Cancer Therapy : Preclinical Immunotherapy of Metastatic Melanoma Using Genetically Engineered GD 2-Specific T cells

Purpose: Genetic engineering of human T lymphocytes to express tumor-directed chimeric antigen receptors (CAR) can produce antitumor effector cells that bypass tumor immune escape mechanisms that are due to abnormalities in protein-antigen processing and presentation. Moreover, these transgenic receptors can be directed to tumor-associated antigens that are not protein-derived, such as the ganglioside GD2, which is expressed in a high proportion of melanoma cells. Experimental Design: We generated chimeric T cells specific for the ganglioside GD2 by joining an extracellular antigen-binding domain derived from the GD2-specific antibody sc14.G2a to cytoplasmic signaling domains derived from the T-cell receptor ζ-chain, with the endodomains of the costimulatory molecules CD28 and OX40. We expressed this CAR in human T cells and assessed the targeting of GD2-positive melanoma tumors in vitro and in a murine xenograft. Results: Upon coincubation with GD2-expressing melanoma cells, CAR-GD2 T lymphocytes incorporating the CD28 and OX40 endodomains secreted significant levels of cytokines in a pattern comparable with the cytokine response obtained by engagement of the native CD3 receptor. These CAR-T cells had antimelanoma activity in vitro and in our xenograft model, increasing the survival of tumor-bearing animals. Conclusion: Redirecting human T lymphocytes to the tumor-associated ganglioside GD2 generates effector cells with antimelanoma activity that should be testable in subjects with disease. (Clin Cancer Res 2009;15(18):5852–60) The rising incidence of cutaneous melanoma and the failure to significantly improve outcomes in metastatic disease have led to increasing interest in immunotherapeutic approaches, because these can be remarkably effective (1–3). Several investigators have focused on targeting tumor-associated antigens that fall into the cancer testis antigen group, including MAGE, BAGE, GAGE, and NY-ESO-1, or the melanocyte differentiation protein group, including gp100, Melan-A/MART-1, and tyrosinase, which are widely present on melanoma cells. These studies have used cytotoxic T cell lines (4, 5), clones with native (6) or transgenic αβ T cell receptors (7) specific for cancer testis antigen–derived peptides that are recognized in association with human leukocyte antigen (HLA) class I antigens on the tumor cell surface. It is clear, however, that the heterogeneity of protein antigen expression and presentation in melanoma is a characteristic that helps limit the proportion of patients who are able to respond to such targeted strategies (8). One means of increasing the effectiveness of targeted T cell therapy of melanoma, therefore, may be to use artificial chimeric receptors derived, for example, from the antigen binding domain of a monoclonal antibody (9). When coupled to appropriate intracellular signaling domains, T cells expressing these chimeric antigen receptors (CAR) can kill tumor cell targets (10). They have the advantage of acting in a MHC unrestricted manner, Authors' Affiliations: Center for Cell and Gene Therapy, Baylor College of Medicine, Texas Children's Hospital, The Methodist Hospital, and Departments of Medicine and Immunology, Baylor College of Medicine, Houston, Texas; Medical Oncology Unit, Fondazione IRCCS Istituto Nazionale per lo Studio e la Cura dei Tumori, and Tumor Immunobiology Unit, Istituto Nazionale Tumori, Milan, Italy; and INSERM-U590, Centre Léon-Bérard, Lyon, France Received 12/5/08; revised 5/5/09; accepted 5/19/09; published OnlineFirst 9/8/09. Grant support: NIH grant PO1 CA94237. B. Savoldo is supported by NIH grant 1RO1CA131027 and by a Leukemia and Lymphoma Society Translational Research grant. G. Dotti is supported by the Doris Duke Charitable Foundation/Clinical Scientist development award and by a Leukemia and Lymphoma Society Translational Research grant. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Note: Supplementary data for this article are available at Clinical Cancer Research Online (http://clincancerres.aacrjournals.org/). Authorship: E. Yvon, M. Del Vecchio, B. Savoldo, V.Hoyos, and A. Dutour designed and did the experiments. E. Yvon, B. Savoldo, G. Dotti, and M.K. Brenner designed the research and analyzed the data. E. Yvon, G. Dotti, andM.K. Brennerwrote themanuscript. A. Anichini provided critical revision of the manuscript. All authors approved the final version of the manuscript. Requests for reprints: Eric Yvon, Center for Cell and Gene Therapy, Baylor College of Medicine, 6621 Fannin Street MC 3-3320, Houston, TX 77030. Phone: 832-824-4549; Fax: 832-825-4732; E-mail: exyvon@txccc.org. F 2009 American Association for Cancer Research. doi:10.1158/1078-0432.CCR-08-3163 5852 Clin Cancer Res 2009;15(18) September 15, 2009 www.aacrjournals. org Research. on June 29, 2017. © 2009 American Association for Cancer clincancerres.aacrjournals. org Downloaded from Published OnlineFirst September 8, 2009; DOI: 10.1158/1078-0432.CCR-08-3163