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: 9911802
• 负责人: Anand Kornepati
• 金额: $ 3.44万
• 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/9911802
• 关键词: Address; Affect; Antibodies; Appearance; Applications Grants; Autophagocytosis; Backcrossings; Biometry; Candidate Disease Gene; Cell Differentiation process; Cells; 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/antagonist; insight; kinase inhibitor; melanocyte; melanoma; metaplastic cell transformation; mouse model; neoplasm immunotherapy; neoplastic cell; next generation; novel; programmed cell death ligand 1; response; 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.

该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，以解决肿瘤内在PD-L1信号对黑色素瘤形成的影响。的 αPD-L1免疫疗法的主要作用机制被认为是保护PD-1+抗肿瘤T细胞 来自阴性肿瘤表面表达的PD-L1。我们实验室的范式转变工作现在已经定义了重要的 免疫病理肿瘤细胞-内在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。 起源有助于致癌作用。我们发表的和初步的数据，主要肿瘤细胞的内在影响， 肿瘤PD-L1，在这个拨款提案中阐明，表明这些信号可能有助于致癌。 我们开发了一种新的小鼠黑色素瘤模型，其中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/Cdkn 2a）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 黑素细胞内源性PD-L1信号通过增强PD-L1的表达促进黑素瘤发生的假说。 Nras/Cdkn 2a的致癌作用和/或DNA损伤。我们在以下目标中处理这一假设： 具体目标1：定义DNA损伤反应（DDR）的细胞内在PD-L1控制及其后果 肿瘤微环境（TME）我们将通过以下方式在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上UV照射引起的DNA损伤。肿瘤 将使用以下方法评估细胞内在PD-L1调节的DNA修复改变和随后的TME效应 Western、共聚焦成像、RNA-seq、免疫分析、组织学和DNA损伤研究。我们的新模型 将显示黑素细胞PD-L1的出现如何改变DDR和其他致癌介质（例如，mTOR）造型 间质和免疫微环境随时间的变化。我们还将治疗 用αPD-L1或DDR激酶抑制剂治疗荷黑色素瘤小鼠，并评估肿瘤内源性PD-L1信号传导 对肿瘤的后果（例如，突变负荷）和微环境（免疫分析）。 具体目的2：检验PD-L1促进癌基因启动和/或UV诱导的假设 黑色素瘤通过细胞内在的影响。为了区分细胞内源性与免疫依赖性PD- L1信号传导对致癌作用的影响，将在小鼠中诱导黑色素瘤±免疫细胞耗竭 有特异性抗体。小鼠实验将与体外人细胞研究相补充， 将Nras、Cdkn 2a和其他候选基因转染到正常人黑素细胞中，以测试对黑素细胞的影响。 肿瘤发生和PD-L1表达。