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.

项目摘要 在恶性肿瘤中，CD8+ T细胞可以识别并消除肿瘤细胞，但通常无法治愈疾病 由于免疫抑制肿瘤中的慢性激活导致了它们的抗肿瘤功能的逐渐丧失 微环境。响应健康组织中的感染，T细胞分化为组织居民记忆 细胞（TRM），在清除抗原后，可以在组织中暴露于调查并提供保护 重新感染。当在癌症患者肿瘤中发现TRM样T细胞时，对免疫疗法的反应得到了改善 并观察到更好的患者预后。但是，是否像TRM一样代表疲惫的“祖细胞” TIL，或者是耗尽光谱以外的单独的细胞状态，尚不清楚。 为了更好地理解TRM样TIL，疲惫状态和TRM记忆单元之间的关系，我们 直接比较了急性病毒感染的TRM，并从肿瘤中耗尽了TIL，以发现转录差异 在这些不同的T细胞状态之间。专注于TRM高度表达的基因，可以调节其 在组织中提高功能，我们询问当T细胞终止时，其中哪些被下调 精疲力尽，与功能丧失一致。这种方法确定了许多与蛋白质有关的基因 调节，包括多个特征不足的E3泛素连接酶。泛素调节蛋白质 蛋白质体系统是稳态的重要生物学过程，对于细胞分化和功能至关重要， 以前已证明对记忆T细胞身份很重要。与我们的初步数据相关， 我们发现疲惫的til在其细胞质中有过量的蛋白质，当我们强制执行时 在肿瘤特异性T细胞中鉴定出的E3泛素连接酶的表达，它可以更好地控制肿瘤和 改善了小鼠的存活率。因此，我们建议探索蛋白质稳态与 TRM，精疲力尽和类似TRM的TIL细胞命运。 AIM 1试图了解确定的E3泛素连接酶是否 通过使用癌症模型强制表达或淘汰这些癌症，影响疲惫或TRM细胞的命运 连接酶和研究精疲力尽的T细胞命运，或在急性病毒感染中强制表达或淘汰连接酶 并研究TRM细胞命运。 AIM 2试图通过IN识别这些E3泛素连接酶的蛋白质相互作用靶标 体外生物形象接近标记测定法和质谱法，然后验证体内目标，并确定是否是否 这些连接酶在TIL中的表达可以减少展开的蛋白质丰度。了解关系 在T细胞耗尽和TRM之间可以为这两个分化T的生物学提供重要的新见解 细胞种群并告知操纵T细胞命运对TRM样TIL的努力以使癌症受益 免疫疗法。