Determinants of response to cancer immunotherapy

项目摘要/摘要 虽然免疫疗法正在改变癌症治疗，但大多数患者并没有达到持久的效果。 回应。我们一直在研究对不同免疫检查点抑制剂的反应和耐药性，并 现在准备提出机制研究，旨在提供对免疫状态和 调节或抑制免疫检查点阻断反应的通路。使用高维无偏 单细胞rna-seq(ScRNAseq)，我们可以识别规范和非规范免疫效应器 介导抗肿瘤反应。我们认为非规范的效应器，如细胞毒性的CD4T细胞，我们 最近描述的，并不是我们目前的治疗方法有效地触发的。使用相同的单细胞 方法，我们可以在癌症患者中识别已知和新的细胞类型，这些细胞类型可以介导免疫 压制。在我们的第一个目标中，我们将确定包括药物在内的联合免疫疗法(S) 靶向特定的免疫抑制细胞可以增强这些细胞毒的CD4+T细胞的功能。通过 利用新辅助临床试验，患者在手术前接受免疫治疗，我们将使用单一 细胞基因组学和蛋白质组学，以确定这些组合是否可以1)针对所需的 免疫抑制机制，以及2)增强切除的规范和/或非规范的效应器 肿瘤。我们还将使用此方法来确定是否可以将这些特定的单元状态映射到 循环隔间。第二个目标是基于对我们集团的长期兴趣来定义 抗原特异性反应的动力学。使用单细胞T细胞受体测序，我们可以识别扩增的 T细胞克隆以及它们的定位。除了了解免疫疗法如何结合 诱导和调节肿瘤内特定的T细胞克隆型，我们可以确定免疫疗法如何 将功能可塑性诱导到想要或不想要的状态。第三个目标建立在我们20年的经验基础上 使用小鼠模型剖析免疫治疗反应或抵抗的潜在机制。我们会 利用耗竭和基因敲除确定非规范免疫效应器的功能意义 战略。我们还将确定联合免疫疗法如何产生有效或无效的抗- 肿瘤免疫反应，从而指导未来临床试验的设计。总而言之，我们的建议是基于 关于假说驱动的床到床和床到床的力学研究，目的是推进 癌症免疫疗法。凭借我们在该领域的深厚专业知识，领导专注于多学科团队的经验 在翻译研究方面，以及由基础科学和临床合作者组成的丰富网络；我们是独一无二的 有能力成功完成本提案中概述的研究计划。

PROJECT SUMMARY/ABSTRACT While immunotherapy is transforming cancer treatment, the majority of patients do not achieve durable responses. We have been studying response and resistance to different immune checkpoint inhibitors and are now poised to propose mechanistic studies aimed at providing an understanding of the immune states and pathways that mediate or inhibit response to immune checkpoint blockade. Using high-dimensional unbiased single-cell RNA-seq (scRNAseq), we can identify both canonical and non-canonical immune effectors that can mediate anti-tumor responses. We believe that non-canonical effectors such as cytotoxic CD4 T cells, which we have recently described, are not effectively triggered by our current treatments. Using the same single-cell approaches, we can identify both known and novel cell types in cancer patients that can mediate immune suppression. In our first objective, we will determine whether combination immunotherapies that include drug(s) targeting specific immunosuppressive cells can enhance the function of these cytotoxic CD4+ T cells. By leveraging neoadjuvant clinical trials where patients receive immunotherapy prior to surgery, we will use single cell genomics and proteomics to define whether these combinations can 1) target the desired immunosuppressive mechanisms, and 2) enhance canonical and/or non-canonical effectors within the resected tumors. We will also use this approach to determine whether we can map these specific cell states into the circulating compartment. The second objective is based on a longstanding interest in our group to define the dynamics of antigen-specific responses. Using single-cell T cell receptor sequencing, we can identify expanded T cell clones as well as follow their localization. In addition to understanding how immunotherapy combinations induce and modulate specific T cell clonotypes within the tumor, we can determine how immunotherapies can induce functional plasticity to desired or undesired states. The third objective builds on our 20 year experience using mouse models to dissect mechanisms underlying response or resistance to immunotherapy. We will determine the functional significance of non-canonical immune effectors using depletion and knock-out strategies. We will also determine how combination immunotherapies can elicit both effective or ineffective anti- tumor immune responses, thereby guiding the design of future clinical trials. In conclusion, our proposal is based on hypothesis-driven bench-to-bedside and bedside-to-bench mechanistic studies with the goal of advancing cancer immunotherapy. With our deep expertise in this field, experience leading multi-disciplinary teams focused on translational research, and a rich network of basic science and clinical collaborators; we are uniquely positioned to succeed in the research plan outlined in this proposal.