Single-agent immunotherapy has achieved limited clinical benefit to date in patients with pancreatic ductal adenocarcinoma (PDAC). This may be a result of the presence of a uniquely immunosuppressive tumor microenvironment (TME). Critical obstacles to immunotherapy in PDAC tumors include a high number of tumor-associated immunosuppressive cells and a uniquely desmoplastic stroma that functions as a barrier to T cell infiltration. We identified hyperactivated focal adhesion kinase (FAK) activity in neoplastic PDAC cells as an important regulator of the fibrotic and immunosuppressive TME. We found that FAK activity was elevated in human PDAC tissues and correlated with high levels of fibrosis and poor CD8+ cytotoxic T cell infiltration. Single-agent FAK inhibition using the selective FAK inhibitor VS-4718 substantially limited tumor progression, resulting in a doubling of survival in the p48-Cre;LSL-KrasG12D;Trp53flox/+ (KPC) mouse model of human PDAC. This delay in tumor progression was associated with markedly reduced tumor fibrosis and decreased numbers of tumor-infiltrating immunosuppressive cells. We also found that FAK inhibition rendered the previously unresponsive KPC mouse model responsive to T cell immunotherapy and PD-1 antagonists. These data suggest that FAK inhibition increases immune surveillance by overcoming the fibrotic and immunosuppressive PDAC TME and renders tumors responsive to immunotherapy.
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Royal, R.E. et al. Phase 2 trial of single agent Ipilimumab (anti-CTLA-4) for locally advanced or metastatic pancreatic adenocarcinoma. J. Immunother. 33, 828–833 (2010).
Zhu, Y. et al. CSF1/CSF1R blockade reprograms tumor-infiltrating macrophages and improves response to T cell checkpoint immunotherapy in pancreatic cancer models. Cancer Res. 74, 5057–5069 (2014).
Panni, R.Z., Linehan, D.C. & DeNardo, D.G. Targeting tumor-infiltrating macrophages to combat cancer. Immunotherapy 5, 1075–1087 (2013).
Mitchem, J.B. et al. Targeting tumor-infiltrating macrophages decreases tumor-initiating cells, relieves immunosuppression and improves chemotherapeutic responses. Cancer Res. 73, 1128–1141 (2013).
Goedegebuure, P. et al. Myeloid-derived suppressor cells: general characteristics and relevance to clinical management of pancreatic cancer. Curr. Cancer Drug Targets 11, 734–751 (2011).
Bayne, L.J. et al. Tumor-derived granulocyte–macrophage colony-stimulating factor regulates myeloid inflammation and T cell immunity in pancreatic cancer. Cancer Cell 21, 822–835 (2012).
Laklai, H. et al. Genotype tunes pancreatic ductal adenocarcinoma tissue tension to induce matricellular fibrosis and tumor progression. Nat. Med. 22, 497–505 (2016).
Feig, C. et al. Targeting CXCL12 from FAP-expressing carcinoma-associated fibroblasts synergizes with anti-PD-L1 immunotherapy in pancreatic cancer. Proc. Natl. Acad. Sci. USA 110, 20212–20217 (2013).
Özdemir, B.C. et al. Depletion of carcinoma-associated fibroblasts and fibrosis induces immunosuppression and accelerates pancreas cancer with reduced survival. Cancer Cell 25, 719–734 (2014).
Olive, K.P. et al. Inhibition of Hedgehog signaling enhances delivery of chemotherapy in a mouse model of pancreatic cancer. Science 324, 1457–1461 (2009).
Provenzano, P.P. et al. Enzymatic targeting of the stroma ablates physical barriers to treatment of pancreatic ductal adenocarcinoma. Cancer Cell 21, 418–429 (2012).
Beatty, G.L. et al. Exclusion of T cells from pancreatic carcinomas in mice is regulated by Ly6ClowF4/80+ extratumoral macrophages. Gastroenterology 149, 201–210 (2015).
Sanford, D.E. et al. Inflammatory monocyte mobilization decreases patient survival in pancreatic cancer: a role for targeting the CCL2–CCR2 axis. Clin. Cancer Res. 19, 3404–3415 (2013).
Sulzmaier, F.J., Jean, C. & Schlaepfer, D.D. FAK in cancer: mechanistic findings and clinical applications. Nat. Rev. Cancer 14, 598–610 (2014).
Serrels, A. et al. Nuclear FAK controls chemokine transcription, Tregs and evasion of antitumor immunity. Cell 163, 160–173 (2015).
Tavora, B. et al. Endothelial cell FAK targeting sensitizes tumors to DNA-damaging therapy. Nature 514, 112–116 (2014).
Zhao, X.K. et al. Focal adhesion kinase regulates fibroblast migration via integrin β-1 and plays a central role in fibrosis. Sci. Rep. 6, 19276 (2016).
Balasubramanian, S. et al. Dasatinib attenuates pressure overload induced cardiac fibrosis in a murine transverse aortic constriction model. PLoS One 10, e0140273 (2015).
Rustad, K.C., Wong, V.W. & Gurtner, G.C. The role of focal adhesion complexes in fibroblast mechanotransduction during scar formation. Differentiation 86, 87–91 (2013).
Sonomura, K. et al. The kinase Pyk2 is involved in renal fibrosis by means of mechanical stretch-induced growth factor expression in renal tubules. Kidney Int. 81, 449–457 (2012).
You, K., Huang, Y., Zhang, M.C. & Hao, J. Control and prevention of myocardial fibrosis using Pyk2-related non-kinase. Int. J. Clin. Exp. Med. 8, 18284–18292 (2015).
Koppel, A.C. et al. Delayed skin wound repair in proline-rich protein tyrosine kinase 2–knockout mice. Am. J. Physiol. Cell Physiol. 306, C899–C909 (2014).
Graves, D.T., Wu, Y. & Badadani, M. Pyk2 contributes to re-epithelialization by promoting MMP expression. Focus on 'Delayed skin wound repair in proline-rich protein tyrosine kinase 2–knockout mice'. Am. J. Physiol. Cell Physiol. 306, C887–C888 (2014).
Okigaki, M. et al. Pyk2 regulates multiple signaling events crucial for macrophage morphology and migration. Proc. Natl. Acad. Sci. USA 100, 10740–10745 (2003).
Stokes, J.B. et al. Inhibition of focal adhesion kinase by PF-562,271 inhibits the growth and metastasis of pancreatic cancer concomitant with altering the tumor microenvironment. Mol. Cancer Ther. 10, 2135–2145 (2011).
Konstantinidou, G. et al. RHOA–FAK is a required signaling axis for the maintenance of KRAS-driven lung adenocarcinomas. Cancer Discov. 3, 444–457 (2013).
Bae, Y.H. et al. A FAK–Cas–Rac–lamellipodin signaling module transduces extracellular matrix stiffness into mechanosensitive cell cycling. Sci. Signal. 7, ra57 (2014).
Kümper, S. & Marshall, C.J. ROCK-driven actomyosin contractility induces tissue stiffness and tumor growth. Cancer Cell 19, 695–697 (2011).
Samuel, M.S. et al. Actomyosin-mediated cellular tension drives increased tissue stiffness and β-catenin activation to induce epidermal hyperplasia and tumor growth. Cancer Cell 19, 776–791 (2011).
Wozniak, M.A., Desai, R., Solski, P.A., Der, C.J. & Keely, P.J. ROCK-generated contractility regulates breast epithelial cell differentiation in response to the physical properties of a three-dimensional collagen matrix. J. Cell Biol. 163, 583–595 (2003).
Pylayeva, Y. et al. Ras- and PI3K-dependent breast tumorigenesis in mice and humans requires focal adhesion kinase signaling. J. Clin. Invest. 119, 252–266 (2009).
Rhim, A.D. et al. Stromal elements act to restrain, rather than support, pancreatic ductal adenocarcinoma. Cancer Cell 25, 735–747 (2014).
Rhim, A.D. et al. EMT and dissemination precede pancreatic tumor formation. Cell 148, 349–361 (2012).
Kim, M.P. et al. ALDH activity selectively defines an enhanced tumor-initiating cell population relative to CD133 expression in human pancreatic adenocarcinoma. PLoS One 6, e20636 (2011).
Hermann, P.C. et al. Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 1, 313–323 (2007).
Crompton, B.D. et al. High-throughput tyrosine kinase activity profiling identifies FAK as a candidate therapeutic target in Ewing sarcoma. Cancer Res. 73, 2873–2883 (2013).
DeNardo, D.G. et al. Leukocyte complexity predicts breast cancer survival and functionally regulates response to chemotherapy. Cancer Discov. 1, 54–67 (2011).
Ruffell, B. et al. Macrophage IL-10 blocks CD8+ T cell–dependent responses to chemotherapy by suppressing IL-12 expression in intratumoral dendritic cells. Cancer Cell 26, 623–637 (2014).
Strachan, D.C. et al. CSF1R inhibition delays cervical and mammary tumor growth in murine models by attenuating the turnover of tumor-associated macrophages and enhancing infiltration by CD8+ T cells. OncoImmunology 2, e26968 (2013).
Shree, T. et al. Macrophages and cathepsin proteases blunt chemotherapeutic response in breast cancer. Genes Dev. 25, 2465–2479 (2011).
Beatty, G.L. et al. Mesothelin-specific chimeric antigen receptor mRNA–engineered T cells induce antitumor activity in solid malignancies. Cancer Immunol. Res. 2, 112–120 (2014).
Highfill, S.L. et al. Disruption of CXCR2-mediated MDSC tumor trafficking enhances anti-PD1 efficacy. Sci. Transl. Med. 6, 237ra67 (2014).
Mok, S. et al. Inhibition of CSF-1 receptor improves the antitumor efficacy of adoptive cell transfer immunotherapy. Cancer Res. 74, 153–161 (2014).
Beatty, G.L. et al. CD40 agonists alter tumor stroma and show efficacy against pancreatic carcinoma in mice and humans. Science 331, 1612–1616 (2011).
Chapman, N.M. & Houtman, J.C. Functions of the FAK family kinases in T cells: beyond actin cytoskeletal rearrangement. Immunol. Res. 59, 23–34 (2014).
Chapman, N.M., Connolly, S.F., Reinl, E.L. & Houtman, J.C. Focal adhesion kinase negatively regulates Lck function downstream of the T cell antigen receptor. J. Immunol. 191, 6208–6221 (2013).
Chapman, N.M., Yoder, A.N. & Houtman, J.C. Noncatalytic functions of Pyk2 and Fyn regulate late stage adhesion in human T cells. PLoS One 7, e53011 (2012).
Collins, M., Bartelt, R.R. & Houtman, J.C. T cell receptor activation leads to two distinct phases of Pyk2 activation and actin cytoskeletal rearrangement in human T cells. Mol. Immunol. 47, 1665–1674 (2010).
Stewart, J.E. et al. Inhibition of FAK and VEGFR-3 binding decreases tumorigenicity in neuroblastoma. Mol. Carcinog. 54, 9–23 (2015).
Golubovskaya, V. et al. Downregulation of ALDH1A3, CD44 or MDR1 sensitizes resistant cancer cells to FAK autophosphorylation inhibitor Y15. J. Cancer Res. Clin. Oncol. 141, 1613–1631 (2015).
Zheng, D. et al. A novel strategy to inhibit FAK and IGF-1R decreases growth of pancreatic cancer xenografts. Mol. Carcinog. 49, 200–209 (2010).
Hochwald, S.N. et al. A novel small-molecule inhibitor of FAK decreases growth of human pancreatic cancer. Cell Cycle 8, 2435–2443 (2009).
François, R.A. et al. Targeting focal adhesion kinase and resistance to mTOR inhibition in pancreatic neuroendocrine tumors. J. Natl. Cancer Inst. 107, djv123 (2015).
Kim, M.P. et al. Generation of orthotopic and heterotopic human pancreatic cancer xenografts in immunodeficient mice. Nat. Protoc. 4, 1670–1680 (2009).
Weischenfeldt, J. & Porse, B. Bone marrow–derived macrophages (bmm): isolation and applications. Cold Spring Harb. Protoc. 12, pdb-prot5080 (2008).
Pelham, R.J. Jr. & Wang, Yl. Cell locomotion and focal adhesions are regulated by substrate flexibility. Proc. Natl. Acad. Sci. USA 94, 13661–13665 (1997).
KC (KrasG12D) cells were obtained from P. Mukherjee (University of North Carolina). HPNE, HPAC, Capan-1, Capan-2, Hs766T, MIA PaCa-2 and SW1990 cells were obtained from K. Lim (Washington University). pBABEpuro K-Ras G12V used to express KrasG12V was obtained from J. Weber (Washington University). This work was supported by funding awarded to D.G.D. by Lustgarten Foundation, an AACR/PANCAN Award, NCI awards R01-CA177670, R01-CA203890 and R21-CA182701, the BJCIH/Siteman Cancer Center Cancer Frontier Fund, and Washington University Clinical and Translational Grant KL2TR000450 awarded to A.W.G.
I.M.S., D.T.W. and J.A.P. are employees of Verastem, Inc.
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Jiang, H., Hegde, S., Knolhoff, B. et al. Targeting focal adhesion kinase renders pancreatic cancers responsive to checkpoint immunotherapy. Nat Med 22, 851–860 (2016). https://doi.org/10.1038/nm.4123
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