The ontogeny and function of CD8 T cells in lung cancer

项目摘要/摘要 以免疫系统为靶点利用免疫疗法摧毁癌细胞已迅速成为一种很有前途的方法 该药物用于治疗癌症，并在部分患者中产生了强烈的临床反应。虽然首字母是 成功的案例提供了免疫系统可以被用来治疗癌症的概念的证据， 大多数患者没有获得持久的甚至是最初的好处，这突显了更好地了解 成功利用免疫系统消灭癌细胞的障碍。这项提议的重点是非 小细胞肺癌(NSCLC)，只有20%-40%的患者对 到目前为止，基于免疫的疗法。目前改善肺癌免疫疗法的一个主要障碍是缺乏 了解肿瘤发生过程中主要的T细胞反应。事实上，很少有研究能够 在生理时间框架内纵向解剖自然肺中CD8 T细胞的反应 微环境。以前的工作已经在肿瘤中发现了一组功能较弱的T细胞 表达转录因子T细胞因子1(TCF-1)，正是这个群体被认为是中介 高效的抗肿瘤反应。然而，如何在缺少这些细胞时生成这些细胞， 这些细胞在肿瘤发生过程中的发育轨迹，以及如何最好地针对这些细胞进行治疗 目的仍不清楚。该项目的目标是解决围绕TCF-1+的这些关键问题 用自体小鼠肺腺癌模型制备抗肿瘤CD8T细胞亚群 概述了人类非小细胞肺癌的时间尺度和解剖进展。利用尖端技术 方法和技术，包括单细胞RNA测序、共生和邻近标记，我们将 定义个体发育、分化、功能和决定应答抗肿瘤药物命运的决定因素 Tcf-1+CD8T细胞。在目标1中，我们将以我们的初步数据为基础，展示TCF内的异质性-- 通过阐明抗肿瘤CD8 T细胞的功能和转录状态来确定1+CD8 T细胞亚群 人口。我们将把我们在目标1中产生的转录数据与现有的人类单细胞RNA进行比较 测序以确定我们在模型中识别的CD8 T细胞状态的相关性。在《目标2》中我们将 使用邻近标记系统确定肿瘤-T细胞相互作用在T细胞表型和命运中的作用 识别最近与肿瘤细胞相互作用的CD8T细胞。在目标3中，我们将定义 淋巴结和肺TCF-1+CD8 T细胞，并将尝试在治疗上利用抗肿瘤CD8 T细胞 在DLN内种植更多的TCF-1+CD8 T细胞到肿瘤中。总而言之，我们的目标是 重新定义我们对抗肿瘤CD8 T细胞反应的理解，并有可能找到新的治疗方法 治疗癌症的途径。

PROJECT SUMMARY/ABSTRACT Targeting the immune system to destroy cancer cells using immunotherapy has rapidly emerged as a promising avenue for treating cancer, and has resulted in robust clinical responses in a subset of patients. While the initial success stories have provided proof of concept that the immune system can be harnessed to treat cancer, the majority of patients do not achieve long lasting or even initial benefit, highlighting the need to better understand the barriers to successfully using the immune system to eliminate cancer cells. The focus of this proposal is non- small cell lung cancer (NSCLC), which has only shown 20-40% of patients having an objective response to immune based therapies to date. A major obstacle to improving current immunotherapies in lung cancer is a lack of understanding of the overarching T cell response during tumorigenesis. Indeed, few studies have been able to longitudinally dissect the CD8 T cell response in a physiological time frame in the native lung microenvironment. Previous work has identified a population of less dysfunctional T cells in the tumor that express the transcription factor T cell factor 1 (TCF-1), and it is this population that is thought to mediate productive anti-tumor responses. However, how to generate these cells when they are lacking, the developmental trajectories of these cells during tumorigenesis, and how to best target these cells for therapeutic purposes remain unclear. The goal of this project is to address these critical questions surrounding the TCF-1+ subset of anti-tumor CD8 T cells using an autochthonous mouse model of lung adenocarcinoma that recapitulates both the time scale and anatomical progression of human NSCLC. Utilizing cutting edge methodologies and techniques, including single cell RNA sequencing, parabiosis and proximity labeling, we will define the ontogeny, differentiation, function, and determinants that dictate the fate of responding anti-tumor TCF-1+ CD8 T cells. In Aim 1, we will build on our preliminary data demonstrating heterogeneity within the TCF- 1+ CD8 T cell compartment by elucidating functionality and transcriptional status of anti-tumor CD8 T cell populations. We will compare the transcriptomic data we generate in Aim 1 to existing human single cell RNA sequencing to determine the relevance of the CD8 T cell states we identify in our model. In Aim 2 we will determine the role of tumor-T cell interactions in T cell phenotype and fate using a proximity labeling system to identify CD8 T cells that have recently interacted with tumor cells. In Aim 3, we will define the relationship of lymph node and lung TCF-1+ CD8 T cells, and will attempt to therapeutically harness anti-tumor CD8 T cells within the dLN to seed more TCF-1+ CD8 T cells into the tumor. Taken together, our aims are positioned to redefine our understanding of the anti-tumor CD8 T cell response and potentially identify new therapeutic avenues to treat cancer.