The Determinants of CD8 T cell Dysfunction in Oral Cavity Squamous Cell Carcinoma

口腔鳞状细胞癌 CD8 T 细胞功能障碍的决定因素

• 批准号: 10677361
• 负责人: Riley Dalton Zale Mullins
• 金额: $ 3.44万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677361
• 关键词: 3-Dimensional; Advisory Committees; Antibodies; Antigens; Antitumor Response; Binding Sites; CD8-Positive T-Lymphocytes; CRISPR interference; Cancer Control; Cancer Etiology; Cancer cell line; Cell Death; Cell Line; Cell Separation; Cells; ChIP-seq; Chimeric Proteins; Chronic; Clinical; Coculture Techniques; Collagen; DNA; DNA Binding; Data; Data Set; Development; Electroporation; Epithelium; Exclusion; Fellowship; Flow Cytometry; Functional disorder; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Goals; Head and Neck Cancer; Hybrids; Immune; Immunology; Immunotherapy; In Vitro; Infiltration; Inflammatory; Knowledge; Libraries; Literature; Malignant Neoplasms; Maps; Measures; Membrane; Mentorship; Mesenchymal; Methods; Modeling; Morbidity - disease rate; Organoids; Outcome; Paracrine Communication; Patients; Peripheral; Phenotype; Physicians; Proliferating; Protein Secretion; Proteins; RNA; Receptor Activation; Recurrence; Research; Resistance; Role; Sampling; Scientist; Signal Transduction; Smoking; Survival Rate; System; T cell infiltration; T cell response; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Therapeutic; Thymocyte Selection; Tissue-Specific Gene Expression; Training; Transposase; Tumor-Derived; Work; anti-PD-1; cancer cell; cancer therapy; cancer type; cell type; checkpoint inhibition; cytokine; cytotoxicity; exhaust; exhaustion; experimental study; functional restoration; hybrid protein; immune checkpoint; immunocytochemistry; improved; insight; knock-down; malignant mouth neoplasm; mortality; mouth squamous cell carcinoma; new therapeutic target; novel; progenitor; prognostic; protein expression; response; single-cell RNA sequencing; stem; therapeutic target; transcription factor; transcriptome sequencing; treatment strategy; tumor; vector control

PROJECT SUMMARY Head and neck cancer, including oral cavity squamous cell carcinoma (OCSCC), is the sixth leading cause of cancer and a major cause of morbidity and mortality. Unfortunately, the five-year survival rate of OCSCC has only slightly improved in the past 30 years. Recently, immune checkpoint inhibition (ICI) therapies have emerged as a promising treatment to improve OCSCC outcomes, yet only 14-22% of OCSCC patients respond to ICI. Thus, delineating the mechanisms of ICI resistance represents an opportunity to identify novel targets that may increase ICI efficacy and improve OCSCC patient survival. CD8 T cells are the primary immune cell type that kills malignant cells during ICI, and their dysfunction may contribute to ICI resistance. During the chronic response to a tumor, CD8 T cells enter an exhausted cell state – a dysfunctional phenotype characterized by decreased ability to lyse target cells. Expression of the transcription factor thymocyte selection-associated HMG box (TOX) is critical for inducing CD8 T cell exhaustion and is enriched in the exhausted CD8 T cell subset within single cell RNA-seq datasets of OCSCC patient tumor samples. The genes that are directly transcriptionally regulated by TOX that promote CD8 T cell dysfunction and may be therapeutically targeted, however, remain poorly defined. In addition to TOX, malignant cell signals may contribute to CD8 T cell dysfunction and ICI resistance. We discovered a hybrid epithelial/mesenchymal (HEM) malignant cell state in OCSCC that localizes to the tumor edge adjacent to CD8 T cells and is associated with exhaustion of these adjacent CD8 T cells. The mechanisms of HEM cells underlying this association are not well understood. The primary hypothesis of this proposal is that CD8 T cell dysfunction in OCSCC is driven by the direct gene targets of TOX in CD8 T cells and by immunosuppressive contact and paracrine signaling from HEM cells. The goal of this proposal is to define mechanisms of CD8 T cell dysfunction that may be targeted to improve ICI through the following Aims: In Aim 1, the direct gene targets of TOX and their role in CD8 T cell dysfunction will be defined in vitro using CD8 T cells isolated from OCSCC samples. In Aim 2, patient-derived CD8 T cells will be co-cultured with patient-matched OCSCC tumor-derived organoids or cancer cell lines expressing a model antigen. In these co-cultures, HEM- specific membrane and secreted proteins will be knocked-down in the malignant cells to determine their role in suppressing CD8 T cells. The proposed research will provide mechanistic insight into CD8 T cell dysfunction in OCSCC, thereby supporting development of new strategies to therapeutically activate CD8 T cells and improve ICI and OCSCC outcomes. Importantly, during this fellowship, I will pursue scientific and clinical activities under the direct mentorship of a comprehensive fellowship advisory committee composed of physician-scientists and experts in cancer and immunology research. My proposed training will be vital to accomplishing my goal of investigating cancer-immune interactions and maturing as an independent, academic physician-scientist.

项目摘要 头颈癌，包括口腔鳞状细胞癌（OCSCC），是第六个主要原因 癌症和发病率和死亡率的主要原因。不幸的是，OCSCC的五年生存率 在过去的30年中，只有略有改善。最近，出现了免疫检查点抑制（ICI）疗法 作为改善OCSCC结果的有前途的治疗方法，但只有14-22％的OCSCC患者对ICI做出了反应。 这是描述ICI耐药机制的机会，这是一个机会，可以识别可能 提高ICI效率并提高OCSCC患者的生存。 CD8 T细胞是主要的免疫细胞类型 在ICI期间杀死恶性细胞，其功能障碍可能导致ICI耐药性。在慢性期间 对肿瘤的反应，CD8 T细胞进入疲惫的细胞状态 - 功能障碍的表型，其特征是 提高了裂解靶细胞的能力。转录因子胸腺细胞选择相关HMG的表达 盒子（TOX）对于诱导的CD8 T细胞耗尽至关重要，并且富含耗尽的CD8 T细胞子集中 OCSCC患者肿瘤样品的单细胞RNA-seq数据集。直接转录的基因 由促进CD8 T细胞功能障碍的TOX调节，但可能会被热靶向 定义不佳。除毒To外，恶性细胞信号可能导致CD8 T细胞功能障碍和ICI 反抗。我们在OCSCC中发现了一个本地化的混合上皮/间质（HEM）恶性细胞状态 与CD8 T细胞相邻的肿瘤边缘，与这些相邻CD8 T细胞的精疲力尽有关。这 该关联的下摆细胞机制尚不清楚。主要假设 建议是OCSCC中的CD8 T细胞功能障碍是由CD8 T细胞中的TOX的直接基因驱动的 通过免疫抑制接触和来自HEM细胞的旁分泌信号传导。该提议的目的是定义 CD8 T细胞功能障碍的机制可能针对通过以下目的改善ICI的目标：在AIM 1中， TOX的直接基因靶标及其在CD8 T细胞功能障碍中的作用将在体外使用CD8 T细胞定义 从OCSCC样品中分离出来。在AIM 2中，患者衍生的CD8 T细胞将与患者匹配共同培养 OCSCC肿瘤衍生的类器官或表达模型抗原的癌细胞系。在这些共培养中，下摆 特定的膜和分泌的蛋白质将在恶性细胞中被击倒，以确定它们在 抑制CD8 T细胞。拟议的研究将为CD8 T细胞功能障碍提供机械洞察力 OCSCC，从而支持制定热激活CD8 T细胞并改善的新策略 ICI和OCSCC结果。重要的是，在此奖学金期间，我将在 由身体科学家和 癌症和免疫学研究专家。我建议的培训对于完成我的目标至关重要 研究癌症免疫相互作用，并成为独立的学术身体科学家。