Cancer immunotherapy has transformed the treatment landscape for a number of cancer patients, with some achieving durable and long lasting clinical benefit. Cancer immunotherapy engages and intensifies the host immune response to attack and kill tumor cells. However, as evidenced by the heterogeneous response to immunotherapy, tumor cells have evolved a host of known and unknown mechanisms to evade, inhibit or supersede the immune response. Consequently, scientists and clinicians are unable to accurately predict which patients will respond, or how well they will respond to cancer immunotherapy.To address this shortfall, we have asked the question of how we can modulate tumor cells in order to make them more amenable to immunotherapy, thereby increasing its efficacy. We approached this question by conducting a high throughput drug screen of 850 compounds, to identify bioactive drugs that can increase T cell mediated killing of tumor cells. The goal here is to develop rational combination treatment strategies involving T cell based cancer immunotherapy that will increase the breadth and depth of the clinical response to cancer immunotherapy. One of three top hits from the screen was Topoisomerase I (Top1) inhibitors including irinotecan, topotecan, and camptothecin. We then utilized multiple patient-derived cell lines in an in vitro cytotoxicity assay to validate that treatment of melanoma tumor cells with a Top1 inhibitor, before incubation with their autologous tumor infiltrating lymphocytes (TILs) results in a synergistic increase in T cell mediated killing of tumor cells.These findings were further corroborated in a pre-clinical mouse model, where we found that tumor-bearing mice treated with a combination of a clinically relevant Top1 inhibitor nal-IRI (nano-liposomal irinotecan) and an anti-PD-L1 antibody, showed enhanced tumor regression compared to mice treated with either single agent (mean tumor volume: combo vs nal-IRI vs α-PDL1 = 40.04 ± 5.66 vs 136.30 ± 28.96 vs 373.04 ± 23.96 mm3 respectively, on day 21 after tumor inoculation, p < 0.0001). Significantly longer survival was also achieved in tumor-bearing mice treated with the combination in comparison to cohorts treated with either single agent. To investigate the molecular changes being mediated by Top1 inhibitors in the tumor cells, we conducted gene expression analysis on Top1 inhibitor-treated tumor cells. One striking gene expression change in Top1 inhibitor-treated tumor cells was an upregulation of a number of genes known to be functionally important for p53 signaling including TP53INP1 (Teap). We then focused on the functional relevance of Teap to the increased T cell mediated killing of Top1 inhibitor-treated melanoma cells. Overexpression of Teap in melanoma cells resulted in increased T cell mediated killing, recapitulating the phenotype observed in Top1 inhibitor-treated melanoma cells. Complementary to this, silencing of Teap via shRNA in melanoma cells, inhibited T cell mediated killing of Top1 inhibitor-treated cells, indicating that the enhancement of T cell mediated killing observed in Top1 inhibitor-treated cells is dependent on the p53 regulatory gene Teap. These results support our goal of developing combinations involving T cell based cancer immunotherapy to improve therapeutic efficacy in cancer patients. We have demonstrated that Top1 inhibitors can be effectively combined with T cell based cancer immunotherapy. The results are also indicative of a role for p53 signaling in regulating response to T cell based immunotherapy. By understanding the molecular mechanisms in the tumor that can dictate response or resistance to immunotherapy, we can develop a more comprehensive picture of the cancer immunity response cycle and develop more effective strategies to combat not only melanoma, but also other tumor types where immunotherapy is not yet applicable.
Citation Format: Jodi A. McKenzie, Rina M. Mbofung, Shruti Malu, Rodabe N. Amaria, Emily L. Ashkin, Seram N. Devi, Weiyi Peng, Leila J. Williams, Richard E. Davis, Jason Roszik, Trang N. Tieu, Timothy Heffernan, Patrick Hwu. Topoisomerase I inhibitors enhance efficacy of immunotherapy through a p53 regulatory pathway [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B110.
- ©2016 American Association for Cancer Research.