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

基本信息

批准号：
10536641
负责人：
Anand Kornepati
金额：
$ 4.04万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
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：
2020-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10536641
关键词：
Address Affect Antibodies Appearance Applications Grants Autophagocytosis Backcrossings Biometry Candidate Disease Gene Cell Differentiation process Cells Chemoresistance Color Combination immunotherapy Complement 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 Infiltration Interferons Knock-in Knowledge Lymphocyte Malignant Neoplasms Mediating Mediator Modeling Mus Mutation Natural Killer Cells Oncogenes Oncogenic Pathology Pathway interactions Prevention strategy Proliferating Property Protein Analysis Proto-Oncogenes Publishing Reporting Risk Role Signal Pathway Signal Transduction Sunburn Surface T-Lymphocyte Tamoxifen Testing Time Transfection Transplantation Tumor Burden Tumor Immunity UV induced UV induced DNA damage Western Blotting anti-PD-L1 cancer cell cancer immunotherapy cancer prevention cancer risk carcinogenesis cellular imaging clinically relevant confocal imaging driver mutation experimental study immune cell infiltrate 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 programmed cell death protein 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

项目摘要

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、病理学和小鼠模型来解决肿瘤内在 PD-L1 信号对黑色素瘤生成的影响。 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 细胞我们实验室的范式转变工作现在已经确定了从阴性肿瘤表面表达的 PD-L1 中分离出来的重要性。免疫致病性肿瘤细胞固有的 PD-L1 信号尚未报道。我们已发表的关于主要肿瘤细胞内在影响的初步数据。该拨款提案中阐述的肿瘤 PD-L1 表明这些信号可能有助于致癌。我们开发了一种新型小鼠黑色素瘤模型，其中PD-L1可以在起源癌细胞中被删除（黑色素细胞）由癌基因诱导（Nras/Cdkn2a）或癌基因诱导加诱导。紫外线照射造成的 DNA 损伤我们将使用这种独特的临床相关模型来测试我们的总体方案。假设黑色素细胞固有的 PD-L1 信号通过增强 Nras/Cdkn2a 和/或 DNA 损伤的致癌作用我们通过以下目标来解决这一假设：具体目标 1：定义 DNA 损伤反应 (DDR) 的细胞内在 PD-L1 控制及其后果对肿瘤微环境（TME）的影响我们将通过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紫外线照射造成的 DNA 损伤。细胞内在的 PD-L1 调节的 DNA 修复改变和随后的 TME 效应将使用以下方法进行评估 Westerns、共焦成像、RNA-seq、免疫分析、组织学和 DNA 损伤研究。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）我们还将治疗早期黑色素瘤发生过程中的基质和免疫微环境。使用 αPD-L1 或 DDR 激酶抑制剂的黑色素瘤小鼠并评估肿瘤内在 PD-L1 信号传导对肿瘤（例如突变负担）和微环境（免疫分析）的影响。具体目标 2：检验 PD-L1 促进癌基因启动和/或紫外线诱导的假设通过细胞内在效应区分黑色素瘤发生与免疫依赖性 PD-。 L1 信号对致癌作用的影响，将在小鼠中诱发黑色素瘤 ± 免疫细胞耗竭小鼠实验将与体外人体细胞研究相补充。将Nras、Cdkn2a等候选基因转染至正常人黑素细胞中，测试其效果肿瘤发生和 PD-L1 表达。