Role of cell-intrinsic effects of PD-L1 in carcinogenesis

PD-L1 的细胞内在效应在致癌过程中的作用

• 批准号: 10311517
• 负责人: Anand Kornepati
• 金额: $ 3.56万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10311517
• 关键词: Address; Affect; Antibodies; Appearance; Applications Grants; Autophagocytosis; Backcrossings; Biometry; Candidate Disease Gene; Cell Differentiation process; Cells; Chemoresistance; Color; Combination immunotherapy; Complement; Cyclin-Dependent Kinase Inhibitor 2A; Cytometry; DNA Damage; DNA Repair; DNA damage checkpoint; Data; Event; Experimental Neoplasms; FDA approved; FRAP1 gene; Flow Cytometry; Generations; Genes; Growth; Histologic; Histology; Human; Immune; Immune Evasion; Immune checkpoint inhibitor; Immune system; Immunity; Immunotherapy; In Vitro; Interferons; Knock-in; Knowledge; Lymphocyte; Malignant Neoplasms; Mediating; Mediator of activation protein; Modeling; Mus; Mutation; Natural Killer Cells; Oncogenes; Oncogenic; Pathology; Pathway interactions; Prevention strategy; Property; Protein Analysis; Proto-Oncogenes; Publishing; Reporting; Risk; Role; Signal Pathway; Signal Transduction; Sunburn; Surface; T-Lymphocyte; Tamoxifen; Testing; Time; Transplantation; Tumor Burden; Tumor Immunity; Tumor-infiltrating immune cells; UV induced; UV induced DNA damage; Western Blotting; anti-PD-1; anti-PD-L1; cancer cell; cancer immunotherapy; cancer prevention; cancer risk; carcinogenesis; cellular imaging; clinically relevant; confocal imaging; driver mutation; experimental study; in vivo; inhibitor; insight; kinase inhibitor; melanocyte; melanoma; melanomagenesis; metaplastic cell transformation; mouse model; neoplasm immunotherapy; neoplastic cell; next generation; novel; programmed cell death ligand 1; response; risk prediction; shift work; therapy resistant; transcriptome sequencing; treatment response; treatment strategy; tumor; tumor immunology; tumor initiation; tumor microenvironment; tumor progression; tumor-immune system interactions; tumorigenesis; ultraviolet irradiation

This proposal assembles experts in tumor immunotherapy, tumor immunology, melanoma, biostatistics, pathology and mouse models to address effects of tumor intrinsic PD-L1 signals on melanomagenesis. The principal mechanism of action of αPD-L1 immunotherapy is thought to be protection of PD-1+ anti-tumor T cells from negative tumor surface-expressed PD-L1. Paradigm shifting work from our lab has now defined important immunopathogenic tumor cell-intrinsic PD-L1 signals. It is unreported whether PD-L1 in a given cancer cell of origin contributes to carcinogenesis. Our published and preliminary data on major tumor cell-intrinsic effects of tumor PD-L1, illuminated in this grant proposal, suggest that these signals could contribute to carcinogenesis. We developed a novel mouse melanoma model in which PD-L1 can be deleted in the cancer cell of origin (melanocyte). Melanomas are induced by either oncogene induction (Nras/Cdkn2a) or oncogene induction plus DNA damage from UV irradiation. We will use this unique and clinically relevant model to test our overarching hypothesis that melanocyte-intrinsic PD-L1 signals promote melanomagenesis by enhancing the oncogenic effects of Nras/Cdkn2a and/or DNA damage. We address this hypothesis in the following aims: Specific Aim 1: Define cell-intrinsic PD-L1 control of the DNA damage response (DDR) and consequent effects on the tumor microenvironment (TME). We will induce tumors in our mouse melanoma model via either tamoxifen-induced oncogene induction or oncogene induction plus DNA damage by UV irradiation. Tumor cell-intrinsic PD-L1-regulated alterations in DNA repair and subsequent TME effects will be assessed using Westerns, confocal imaging, RNA-seq, immune analyses, histologic and DNA damage studies. Our novel model will show how appearance of melanocyte PD-L1 alters DDR and other oncogenic mediators (e.g., mTOR) sculpt the stroma and immune microenvironment over time during early melanomagenesis. We will also treat melanoma-bearing mice with αPD-L1 or DDR kinase inhibitors and assess tumor intrinsic PD-L1-signaling consequence on tumor (e.g., mutational burden) and microenvironment (immune analyses). Specific Aim 2: Test the hypothesis that PD-L1 promotes initiation of oncogene and/or UV induced melanomagenesis through cell-intrinsic effects. To distinguish cell-intrinsic versus immune-dependent PD- L1 signaling consequences on carcinogenesis, melanomas will be induced in mice ± immune cell depletions with specific antibodies. Mouse experiments will be complemented with human cell studies in vitro by transfecting Nras, Cdkn2a and other candidate genes into normal human melanocytes to test effects on tumorigenesis and PD-L1 expression.

这项提案汇集了肿瘤免疫治疗、肿瘤免疫学、黑色素瘤、生物统计学、 病理学和小鼠模型研究肿瘤固有PD-L1信号对黑色素瘤发生的影响。这个 αPD-L1免疫治疗的主要作用机制被认为是保护PD-1+抗肿瘤T细胞 从肿瘤表面阴性表达的PD-L1。我们实验室的范式转换工作现在定义了重要的 免疫致病肿瘤细胞固有的PD-L1信号。目前还没有报道PD-L1在给定的癌细胞中是否存在 原产地有助于癌症的发生。我们已发表的关于主要肿瘤细胞的初步数据-内在效应 这项拨款提案中阐明的肿瘤PD-L1表明，这些信号可能有助于癌症的发生。 我们开发了一种新的小鼠黑色素瘤模型，在该模型中，PD-L1可以在起源的癌细胞中缺失 (黑素细胞)。黑色素瘤是由癌基因诱导(NRAS/CDKN2a)或癌基因诱导加 紫外线照射造成的DNA损伤。我们将使用这个独特的和临床相关的模型来测试我们的总体 黑素细胞固有PD-L1信号通过促进黑色素瘤发生的假说 NRAS/CDKN2a和/或DNA损伤的致癌作用。我们从以下几个方面阐述了这一假设： 具体目标1：确定DNA损伤反应(DDR)的细胞固有PD-L1控制及其结果 对肿瘤微环境(TME)的影响。我们将通过以下方式在我们的小鼠黑色素瘤模型中诱导肿瘤 他莫昔芬诱导的癌基因诱导或癌基因诱导加紫外线照射所致的DNA损伤。肿瘤 细胞固有的PD-L1调节的DNA修复变化和随后的TME效应将使用 西片、共聚焦成像、rna-seq、免疫分析、组织学和dna损伤研究。我们的新模式 将展示黑素细胞PD-L1的出现如何改变DDR和其他致癌介质(例如，mTOR)的造型 黑色素瘤发生早期随时间变化的间质和免疫微环境。我们也会请你 αPD-L1或DDR激酶抑制剂对黑色素瘤荷瘤小鼠的作用及肿瘤内源性PD-L1信号的评估 对肿瘤的影响(例如，突变负担)和微环境(免疫分析)。 特定目标2：验证PD-L1促进癌基因启动和/或紫外线诱导的假设 黑色素瘤的发生通过细胞的内在效应。为了区分细胞内源性和免疫依赖型帕金森病- L1信号对小鼠致癌、黑色素瘤的影响±免疫细胞耗竭 用特定的抗体。小鼠实验将与人体细胞体外研究相辅相成 将NRAS、CDKN2a等候选基因导入正常人黑素细胞，检测其对黑素细胞的影响 肿瘤发生与PD-L1的表达