Abstract IA04: Overcoming immune resistance with rationally designed combination immunotherapy
Authors: Zappasodi Roberta , Budhu Sadna , Sirard Cynthia , Qi Jingjing , Liu Cailian , Li Yanyun , Senbabaoglu Yasin , Manne Sasikanth , Gasmi Billel , Zhong Hong , Yang Xia , Abu-Akeel Moshen , Schaer David , Huang Alexander , Newman Walter , Wong Philip , Panageas Katherine S. , Postow Michael A. , Koon Henry , Velcheti Vamsidhar , Callahan Margaret K. , Hellmann Matthew D. , Wherry E. John , Merghoub Taha , Wolchok Jedd D. 
Published: June 2022

Abstract
Over the past decade, blockade of the immune checkpoints CTLA-4, PD-1, and PD-L1 has been demonstrated to significantly extend survival of cancer patients across multiple tumor types, including metastatic melanoma, formally proving that immunotherapy is a viable option for the treatment of cancer. These successes have paved the way for the development of additional immune-modulatory antibodies, blocking alternative inhibitory receptors, or engaging costimulatory receptors such as the TNF receptor family member GITR. However, the clinical experience accumulated thus far with checkpoint blockade has clearly shown that only a limited fraction of patients achieve durable clinical benefit with these treatments. This highlights the need to deepen our understanding of the molecular mechanisms underlying response and resistance to immunotherapy and design more personalized and rational combinations based on these therapies. Immune-regulatory mechanisms are one of the major barriers limiting efficacy of immunotherapy. CTLA-4 blockade and GITR costimulation are two immunotherapeutic strategies known to interfere with conventional immunosuppressive regulatory T cells (Tregs). We thus investigated the effects of CTLA-4 blockade and GITR costimulation on suppressive T cells in in vivo mouse melanoma models resistant to these therapies with the aim to clarify the molecular mechanisms underlying refractoriness and provide the rationale to develop more effective therapeutic combinations. To understand the clinical relevance of these findings, we explored the same effects in cancer patients treated with CTLA-4 blockade or GITR costimulation. We found that CTLA-4 blockade, while counteracting conventional Tregs, promotes the expansion of a subset of CD4+Foxp3-T cells expressing high levels of PD-1 (4PD1hi), which constitute a new immunosuppressive T-cell population with T-follicular-helper-like features. Importantly, we observed that anti-CTLA-4 increases the frequency of circulating 4PD1hi in a dose-dependent manner. In contrast, PD-1 blockade decreases 4PD1hi in function of its clinical activity, underscoring the relevance of this cell subset as a pharmacodynamic and prognostic biomarker of checkpoint blockade. These findings indicate that optimizing checkpoint blockade doses and combination regimens so as to keep 4PD1hi in the right balance may favor a positive outcome. In mouse melanoma models of response (early tumors) and refractoriness (advanced tumors) to GITR agonism, we found that anti-GITR efficiently reduces Tregs and increases effector:Treg ratios in both curative and refractory treatment conditions. However, T-cell activation and cytotoxic functions are favored selectively in the presence of low tumor burden. Counteracting exhaustion with PD-1 blockade in combination with GITR agonism restored responsiveness of advanced tumors and CD8+ T-cell functionality. Aligned with the effects of anti-GITR in mice, we found that the agonist anti-human GITR antibody TRX518 decreases Tregs in peripheral blood and tumor to similar extents in patients treated in the first-in-human single-dose monotherapy trial (NCT01239134). However, coincident downregulation of Tregs in the peripheral blood and at the tumor site upon TRX518 was not sufficient to achieve substantial clinical responses in this population of advanced solid cancer patients. This suggests that Treg elimination from advanced tumors may not be sufficient to activate cytotoxic T-cell responses unless the T-cell exhaustion process is concurrently blocked. Based on these preclinical and clinical observations, we have started to explore anti-GITR in combination with PD-1 pathway blockade in patients with advanced solid tumors (NCT02628574). Taken as a whole, these findings illustrate the value of conventional and nonconventional immune-suppressive T cells as biomarkers of biologic and therapeutic activity of immunotherapy in melanoma and other tumor types. In addition, these results indicate that inhibition of immune-regulatory mechanisms, such as immunosuppressive T cells, may need to be associated with strategies able to positively activate T-cell responses to achieve significant clinical benefit.
Citation Format: Roberta Zappasodi, Sadna Budhu, Cynthia Sirard, Jingjing Qi, Cailian Liu, Yanyun Li, Yasin Senbabaoglu, Sasikanth Manne, Billel Gasmi, Hong Zhong, Xia Yang, Moshen Abu-Akeel, David Schaer, Alexander Huang, Walter Newman, Philip Wong, Katherine S. Panageas, Michael A. Postow, Henry Koon, Vamsidhar Velcheti, Margaret K. Callahan, Matthew D. Hellmann, E. John Wherry, Taha Merghoub, Jedd D. Wolchok. Overcoming immune resistance with rationally designed combination immunotherapy [abstract]. In: Proceedings of the AACR Special Conference on Melanoma: From Biology to Target; 2019 Jan 15-18; Houston, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(19 Suppl):Abstract nr IA04.