small cell lung and pancreatic cancers – all of which have very low survival.

What’s next?

By working with UK researchers, and drug companies around the world, there are still many more trial ideas to be explored. These aren’t limited to those involving drug combinations, but using radiotherapy and surgery too. It’s just about getting the right people with the right drugs to start working together.

What the Combinations Alliance team have achieved to date is no small feat. They’re responsible for pulling in some of the best science and making sure that these trials are run in the UK (MSD and Verastem are both American companies and logistically, it’d be a lot easier for them to run the trial in the US), so that patients here can benefit first.

It’s early days, the trial isn’t recruiting patients yet – but it should be up and running later this year.

So far 356 people have taken part in a Combinations Alliance trial and have been given another shot at tackling cancer that’s come back.

One trial in particular was so promising that it’s now progressed to the next step, a larger clinical to see how well it actually works in a larger number of people.

The Combinations Alliance team might be sitting quietly in the corner in our office, but make no mistake, they’re causing quite the stir outside it.


Read more
Monday, September 26, 2016 3:00 PM|Jingwei Tian|PLOS ONE Alerts: New Articles|Labels: FAK, gastric

by Wei Zhu, Liang Ye, Jianzhao Zhang, Pengfei Yu, Hongbo Wang, Zuguang Ye, Jingwei Tian

PFKFB3 (6-phosphofructo-2-kinase) synthesizes fructose 2,6-bisphosphate (F2,6P2), which is an allosteric activator of 6-phosphofructo-1-kinase (PFK-1), the rate-limiting enzyme of glycolysis. Overexpression of the PFKFB3 enzyme leads to high glycolytic metabolism, which is required for cancer cells to survive in the harsh tumor microenvironment. The objective of this study was to investigate the antitumor activity of PFK15 (1-(4-pyridinyl)-3-(2-quinolinyl)-2-propen-1-one), a small molecule inhibitor of PFKFB3, against gastric cancer and to explore its potential mechanisms. The effects of PFK15 on proliferation, apoptosis and cell cycle progression in gastric cancer cells were evaluated by cytotoxicity and apoptosis assays, flow cytometry, and western blotting. In addition, the invasion inhibition effects of PFK15 were measured by transwell invasion assay and western blot analysis, and a xenograft tumor model was used to verify the therapeutic effect of PFK15 in vivo. Results showed that PFK15 inhibited the proliferation, caused cell cycle arrest in G0/G1 phase by blocking the Cyclin-CDKs/Rb/E2F signaling pathway, and induced apoptosis through mitochondria in gastric cancer cells. Tumor volume and weight were also significantly reduced upon intraperitoneal injection with PFK15 at 25 mg/kg. In addition, PFK15 inhibited the invasion of gastric cancer cells by downregulating focal adhesion kinase (FAK) expression and upregulating E-cadherin expression. Taken together, our findings indicate that PFK15 is a promising anticancer drug for treating gastric cancer.
Friday, September 23, 2016 10:46 AM|Onclive Articles|Labels: ALK, FAK, lung cancer, clinical trial
Ceritinib improved progression-free survival compared with standard chemotherapy as a first-line treatment for patients with ALK-positive non–small cell lung cancer. 
Friday, September 23, 2016 5:29 AM|Reuters: Healthcare|Labels: FAK, lung cancer
ZURICH, Sept 23 (Reuters) - Novartis's Zykadia drug performed well against a rare form of lung cancer, the Swiss company said on Friday, citing a study it hopes will help it win expanded regulatory approval for the use of the drug.
Friday, September 23, 2016 5:19 AM|Medications/Drugs News Headlines - Yahoo! News|Attachments|Labels: ALK, FAK, lung cancer, clinical trial

A Novartis logo is pictured on its headquarters building in MumbaiBy John Miller ZURICH (Reuters) - Novartis's Zykadia drug performed well against a rare form of lung cancer, the Swiss company said on Friday, citing a study it hopes will help it win expanded regulatory approval for the use of the drug. Novartis released results of a phase III clinical trial of Zykadia, or ceritinib, on previously untreated patients with advanced anaplastic lymphoma kinase-positive (ALK+) non-small cell lung cancer. Patients treated with Zykadia showed a significant improvement in their chances of surviving without the cancer spreading compared with standard chemotherapy, the firm said.

Thursday, September 22, 2016 11:49 PM| Nachrichten zu Pharma|Labels: FAK, lung cancer, clinical trial
BASEL (dpa-AFX) - Novartis (NVS) announced top-line results from its Phase III ASCEND-4 clinical study for Zykadia (ceritinib) in patients with advanced anaplastic lymphoma kinase-positive non-sma...
Thursday, September 22, 2016 11:47 PM| Nachrichten zu Pharma|Labels: ALK, FAK, lung cancer, clinical trial
Novartis International AG / Novartis announces positive top-line results from ASCEND-4, a Phase III trial of Zykadia(R) in untreated adult ALK+ NSCLC patients . Processed and transmitted by Nasdaq...
Thursday, September 15, 2016 7:42 AM|Skinner, H. D., Giri, U., Yang, L., Woo, S. H., Story, M. D., Pickering, C. R., Byers, L. A., Williams, M. D., El-Naggar, A., Wang, J., Diao, L., Shen, L., Fan, Y. H., Molkentine, D. P., Beadle, B. M., Meyn, R. E., Myers, J. N., Heymach, J. V.|Clinical Cancer Research current issue|Labels: FAK, FGFR, HNN

Purpose: Head and neck squamous cell carcinoma (HNSCC) is commonly treated with radiotherapy, and local failure after treatment remains the major cause of disease-related mortality. To date, human papillomavirus (HPV) is the only known clinically validated, targetable biomarkers of response to radiation in HNSCC.

Experimental Design: We performed proteomic and transcriptomic analysis of targetable biomarkers of radioresistance in HPV-negative HNSCC cell lines in vitro, and tested whether pharmacologic blockade of candidate biomarkers sensitized cells to radiotherapy. Candidate biomarkers were then investigated in several independent cohorts of patients with HNSCC.

Results: Increased expression of several targets was associated with radioresistance, including FGFR, ERK1, EGFR, and focal adhesion kinase (FAK), also known as PTK2. Chemical inhibition of PTK2/FAK, but not FGFR, led to significant radiosensitization with increased G2–M arrest and potentiated DNA damage. PTK2/FAK overexpression was associated with gene amplification in HPV-negative HNSCC cell lines and clinical tumors. In two independent cohorts of patients with locally advanced HPV-negative HNSCC, PTK2/FAK amplification was highly associated with poorer disease-free survival (DFS; P = 0.012 and 0.034). PTK2/FAK mRNA expression was also associated with worse DFS (P = 0.03). Moreover, both PTK2/FAK mRNA (P = 0.021) and copy number (P = 0.063) were associated with DFS in the Head and Neck Cancer subgroup of The Cancer Genome Atlas.

Conclusions: Proteomic analysis identified PTK2/FAK overexpression is a biomarker of radioresistance in locally advanced HNSCC, and PTK2/FAK inhibition radiosensitized HNSCC cells. Combinations of PTK2/FAK inhibition with radiotherapy merit further evaluation as a therapeutic strategy for improving local control in HPV-negative HNSCC. Clin Cancer Res; 22(18); 4643–50. ©2016 AACR.

Tuesday, August 30, 2016 6:00 PM|Chi-Wen Luo|International Journal of Molecular Sciences|Labels: FAK, INT, breast cancer
Triple negative breast cancer (TNBC) displays higher risk of recurrence and distant metastasis. Due to absence of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2), TNBC lacks clinically established targeted therapies. Therefore, understanding of the mechanism underlying the aggressive behaviors of TNBC is required for the design of individualized strategies and the elongation of overall survival duration. Here, we supported a positive correlation between β1 integrin and malignant behaviors such as cell migration, invasion, and drug resistance. We found that silencing of β1 integrin inhibited cell migration, invasion, and increased the sensitivity to anti-cancer drug. In contrast, activation of β1 integrin increased cell migration, invasion, and decreased the sensitivity to anti-cancer drug. Furthermore, we found that silencing of β1 integrin abolished Focal adhesion kinese (FAK) mediated cell survival. Overexpression of FAK could restore cisplatin-induced apoptosis in β1 integrin-depleted cells. Consistent to in vitro data, β1 integrin expression was also positively correlated with FAK (p = 0.031) in clinical tissue. More importantly, β1 integrin expression was significantly correlated with patient outcome. In summary, our study indicated that β1 integrin could regulate TNBC cells migration, invasion, drug sensitivity, and be a potential prognostic biomarker in TNBC patient survival.
Sunday, August 14, 2016 10:05 PM|Jen, J., Mansfield, A., Eiken, P. W., Stoddard, S., Pierson, K., Hou, X., Ren, H. Z., Molina, J., Yi, J., Yang, P.|Clinical Cancer Research recent issues|Labels: ALK, EGFR, FAK, lung cancer

Mutations in the EGFR gene or transcript fusion involving EML4-ALK result in oncogenic activation of these oncogenes driving lung adenocarcinoma growth in tumors carrying such an alteration. For lung adenocarcinoma with these genetic changes, small-molecule tyrosine kinase inhibitors have been highly effective at reducing tumor burden and improve the outcome of the patients. Unfortunately, drug resistance eventually develops in these tumors and challenges remain in controlling lung cancer recurrence after targeted therapy.

To overcome recurrence, we initiated a prove-of-principle study to evaluate the feasibility of an integrated strategy utilizing genomic profiles of the tumor and patient-specific tumor xenografts derived (PDX) from biopsies for ex vivo evaluation of antitumor drugs to help guide personalized treatment of lung cancer in patients who have developed resistance to targeted therapy drugs. Our study has three main objectives. 1) Use tumor biopsies obtained at the time of recurrence for oncogene mutation and RNAseq analyses to identify molecular changes in oncogenes and gene pathways that can be potentially targeted. 2) Evaluate drug efficacy and optimize personalized therapy for each patient using PDX models. 3) Assess clinical feasibility and our experience using integrated approaches for lung cancer patients who failed targeted therapy.

This study was approved by the Mayo Clinic Institutional Review Board (IRB) and utilizes both clinical and research biopsies. A dedicated nurse study coordinator reviews clinical patient list and history on weekly basis and informs the study team of each potential candidate patient. A total of 30 patients having ALK positive lung cancers have been identified and followed up from nearly 500 potentially ALK positive cases between April 2014 to Sept. 2015. Most patients were never smokers but six were former smokers and two were current smokers. Age at diagnoses ranged from 27-78 years (median = 61). Seven patients (23%) were 45 years or younger at the time of diagnosis. A total of 22 patients are currently being treated with crizotinib while eight are on ceritinib or alectinib. For each biopsy, tumor tissues are obtained using a 20 gauge needle, transferred on ice in preserving media and implanted within one hour into 6-8 week old NOD/SCID mice. Many patients have been on treatment for 2-3 years with stable disease so they do not require biopsy. Using a similar strategy, we also obtained biopsies from patients with EGFR gene mutations in their tumors and have progressed while on targeted therapy.

A total of seven cases have been implanted. We will report our experiences in patient selection, clinical follow up, patient consent, PDX development, time from biopsy to tumor establishment, and the results of molecular analyses. Our study enabled us to gain new insights regarding the molecular changes associated with recurring tumors after they have failed targeted therapy as well as clinical experiences on how to utilize state-of-the art approaches and comprehensive genomic information to further improve cancer patient care upon disease progression.

Acknowledgements: This work is supported in part by the Hillsberg Award from the National Foundation for Cancer Research and by the Biomarker Discovery Program at the Mayo Clinic Center for Individualized Medicine.

Citation Format: Jin Jen, Aaron Mansfield, Patrick W. Eiken, Shawn Stoddard, Karlyn Pierson, Xiaonan Hou, Hong Zheng Ren, Julian Molina, Joanne Yi, Ping Yang. Integrated Approaches to Treating Lung Adenocarcinoma Resistant to Targeted Therapy. [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 B34.

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

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

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

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

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