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.

抽象的头颈部鳞状细胞癌（HNSCC）是世界上第六种最常见的癌症类型，是每年在美国造成8,000多人死亡。由人乳头状瘤病毒（HPV）正在上升，尤其是在40-65岁的患者中。 HPV当前关联 80％的口咽癌和5-10％的HNSCC病例来自其他子站点。高风险的HPV菌株，最常见的是HPV16和HPV18，导致HPV阳性HNSCC的优势。当前数据建议在HNSCC中，HPV16通过防止表面表现出来限制了免疫检测组织相容性复杂I类（MHC-I）。在HPV阳性HNSCC中，MHC-I表面表现水平为与细胞毒性T细胞介导的抗肿瘤免疫反应相关，从而鉴定防止HPV介导的MHC-1抑制的策略可能对患者具有显着的治疗益处接受免疫疗法。一些数据表明E7负调节MHC-I的转录组成基因；但是，E7介导的MHC-1抑制的详细机制的特征很差。在这里，我产生了两个过表达的HPV阴性HNSCC细胞系，并确认E7蛋白下调MHC-I的转录。我们已经开发了系统和逻辑的方法，包括我们建议利用CRISPR/CAS9分析，以表征E7-的分子机制在这些模型中压抑MHC-I和HPV+ HNSCC模型。此外，我们已经开发了一个基于3D器官的共培养测定法与这些模型与患者匹配的外周血单核细胞共培养（PBMC）测试MHC-I表达抑制的功能效应。我的中心假设是识别HPV16_E7降低MHC-I表达的分子机制将导致治疗策略的发展，从而通过激活的T-增强肿瘤细胞识别细胞。我将通过以下目的解决这一假设：1）详细说明MHC的分子机制 HPV16_E7在HNSCC中抑制基因座和2）符合HPV16_E7依赖性MHC I类调节 HNSCC的途径。我的长期目标是开发新的治疗方法，以改善整体 HPV+患者的生存，在此过程中，我希望表征HPV16_E7的特定机制可以防止HPV阳性HNSCC中的MHC-I表面表达。