Abstract 924: FAK inhibition preferentially attenuates growth of Merlin-negative malignant mesotheliomas: role of cancer stem cells.
Authors: Shapiro Irina M. , Kolev Vihren N. , Vidal Christian M. , Ring Jennifer E. , Keegan Mitchell , Xu Qunli , Menges Craig W. , Testa Joseph R. , Pachter Jonathan A. 
Published: August 2014

Abstract
Malignant pleural mesothelioma (MPM) is an aggressive tumor in the pleural lining of the lung often caused by asbestos exposure. MPM patients are usually diagnosed at an advanced stage of the disease and the prognosis is poor. Median survival after diagnosis is 9 to 12 months and standard-of-care agents such as pemetrexed are relatively ineffective in increasing median survival time for MPM patients. New therapeutic modalities are urgently needed for MPM patients.
Neurofibromatosis 2 (NF2) is a tumor suppressor gene that encodes the protein Merlin. Biallelic inactivation of NF2 by mutation and/or deletion occurs in ∼40% of MPMs leading to inactive or absent Merlin. Merlin has been demonstrated to play roles in cell adhesion, invasion and cell motility in tumor cell lines partially through regulation of focal adhesion kinase (FAK) which in turn mediates signal transduction by integrins and growth factor receptors. Increased activation of FAK has been demonstrated in NF2-mutated mesothelioma cells, indicating that FAK may represent an important therapeutic target for MPM.
A potent and selective FAK inhibitor VS-4718 was evaluated in a panel of MPM cell lines with wild-type or mutated NF2. Mutant NF2 MPM cells were found to be especially sensitive to the FAK inhibitor VS-4718 in a 3D Matrigel assay with EC50 values below 100 nM, in contrast to wild type NF2 MPM cell lines which were less sensitive with EC50 values above 1 μM. Decrease in cell growth in response to FAK inhibitor treatment was found to result from reduction of cell proliferation and induction of apoptosis. Ectopic expression of a non-phosphorylatable mutant form of NF2 (NF2-S518A) in NF2 mutant MPM cells abolished the enhanced sensitivity to VS-4718 in both proliferation and apoptosis assays, confirming the hypothesis that Merlin loss confers hypersensitivity to FAK inhibition. In addition, VS-4718 inhibited tumor cell growth in an orthotopic Merlin-negative mesothelioma xenograft model in a dose-dependent manner with corresponding inhibition of tumor FAK autophosphorylation. It has been reported that cancer stem cells (CSCs) are enriched in pemetrexed-resistant mesothelioma. Using Aldefluor activity as a measure of CSCs, we have found that the FAK inhibitor VS-4718 preferentially reduced the percentage of CSCs in Merlin-negative MPM in contrast to the standard-of-care agent pemetrexed which increased the percentage of CSCs. In summary, our results indicate that the FAK inhibitor VS-4718 is especially potent in Merlin-negative MPM tumor cells, and that NF2 status may be a valuable stratification marker for response to FAK inhibition. Furthermore, cancer stem cells in Merlin-negative mesotheliomas appear to be particularly resistant to pemetrexed, but sensitive to VS-4718. These data support the clinical development of a selective FAK inhibitor for treatment of Merlin-negative malignant mesothelioma.
Citation Format: Irina M. Shapiro, Vihren N. Kolev, Christian M. Vidal, Jennifer E. Ring, Mitchell Keegan, Qunli Xu, Craig W. Menges, Joseph R. Testa, Jonathan A. Pachter. FAK inhibition preferentially attenuates growth of Merlin-negative malignant mesotheliomas: role of cancer stem cells. [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 924. doi:10.1158/1538-7445.AM2013-924