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

• 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
• 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

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 依赖性 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。