The ontogeny and function of CD8 T cells in lung cancer

项目概要/摘要 靶向免疫系统以使用免疫疗法摧毁癌细胞已迅速成为一种有前途的治疗方法。 这是一种治疗癌症的新途径，并在一部分患者中产生了稳健的临床反应。虽然最初的 成功的故事提供了概念证明，免疫系统可以用来治疗癌症， 大多数患者没有获得持久的甚至是初步的益处，这突出表明需要更好地了解 成功利用免疫系统消除癌细胞的障碍。这一建议的重点是非- 小细胞肺癌（NSCLC），其仅显示20-40%的患者具有对化疗的客观反应。 免疫疗法迄今为止改善目前肺癌免疫治疗的一个主要障碍是缺乏 了解肿瘤发生过程中T细胞的总体反应。事实上，很少有研究能够 纵向剖析天然肺中生理时间范围内的CD 8 T细胞应答 微环境以前的工作已经确定了肿瘤中功能障碍较少的T细胞群， 表达转录因子T细胞因子1（TCF-1），并且认为正是这个群体介导了 有效的抗肿瘤反应。然而，如何产生这些细胞时，他们缺乏， 这些细胞在肿瘤发生过程中的发育轨迹，以及如何最好地靶向这些细胞进行治疗， 目的仍然不清楚。本项目的目标是解决围绕TCF-1+的这些关键问题 抗肿瘤CD 8 T细胞亚群使用肺腺癌的本地小鼠模型， 概括了人类NSCLC的时间尺度和解剖学进展。利用切削刃 方法和技术，包括单细胞RNA测序，共生和邻近标记，我们将 定义个体发生、分化、功能和决定应答性抗肿瘤药物命运的决定因素 TCF-1+ CD 8 T细胞。在目标1中，我们将以初步数据为基础，证明TCF内的异质性- 通过阐明抗肿瘤CD 8 T细胞的功能性和转录状态的1+ CD 8 T细胞区室 人口。我们将比较我们在Aim 1中生成的转录组数据与现有的人类单细胞RNA 测序以确定我们在模型中鉴定的CD 8 T细胞状态的相关性。在目标2中， 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， 鉴定最近与肿瘤细胞相互作用的CD 8 T细胞。在目标3中，我们将定义 淋巴结和肺TCF-1+ CD 8 T细胞，并将尝试在治疗上利用抗肿瘤CD 8 T细胞 在dLN内接种更多的TCF-1+ CD 8 T细胞进入肿瘤。总的来说，我们的目标是 重新定义我们对抗肿瘤CD 8 T细胞应答的理解，并可能发现新的治疗方法。 治疗癌症的方法

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.