Patient-derived melanoma organoid models facilitate the assessment of immunotherapies
Lingling Ou, Shujing Liu, Huaishan Wang, Yeye Guo, Lei Guan, Longbin Shen, Ruhui Luo, David E. Elder, Alexander C. Huang, Giorgos C. Karakousis, John T. Miura, Tara C. Mitchell, Lynn M. Schuchter, Ravi K. Amaravadi, Ahron Flowers, Haiwei Mou, Yi Fan, Wei Guo, Jina Ko, Qing Chen, Bin Tian, Meenhard Herlyn, Xiaowei Xu
Abstract
Background Only a minority of melanoma patients experience durable responses to immunotherapies due to inter- and intra-tumoral heterogeneity in melanoma. As a result, there is a pressing need for suitable preclinical models to investigate resistance mechanisms and enhance treatment efficacy. Methods Here, we report two different methods for generating melanoma patient-derived organoids (MPDOs), one is embedded in collagen gel, and the other is inlaid in Matrigel. MPDOs in Matrigel are used for assessing the therapeutic effects of anti-PD-1 antibodies (αPD-1), autochthonous tumor infiltrating lymphocytes (TILs), and small molecule compounds. MPDOs in collagen gel are used for evaluating the chemotaxis and migratory capacity of TILs. Finding The MPDOs in collagen gel and Matrigel have similar morphology and immune cell composition to their parental melanoma tissues. MPDOs show inter- and intra-tumoral heterogeneity and contain diverse immune cells such as CD4 + , CD8 + T, Treg, CD14 + monocytic, CD15 + , and CD11b + myeloid cells. The tumor microenvironment (TME) in MPDOs is highly immunosuppressive, and the lymphoid and myeloid lineages express similar levels of PD-1, PD-L1, and CTLA-4 as their parental melanoma tissues. Anti-PD-1 antibodies (αPD-1) reinvigorate CD8 + T cells and induce melanoma cell death in the MPDOs. TILs expanded by IL-2 and αPD-1 show significantly lower expression of TIM-3, better migratory capacity and infiltration of autochthonous MPDOs, and more effective killing of melanoma cells than TILs expanded by IL-2 alone or IL-2 with αCD3. A small molecule screen discovers that Navitoclax increases the cytotoxicity of TIL therapy. Interpretation MPDOs may be used to test immune checkpoint inhibitors and cellular and targeted therapies. Funding This work was supported by the NIH grants CA114046, CA261608, CA258113, and the Tara Miller Melanoma Foundation.