Ubiquitin ligase regulation of tissue-resident T cell and anti-tumor activity

泛素连接酶对组织驻留 T 细胞的调节和抗肿瘤活性

• 批准号: 10726015
• 负责人: Ananda W Goldrath
• 金额: $ 23.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10726015
• 关键词: Acute; Antigens; Biological; Biological Assay; Biological Process; Biology; Biotin; CD8-Positive T-Lymphocytes; CD8B1 gene; CRISPR/Cas technology; Cancer Model; Cancer Patient; Cancerous; Cell Differentiation process; Cell Surface Receptors; Cell physiology; Cells; Cellular biology; Characteristics; Chronic; Cytosol; Cytotoxic T-Lymphocytes; Data; Disease; Doxycycline; Enzymes; Epigenetic Process; Family; Genes; Genetic Transcription; Homeostasis; Immune; Immunotherapy; In Vitro; Infection; Inflammatory; Knock-out; Label; Ligase; Lymphocyte Function; Lymphocytic choriomeningitis virus; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Memory; Metabolic; Mus; Patient-Focused Outcomes; Patients; Play; Population; Process; Protein Engineering; Proteins; Regulation; Resistance; Role; Surveys; System; T cell differentiation; T memory cell; T-Lymphocyte; Therapeutic; Tissues; Tumor-Infiltrating Lymphocytes; Ubiquitin; Ubiquitination; Virus Diseases; cancer immunotherapy; cancer therapy; chimeric antigen receptor T cells; cytokine; cytotoxic; differential expression; engineered T cells; exhaust; exhaustion; high reward; high risk; immune checkpoint blockade; improved; in vivo; insight; loss of function; member; misfolded protein; mouse model; multicatalytic endopeptidase complex; neoplastic cell; progenitor; protein degradation; protein transport; proteostasis; receptor expression; response; stem cell differentiation; tissue resident memory T cell; transcription factor; tumor; tumor growth; tumor microenvironment; tumor-immune system interactions; ubiquitin ligase; ubiquitin-protein ligase

PROJECT SUMMARY In malignancies, CD8+ T cells can recognize and eliminate tumor cells, but often fail to cure disease due to their progressive loss of antitumor function resulting from chronic activation in the immunosuppressive tumor microenvironment. In response to infection in healthy tissues, T cells differentiate into tissue-resident memory cells (TRM), and after clearance of antigen can remain lodged in tissues to survey and provide protection from reinfection. When TRM-like T cells are found in cancer patient tumors, improved responses to immunotherapy and better patient outcomes are observed. However, whether TRM-like TIL represent ‘progenitors’ of exhausted TIL, or are a separate cell state outside the exhaustion spectrum is still unclear. To better understand the relationship among TRM-like TIL, exhaustion states, and TRM memory cells, we directly compared TRM from acute viral infection and exhausted TIL from tumors to find transcriptional differences between these distinct T cell states. Focusing on genes highly expressed by TRM that may mediate their enhanced functions in tissues, we asked which of those were downregulated as T cells became terminally exhausted, coincident with loss of function. This approach identified numerous genes related to protein regulation, including multiple under-characterized E3 ubiquitin ligases. Protein regulation by the ubiquitin proteosome system is an essential biological process for homeostasis, crucial for cell differentiation and function, and has been previously shown to be important for memory T cell identity. Correlated with our preliminary data, we found that exhausted TIL have an excess of unfolded proteins in their cytosol, and when we enforced expression of the identified E3 ubiquitin ligases in tumor-specific T cells, it allowed for better tumor control and improved mouse survival. Therefore, we propose to explore the relationship between protein homeostasis and TRM, exhaustion, and TRM-like TIL cell fates. Aim 1 seeks to understand if the identified E3 ubiquitin ligases influence exhaustion or TRM cell fate by using mouse models of cancer to enforce expression or knock out these ligases and study exhaustion T cell fate, or to enforce expression or knock out the ligases in acute viral infection and study TRM cell fate. Aim 2 seeks to identify the protein-interaction targets of these E3 ubiquitin ligases by in vitro BioID proximity labeling assay and mass spectrometry, then verify the targets in vivo TIL and determine if expression of these ligases in TIL can decrease unfolded protein abundance. Understanding the relationship between T cell exhaustion and TRM can provide vital new insights into the biology of these two differentiated T cell populations and inform efforts to manipulate T cell fates towards TRM-like TIL to benefit cancer immunotherapy.

项目概要 在恶性肿瘤中，CD8+ T细胞可以识别并消除肿瘤细胞，但往往无法治愈疾病，因为 由于免疫抑制肿瘤的慢性激活而导致其抗肿瘤功能逐渐丧失 微环境。为了响应健康组织的感染，T 细胞分化成组织驻留记忆 细胞（TRM），清除抗原后可以保留在组织中以进行调查并提供保护 再次感染。当在癌症患者肿瘤中发现 TRM 样 T 细胞时，对免疫治疗的反应会改善 并且观察到更好的患者结果。然而，类似 TRM 的 TIL 是否代表了疲惫的“祖先”？ TIL 或耗尽谱之外的独立细胞状态仍不清楚。 为了更好地理解类似 TRM 的 TIL、耗尽状态和 TRM 存储单元之间的关系，我们 直接比较急性病毒感染的 TRM 和肿瘤的耗尽 TIL 以发现转录差异 这些不同的 T 细胞状态之间。重点关注 TRM 高表达的基因，这些基因可能介导其 增强组织中的功能，我们想知道当 T 细胞终末期时，其中哪些功能被下调 精疲力竭，与功能丧失同时发生。这种方法鉴定了许多与蛋白质相关的基因 调节，包括多种未充分表征的 E3 泛素连接酶。泛素对蛋白质的调节 蛋白体系统是体内平衡的重要生物过程，对于细胞分化和功能至关重要， 之前已被证明对记忆 T 细胞身份很重要。与我们的初步数据相关， 我们发现耗尽的 TIL 在其胞质中含有过量的未折叠蛋白质，当我们强制执行时 已鉴定的 E3 泛素连接酶在肿瘤特异性 T 细胞中的表达，可以更好地控制肿瘤， 提高小鼠的存活率。因此，我们建议探索蛋白质稳态与 TRM、耗竭和类似 TRM 的 TIL 细胞命运。目标 1 旨在了解已识别的 E3 泛素连接酶是否 通过使用小鼠癌症模型强制表达或敲除这些细胞来影响衰竭或 TRM 细胞的命运 连接酶并研究耗竭 T 细胞的命运，或在急性病毒感染中强制表达或敲除连接酶 并研究 TRM 细胞命运。目标 2 旨在通过以下方式识别这些 E3 泛素连接酶的蛋白质相互作用靶标： 体外 BioID 邻近标记测定和质谱分析，然后验证体内 TIL 的目标并确定是否 这些连接酶在 TIL 中的表达可以降低未折叠蛋白的丰度。了解关系 T 细胞耗竭和 TRM 之间的关系可以为这两种分化 T 细胞的生物学提供重要的新见解 细胞群并为操纵 T 细胞命运向 TRM 样 TIL 的努力提供信息，以造福癌症 免疫疗法。