Targeting autophagy to enhance immune checkpoint inhibition

项目摘要-项目2 一种可以克服对免疫检查点抑制（ICI）的抗性的新方法是一个主要的未满足的需求， 四期黑色素瘤患者。本项目的总体目标是确定ICI与自噬的结合是否 抑制可以解决这种未满足的需求。自噬是化疗的一种关键耐药机制， 疗法最近，我们的工作和其他人的工作表明自噬是一种抵抗机制， 免疫疗法这提出了一些问题，即哪种方法是靶向自噬的最佳方法， 在肿瘤微环境中，哪些细胞类型对靶向自噬最关键。该项目将 利用与新兴生物技术公司的深入合作，这些公司已经开发出下一代 化学的溶酶体和非溶酶体的自噬抑制剂，正走向临床。以下目标将 检验我们的总体假设，即溶酶体自噬抑制导致集中的细胞通路扰动 在癌细胞和免疫细胞中增强ICI的功效：特异性目标1将决定其机制 通过新的临床级自噬抑制剂调节ICI期间的肿瘤-免疫相互作用，重点关注 对骨髓、肿瘤、T细胞和其他免疫细胞表型的影响。我们将比较每一个的能力， 在临床相关的小鼠模型中，抗PD-1和抗CTLA-4抗体的联合使用可增强抗PD-1和抗CTLA-4抗体的效果。我们将重点 PPT 1是一种调节自噬的溶酶体硫酯酶，也是氯喹的主要分子靶点 衍生物，以及癌症药物开发的令人兴奋的新靶点。我们将利用我们的新条件Ppt 1 KO小鼠模型，以比较ICI与Ppt 1 KO组合在肿瘤细胞、树突状细胞和骨髓中的作用。 细胞，并比较遗传抑制与化学Ppt 1抑制对黑色素瘤肿瘤生长的影响。具体目标2 将确定TME的免疫特征在患者和临床前模型中的变化， ICI和自噬抑制。我们将进行LIMIT黑色素瘤试验，这是纳武单抗的适应性I/II期试验， + HCQ和纳武单抗+伊匹单抗+ HCQ在IV期黑素瘤患者中的应用。新型PET成像技术 将用于跟踪肿瘤中的CD 8 + T细胞，并将CD 8+信号与组织学CD 8+分析相关联， 临床反应。在相关的临床前小鼠研究中，我们将使用无偏倚的方法来实现 在早期使用ICI和HCQ或ICI和DC 661治疗的黑色素瘤肿瘤中的变化的综合观点 反应和抵抗。影响：我们的研究将确定自噬抑制剂调节 TME，同时提供临床前原理，以启动下一代临床试验， 自噬抑制剂在ICI联合方案中比HCQ更有效和特异。我们的临床试验将提供 有价值的安全性和临床活性数据，这些数据也将指导开发更有效和特异性的 自噬抑制剂。

Project Summary – Project 2 A new approach that can overcome resistance to immune checkpoint inhibition (ICI) is a major unmet need for Stage IV melanoma patients. The overall goal of this project is to determine if combined ICI with autophagy inhibition can address this unmet need. Autophagy is a key resistance mechanism to chemotherapy and targeted therapy. More recently our work and the work of others has implicated autophagy as a resistance mechanism to immunotherapy. This raises a number of questions about which is the best approach to target autophagy and in which cell types is it most critical to target autophagy within the tumor microenvironment. This project will leverage deep collaborations with emerging biotechnology companies that have developed next generation chemical lysosomal and non-lysosomal autophagy inhibitors that are headed to the clinic. The following aims will test our overall hypothesis that lysosomal autophagy inhibition results in focused cellular pathway perturbations in cancer cells and immune cells that enhance the efficacy of ICI: Specific Aim 1 will determine the mechanism by which novel clinical grade autophagy inhibitors modulate tumor-immune interactions during ICI, focusing on effects on myeloid, tumor, T cell, and other immune cell phenotypes. We will compare the ability of each of these inhibitors to augment combined anti-PD-1 and anti CTLA-4 Ab in clinically relevant mouse models. We will focus on PPT1, a lysosomal thioesterase that regulates autophagy, and the major molecular target of chloroquine derivatives, and an exciting new target for cancer drug development. We will utilize our novel conditional Ppt1 KO mouse model, to compare the effects of ICI combined with Ppt1 KO in tumor cells, dendritic cells, and myeloid cells, and compare genetic inhibition to chemical Ppt1 inhibition on melanoma tumor growth. In Specific Aim 2 will determine changes in immunoprofiles of the TME in patients and preclinical models treated with combined ICI and autophagy inhibition. We will conduct the LIMIT melanoma trial, an adaptive phase I/II trial of nivolumab + HCQ and nivolumab + ipilimumab + HCQ in Stage IV melanoma patients. Novel PET imaging technologies will be used to track CD8+ T-cells in tumors and correlate CD8+ signal with histological CD8+ analysis and clinical response. In related pre-clinical mouse studies, we will use unbiased approaches to achieve a comprehensive view of changes in melanoma tumors treated with ICI and HCQ or ICI and DC661 during early response and resistance. Impact: Our study will identify the mechanism by which autophagy inhibitors modulate the TME while providing the preclinical rationale for launching next generation clinical trials using novel autophagy inhibitors more potent and specific than HCQ in ICI combination regimens. Our clinical trial will provide valuable safety and clinical activity data that will also guide the development of more potent and specific autophagy inhibitors.