Analysis of the E7-Mediated Mechanism of MHC Class I Repression in HNSCC

E7 介导的 HNSCC I 类 MHC 抑制机制分析

基本信息

批准号：
10066535
负责人：
Elizabeth Gensterblum-Miller
金额：
$ 3.85万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10066535
关键词：
3-Dimensional Acetylation Address Alanine Antitumor Response Architecture Autoantigens Binding Binding Sites Biological Assay CD3 Antigens CD8-Positive T-Lymphocytes CD8B1 gene CRISPR/Cas technology Cell Line Cells Cessation of life ChIP-seq Chromatin Chromatin Structure Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Cytotoxic T-Lymphocytes DNA Data Detection Disease Etiology Flow Cytometry Gene Expression Regulation Genes Genetic Genetic Transcription Goals HLA-A gene HPV-High Risk Head and Neck Squamous Cell Carcinoma Human Papillomavirus Human papillomavirus 16 Human papillomavirus 18 Immune Immune Evasion Immunotherapy Incidence Interferon Type II Knock-out MHC Class I Genes Major Histocompatibility Complex Malignant Neoplasms Mass Spectrum Analysis Mediating Modeling Molecular Oncogenic Oncoproteins Organoids Pathogenicity Pathway interactions Patients Peripheral Blood Mononuclear Cell Pharmacologic Substance Production Proteins RNA Polymerase II Regulation Regulatory Pathway Repression Scanning Site Stains Surface T-Cell Activation T-Lymphocyte Testing Therapeutic Transcriptional Regulation Transfection United States Validation Work anti-tumor immune response base cancer type genome-wide genomic locus human old age (65+)improved inhibitor/antagonist knockout gene malignant oropharynx neoplasm neoantigens neoplastic cell new therapeutic target novel novel therapeutic intervention overexpression prevent

项目摘要

Abstract Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer type in the world, and is responsible for over 8,000 deaths in the United States each year. The number of HNSCC cases caused by human papilloma virus (HPV) is on the rise, especially in patients 40-65 years old. HPV is currently associated with 80% of oropharyngeal cancers and 5-10% of HNSCC cases from other sub-sites. High-risk HPV strains, most commonly HPV16 and HPV18, cause the preponderance of HPV-positive HNSCC. Current data suggests that in HNSCC, HPV16 limits immune detection by preventing surface presentation of the major histocompatibility complex class I (MHC-I). In HPV-positive HNSCC, MHC-I surface presentation level is associated with the cytotoxic T cell-mediated anti-tumor immune response and thus the identification of strategies to prevent HPV-mediated MHC-I repression may have a significant therapeutic benefit for patients receiving immunotherapies. There some data that suggest E7 negatively regulates transcription of MHC-I constituent genes; however, the detailed mechanism of E7-mediated MHC-I repression is poorly characterized. Here, I have generated two E7-overexpressing HPV-negative HNSCC cell lines and confirmed that E7 protein downregulates transcription of MHC-I. We have developed systematic and logical approaches including CRISPR/CAS9 profiling that we propose to leverage to characterize the molecular mechanism(s) by which E7- represses MHC-I in these models, and HPV+ HNSCC models. Further, we have developed a 3D organoid-based co-culture assay in which these models are co-cultured with patient-matched peripheral blood mononuclear cells (PBMCs) to test the functional effects of de-repression of MHC-I expression. My central hypothesis is that identification of the molecular mechanisms by which HPV16_E7 diminish the expression of MHC-I will lead to the advancement of therapeutic strategies that enhance tumor cell recognition by activated T- cells. I will address this hypothesis through the following aims: 1) Detail the molecular mechanism(s) of MHC locus repression by HPV16_E7 in HNSCC, and 2) Qualify HPV16_E7-dependent MHC class I regulatory pathways in HNSCC. My long-term goal is to develop new therapeutic approaches that improve the overall survival of HPV+ patients, and in doing so, I hope to characterize the specific mechanisms by which HPV16_E7 can prevent MHC-I surface expression in HPV-positive HNSCC.

抽象的头颈鳞状细胞癌 (HNSCC) 是世界上第六种最常见的癌症类型，美国每年造成 8,000 多人死亡。引起的 HNSCC 病例数人乳头瘤病毒 (HPV) 呈上升趋势，特别是在 40-65 岁的患者中。目前与 HPV 相关 80% 的口咽癌和 5-10% 的 HNSCC 病例来自其他亚站点。高危 HPV 毒株，最常见的是 HPV16 和 HPV18，导致 HPV 阳性 HNSCC 占多数。目前的数据表明在 HNSCC 中，HPV16 通过阻止主要抗原的表面呈现来限制免疫检测 I 类组织相容性复合体 (MHC-I)。在 HPV 阳性 HNSCC 中，MHC-I 表面表达水平为与细胞毒性 T 细胞介导的抗肿瘤免疫反应相关，从而鉴定预防 HPV 介导的 MHC-I 抑制的策略可能对患者有显着的治疗益处接受免疫疗法。有一些数据表明 E7 负向调节 MHC-I 的转录组成基因；然而，E7 介导的 MHC-I 抑制的详细机制尚不清楚。在这里，我生成了两种 E7 过表达 HPV 阴性 HNSCC 细胞系，并证实 E7 蛋白下调 MHC-I 的转录。我们开发了系统性和逻辑性的方法，包括我们建议利用 CRISPR/CAS9 分析来表征 E7-的分子机制在这些模型和 HPV+ HNSCC 模型中抑制 MHC-I。此外，我们还开发了一种基于 3D 类器官的共培养测定，其中这些模型与患者匹配的外周血单核细胞共培养 (PBMC) 来测试 MHC-I 表达去抑制的功能效果。我的中心假设是 HPV16_E7 减少 MHC-I 表达的分子机制的鉴定将导致治疗策略的进步，通过激活的 T-增强肿瘤细胞识别细胞。我将通过以下目标来阐述这一假设：1) 详细说明 MHC 的分子机制 HNSCC 中 HPV16_E7 的位点抑制，以及 2) 验证 HPV16_E7 依赖的 MHC I 类监管 HNSCC 中的通路。我的长期目标是开发新的治疗方法来改善整体 HPV+ 患者的存活率，在此过程中，我希望能够描述 HPV16_E7 的具体机制可以阻止 HPV 阳性 HNSCC 中 MHC-I 表面表达。