Tumor cells can grow in an anchorage-independent manner. This is mediated in part through survival signals that bypass normal growth restraints controlled by integrin cell surface receptors. Focal adhesion kinase (FAK) is a cytoplasmic protein-tyrosine kinase that associates with integrins and modulates various cellular processes including growth, survival, and migration. As increased FAK expression and tyrosine phosphorylation are associated with tumor progression, inhibitors of FAK are being tested for anti-tumor effects. Here, we analyze PND-1186, a substituted pyridine reversible inhibitor of FAK activity with a 50% inhibitory concentration (IC50) of 1.5 nM in vitro. PND-1186 has an IC50 of ~100 nM in breast carcinoma cells as determined by anti-phospho-specific immunoblotting to FAK Tyr-397. PND-1186 did not alter c-Src or p130Cas tyrosine phosphorylation in adherent cells, yet functioned to restrain cell movement. Whereas 1.0 µM PND-1186 (>5-fold above IC50) had limited effects on cell proliferation, under non-adherent conditions or when grown as spheroids or colonies in soft agar, 0.1 µM PND-1186 blocked FAK and p130Cas tyrosine phosphorylation, promoted caspase-3 activation, and triggered cell apoptosis. PND-1186 inhibited 4T1 breast carcinoma subcutaneous tumor growth correlated with elevated tumor cell apoptosis and caspase 3 activation. Addition of PND-1186 to the drinking water of mice was well tolerated and inhibited ascites-associated ovarian carcinoma tumor growth associated with the inhibition of FAK tyrosine phosphorylation. Our results with low-level PND-1186 treatment support the conclusion that FAK activity selectively promotes tumor cell survival in three-dimensional environments.

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Background: While the human tumor xenograft models established by inoculation of human cancer cell lines into immunodeficient mice have been widely used for test of novel cytotoxic anticancer agents, new drug development has moved from general cytotoxic agents to molecular target-directed therapeutics. Consequently, there is a need to identify tumor types and individual patient tumors that express the target and could benefit from more selective therapies in clinical trials. Therefore, the in vivo models used in preclinical development should be “disease-oriented” and target-directed. Recently, we developed xenograft tumor models by transplanting human fresh tumor fragments into nude mice, which have been used for test of clinically used and novel anticancer drugs. Methods: The fresh tumor samples were collected from local hospitals. The tumor fragments of 1–2 mm were subcutaneously implanted in the flanks of the Balb/c nude mice. In the first passage, tumors derived from male patients were implanted into male mice, and tumors from women were inoculated into female mice. The histology and genomic mutation status were compared between original patients' tumors and the genografts. All therapeutic efficacy experiments, with the exception of prostate cancer, used female mice. The positive control drugs tested included cisplatin, paclitaxel, docetaxel, irinotecan, doxorubicin, 5-FU, gemcitabine, and erlotinib. Results: A total of 537 human tumor samples have been implanted into nude mice, 221 patient tumor-derived models have been established. The tumor taking rates of the first passage were colorectal (69%), ovarian (64%), esophagel (63%), small cell lung cancer (60%), non-small cell lung cancer (54%), gastric (25%), kidney (17%), glioblastoma (16%), breast (12%), liver (12%), and acute lymphocytic leukemia (25%). The tumor taking rates were higher in the later passages for the various tumor types, ranged from approximately 50–100%. The test anticancer drugs produced tumor inhibition rates ranged from 20–90%, which were consistent with their clinical findings. The patient-tumor xenografts from all five generations presented the same histopathological morphology and genomic mutation status to their counterparts of the human primary tumors. Conclusions: These results suggest that patient-tumor derived xenograft tumor models provide a unique renewable source of tumor material for test of novel anticancer agents and may give a better predictive value than the traditional human tumor xenograft models established by inoculation of cancer cell lines. Especially, they have advantages for test of target-oriented therapeutics in new drugs development programs. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr A19.

BACKGROUND Human tumor xenograft models established by transplantation of human tumor cell lines into immunodifficient mice have been routinely used for preclinical test of anticancer agents. But tumor cell lines have a relatively low transplantability and resulted in a limited number of tumor models available for selection of right models for testing novel agents based on their antitumor mechanism. Recently, we have developed a large number of patient primary non-small cell lung cancer (NSCLC) xenograft models by transplanting patients’ fresh tumor tissues into nude mice, which have been employed for preclinical test of anticancer agents. METHODS The fresh NSCLC tissues were collected from local hospitals. The tumor fragments of 1-2 mm were subcutaneously implanted in the flanks of the nude mice by trocar needle. Sixteen tumor fragments were grafted into four mice from one patient tumor tissue (passage 0). All therapeutic efficacy experiments used female mice bearing passage 5 xenografts. The test drugs included cisplatin, carboplatin, paclitaxel, docetaxel, gemcitabine, erlotinib, gefitinib, and pemetrexed. RESULTS A total of 213 patient primary NSCLC samples were implanted into nude mice, and 107 primary tumor models have been established with a tumor taking rate of 50% for the first passage. The tumor taking rates were higher (80-100) in the later passages. The therapeutic efficacy of the test drugs in these models is consistent with their clinical findings. The histopathology and gene sequence of the established primary tumor xenografts were analyzed; their architecture, histopathological morphology, and genomic mutation status from five generations of xenografts retained the patients’ original tumor characteristics. CONCLUSIONS The patient primary NSCLC models can be established in a large number for right models selection in preclinical setting, and have been employed for test of standard of care drugs and novel anticancer agents. The primary tumor models retain a similarity in histology and genomic mutation status to their patients’ original tumors. They may predict more relevant clinical response rate and higher correlation with clinical findings than use of traditional xenograft models established from long-term cultured cancer cell lines. Especially, they have advantages for test of target-oriented therapeutics in new drugs discovery and development programs. Citation Format: Changnian Liu, Connie Sun, Wenwei Li, Wen Zhou, Yong Liu, Rui Zhou, Fang He, Chang Bai. Establishment of patient non-small cell lung cancer derived models for test of anticancer drugs. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2776. doi:10.1158/1538-7445.AM2013-2776