Armored CAR T cells enhance antitumor efficacy and overcome the tumor microenvironment

Scientific Reports - Tập 7 Số 1
Oladapo O. Yeku1, Terence J. Purdon1, Mythili Koneru1, David R. Spriggs1, Renier J. Brentjens1
1Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York 10065, USA

Tóm tắt

AbstractChimeric antigen receptor (CAR) T cell therapy has shown limited efficacy for the management of solid tumor malignancies. In ovarian cancer, this is in part due to an immunosuppressive cytokine and cellular tumor microenvironment which suppresses adoptively transferred T cells. We engineered an armored CAR T cell capable of constitutive secretion of IL-12, and delineate the mechanisms via which these CAR T cells overcome a hostile tumor microenvironment. In this report, we demonstrate enhanced proliferation, decreased apoptosis and increased cytotoxicity in the presence of immunosuppressive ascites. In vivo, we show enhanced expansion and CAR T cell antitumor efficacy, culminating in improvement in survival in a syngeneic model of ovarian peritoneal carcinomatosis. Armored CAR T cells mediated depletion of tumor associated macrophages and resisted endogenous PD-L1-induced inhibition. These findings highlight the role of the inhibitory microenvironment and how CAR T cells can be further engineered to maintain efficacy.

Từ khóa


Tài liệu tham khảo

Davila, M. L. et al. Chimeric antigen receptors for the adoptive T cell therapy of hematologic malignancies. Int. J. Hematol. 99, 361–71 (2014).

Park, J. R. et al. Adoptive transfer of chimeric antigen receptor re-directed cytolytic T lymphocyte clones in patients with neuroblastoma. Mol. Ther. 15, 825–33 (2007).

Feng, K. et al. Chimeric antigen receptor-modified T cells for the immunotherapy of patients with EGFR-expressing advanced relapsed/refractory non-small cell lung cancer. Sci. China. Life Sci. 59, 468–79 (2016).

Beatty, G. L. et al. Mesothelin-specific chimeric antigen receptor mRNA-engineered T cells induce anti-tumor activity in solid malignancies. Cancer Immunol. Res. 2, 112–20 (2014).

Kershaw, M. H. et al. A phase I study on adoptive immunotherapy using gene-modified T cells for ovarian cancer. Clin. Cancer Res. 12, 6106–15 (2006).

Junghans, R. P. et al. Phase I Trial of Anti-PSMA Designer CAR-T Cells in Prostate Cancer: Possible Role for Interacting Interleukin 2-T Cell Pharmacodynamics as a Determinant of Clinical Response. Prostate 76, 1257–70 (2016).

Hwu, P. et al. Lysis of ovarian cancer cells by human lymphocytes redirected with a chimeric gene composed of an antibody variable region and the Fc receptor gamma chain. J. Exp. Med. 178, 361–6 (1993).

Hwu, P. et al. In vivo antitumor activity of T cells redirected with chimeric antibody/T-cell receptor genes. Cancer Res. 55, 3369–73 (1995).

Kim, S., Kim, B. & Song, Y. S. Ascites modulates cancer cell behavior, contributing to tumor heterogeneity in ovarian cancer. Cancer Sci. 107, 1173–8 (2016).

Chen, L. et al. Evaluation of immune inhibitory cytokine profiles in epithelial ovarian carcinoma. J. Obstet. Gynaecol. Res. 35, 212–8 (2009).

Zhao, X., Ye, F., Chen, L., Lu, W. & Xie, X. Human epithelial ovarian carcinoma cell-derived cytokines cooperatively induce activated CD4+ CD25-CD45RA+ naïve T cells to express forkhead box protein 3 and exhibit suppressive ability in vitro. Cancer Sci. 100, 2143–51 (2009).

Li, X. et al. Human ovarian carcinoma cells generate CD4(+)CD25(+) regulatory T cells from peripheral CD4(+)CD25(−) T cells through secreting TGF-beta. Cancer Lett. 253, 144–53 (2007).

Sinha, P., Clements, V. K., Bunt, S. K., Albelda, S. M. & Ostrand-Rosenberg, S. Cross-talk between myeloid-derived suppressor cells and macrophages subverts tumor immunity toward a type 2 response. J. Immunol. 179, 977–83 (2007).

Bak, S. P., Alonso, A., Turk, M. J. & Berwin, B. Murine ovarian cancer vascular leukocytes require arginase-1 activity for T cell suppression. Mol. Immunol. 46, 258–68 (2008).

Quatromoni, J. G. & Eruslanov, E. Tumor-associated macrophages: function, phenotype, and link to prognosis in human lung cancer. Am. J. Transl. Res. 4, 376–89 (2012).

Auer, K. et al. Role of the immune system in the peritoneal tumor spread of high grade serous ovarian cancer. Oncotarget 7, 61336–61354 (2016).

Liu, Y. et al. Regulation of arginase I activity and expression by both PD-1 and CTLA-4 on the myeloid-derived suppressor cells. Cancer Immunol. Immunother. 58, 687–97 (2009).

Liu, Y., Zeng, B., Zhang, Z., Zhang, Y. & Yang, R. B7-H1 on myeloid-derived suppressor cells in immune suppression by a mouse model of ovarian cancer. Clin. Immunol. 129, 471–81 (2008).

Wang, X., Teng, F., Kong, L. & Yu, J. PD-L1 expression in human cancers and its association with clinical outcomes. Onco. Targets. Ther. 9, 5023–39 (2016).

Hamanishi, J. et al. The comprehensive assessment of local immune status of ovarian cancer by the clustering of multiple immune factors. Clin. Immunol. 141, 338–47 (2011).

Sadelain, M., Brentjens, R. & Rivière, I. The basic principles of chimeric antigen receptor design. Cancer Discov. 3, 388–98 (2013).

Zhao, Y. et al. A herceptin-based chimeric antigen receptor with modified signaling domains leads to enhanced survival of transduced T lymphocytes and antitumor activity. J. Immunol. 183, 5563–74 (2009).

Zhong, X.-S., Matsushita, M., Plotkin, J., Riviere, I. & Sadelain, M. Chimeric antigen receptors combining 4-1BB and CD28 signaling domains augment PI3kinase/AKT/Bcl-XL activation and CD8+ T cell-mediated tumor eradication. Mol. Ther. 18, 413–20 (2010).

Wilkie, S. et al. Retargeting of human T cells to tumor-associated MUC1: the evolution of a chimeric antigen receptor. J. Immunol. 180, 4901–9 (2008).

Yeku, O. O. & Brentjens, R. J. Armored CAR T-cells: utilizing cytokines and pro-inflammatory ligands to enhance CAR T-cell anti-tumour efficacy. Biochem. Soc. Trans. 44, 412–8 (2016).

Zhao, J., Zhao, J. & Perlman, S. Differential effects of IL-12 on Tregs and non-Treg T cells: roles of IFN-γ, IL-2 and IL-2R. PLoS One 7, e46241 (2012).

Chmielewski, M., Kopecky, C., Hombach, A. A. & Abken, H. IL-12 Release by Engineered T Cells Expressing Chimeric Antigen Receptors Can Effectively Muster an Antigen-Independent Macrophage Response on Tumor Cells That Have Shut Down Tumor Antigen Expression. Cancer Res. 71, 5697–5706 (2011).

Kerkar, S. P. et al. IL-12 triggers a programmatic change in dysfunctional myeloid-derived cells within mouse tumors. J. Clin. Invest. 121, 4746–57 (2011).

Chinnasamy, D. et al. Local delivery of interleukin-12 using T cells targeting VEGF receptor-2 eradicates multiple vascularized tumors in mice. Clin. Cancer Res. 18, 1672–83 (2012).

Atkins, M. B. et al. Phase I evaluation of intravenous recombinant human interleukin 12 in patients with advanced malignancies. Clin. Cancer Res. 3, 409–17 (1997).

Leonard, J. P. et al. Effects of single-dose interleukin-12 exposure on interleukin-12-associated toxicity and interferon-gamma production. Blood 90, 2541–8 (1997).

Hurteau, J. A., Blessing, J. A., DeCesare, S. L. & Creasman, W. T. Evaluation of recombinant human interleukin-12 in patients with recurrent or refractory ovarian cancer: a gynecologic oncology group study. Gynecol. Oncol. 82, 7–10 (2001).

Lenzi, R. et al. Phase II study of intraperitoneal recombinant interleukin-12 (rhIL-12) in patients with peritoneal carcinomatosis (residual disease < 1 cm) associated with ovarian cancer or primary peritoneal carcinoma. J. Transl. Med. 5, 66 (2007).

Kerkar, S. P. et al. Tumor-specific CD8+ T cells expressing interleukin-12 eradicate established cancers in lymphodepleted hosts. Cancer Res. 70, 6725–34 (2010).

Zhang, L. et al. Tumor-infiltrating lymphocytes genetically engineered with an inducible gene encoding interleukin-12 for the immunotherapy of metastatic melanoma. Clin. Cancer Res. 21, 2278–88 (2015).

Chekmasova, A. A. et al. Successful eradication of established peritoneal ovarian tumors in SCID-Beige mice following adoptive transfer of T cells genetically targeted to the MUC16 antigen. Clin. Cancer Res. 16, 3594–606 (2010).

Koneru, M., Purdon, T. J., Spriggs, D., Koneru, S. & Brentjens, R. J. IL-12 secreting tumor-targeted chimeric antigen receptor T cells eradicate ovarian tumors in vivo. Oncoimmunology 4, e994446 (2015).

Pegram, H. J. et al. Tumor-targeted T cells modified to secrete IL-12 eradicate systemic tumors without need for prior conditioning. Blood 119, 4133–41 (2012).

Lisiero, D. N., Soto, H., Liau, L. M. & Prins, R. M. Enhanced Sensitivity to IL-2 Signaling Regulates the Clinical Responsiveness of IL-12-Primed CD8+ T Cells in a Melanoma Model. doi.org 186, 5068–5077 (2011).

Zhang, X. & Starnbach, M. N. An Excess of the Proinflammatory Cytokines IFN-γ and IL-12 Impairs the Development of the Memory CD8+ T Cell Response to Chlamydia trachomatis. J. Immunol. 195, 1665–75 (2015).

Hodge, D. R., Hurt, E. M. & Farrar, W. L. The role of IL-6 and STAT3 in inflammation and cancer. Eur. J. Cancer 41, 2502–12 (2005).

Duluc, D. et al. Tumor-associated leukemia inhibitory factor and IL-6 skew monocyte differentiation into tumor-associated macrophage-like cells. Blood 110, 4319–30 (2007).

Shanker, A., Singh, S. M. & Sodhi, A. Impairment of T-cell functions with the progressive ascitic growth of a transplantable T-cell lymphoma of spontaneous origin. FEMS Immunol. Med. Microbiol. 27, 247–55 (2000).

Platten, M., von K Doeberitz, N., Oezen, I., Wick, W. & Ochs, K. Cancer Immunotherapy by Targeting IDO1/TDO and Their Downstream Effectors. Front. Immunol. 5, 673 (2014).

Vig, M. et al. Inducible nitric oxide synthase in T cells regulates T cell death and immune memory. J. Clin. Invest. 113, 1734–42 (2004).

Oelkrug, C. & Ramage, J. M. Enhancement of T cell recruitment and infiltration into tumours. Clin. Exp. Immunol. 178, 1–8 (2014).

Colvin, E. K. Tumor-associated macrophages contribute to tumor progression in ovarian cancer. Front. Oncol. 4, 137 (2014).

Watkins, S. K., Egilmez, N. K., Suttles, J. & Stout, R. D. IL-12 rapidly alters the functional profile of tumor-associated and tumor-infiltrating macrophages in vitro and in vivo. J. Immunol. 178, 1357–62 (2007).

Dalton, J. E., Howell, G., Pearson, J., Scott, P. & Carding, S. R. Fas-Fas ligand interactions are essential for the binding to and killing of activated macrophages by gamma delta T cells. J. Immunol. 173, 3660–7 (2004).

Abiko, K. et al. IFN-γ from lymphocytes induces PD-L1 expression and promotes progression of ovarian cancer. Br. J. Cancer 112, 1501–9 (2015).

Abiko, K. et al. PD-L1 on tumor cells is induced in ascites and promotes peritoneal dissemination of ovarian cancer through CTL dysfunction. Clin. Cancer Res. 19, 1363–74 (2013).

Lenzi, R. et al. Phase I study of intraperitoneal recombinant human interleukin 12 in patients with Müllerian carcinoma, gastrointestinal primary malignancies, and mesothelioma. Clin. Cancer Res. 8, 3686–95 (2002).

Grohmann, U. et al. Positive regulatory role of IL-12 in macrophages and modulation by IFN-gamma. J. Immunol. 167, 221–7 (2001).

Doench, J. G. et al. Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Nat. Biotechnol. 34, 184–91 (2016).