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The role of tumor cell-of-origin-specific PDL1 on tumorigenesis and tumor progression

肿瘤细胞源特异性PDL1在肿瘤发生和进展中的作用

基本信息

批准号：
10679453
负责人：
Carlos Ontiveros
金额：
$ 3.77万
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：
2023
资助国家：
美国
起止时间：
2023-08-03 至 2027-08-02
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10679453
关键词：
Acceleration Address Affect Autophagocytosis BRAF gene Benign Bioinformatics Biological Cell Line Cells Clustered Regularly Interspaced Short Palindromic Repeats Complement DDR1 gene DNA Damage Data Dependence Dose Exposure to FDA approved Flow Cytometry Generations Genetic Harvest Image Immune Immune Evasion Immunohistochemistry Impairment In Vitro Interferon Type I Interferons Investigation Knowledge MEKs Malignant - descriptor Malignant Neoplasms Malignant neoplasm of ovary Malignant neoplasm of urinary bladder Medical Melanoma Cell Mentors Modeling Mus Mutate Nevus Non-Malignant Oncogenic Outcome Pathologic Pathway interactions Population Prediction of Response to Therapy Production Proteomics Quantitative Reverse Transcriptase PCR Regulation Reporting Role Signal Pathway Signal Transduction Skin Stimulator of Interferon Genes Surface T-Cell Depletion Testing Time Training Translating Transplantation Treatment Efficacy Tumor Immunity Tumor Promotion Tumor Stem Cells Tumor-Derived Tumor-Infiltrating Lymphocytes UV Radiation Exposure Western Blotting Work cancer cell cancer imaging cancer immunotherapy carcinogenesis carcinogenicity career development chemokine clinical translation clinically relevant confocal imaging cytokine experimental study immune cell infiltrate immunogenic immunogenicity improved in vivo in vivo evaluation inhibitor insight melanocyte melanoma melanomagenesis mouse model mutant neoplastic cell novel novel therapeutics pharmacologic programmed cell death ligand 1 programmed cell death protein 1 response response biomarker skills small molecule small molecule inhibitor therapy resistant transcriptome sequencing transcriptomics treatment strategy tumor tumor growth tumor initiation tumor microenvironment tumor progression tumor-immune system interactions tumorigenesis

项目摘要

ABSTRACT This F31-diversity proposal investigates the role of PDL1 in early melanoma progression using our novel cancer cell-of-origin autochthonous mouse model. Our overarching hypothesis is that tumor-intrinsic PDL1 signals promote early tumor progression by both immune and non-immune mechanisms. Our proposal will define novel cell-intrinsic PDL1 signals in the melanoma cell-of-origin and established melanomas providing the first model distinguishing bona-fide biologic tumor-intrinsic PDL1 signaling in the absence of potentially confounding mechanisms due to genetic PDL1KO of a previously PDL1-replete tumor cell. We will address this hypothesis with the following aims: Aim 1 Test the hypothesis that melanocyte-intrinsic PDL1 signals promote early melanoma progression and treatment sensitivities. We will investigate melanocyte-intrinsic PDL1 signals, downstream effectors, and contributions of UV exposure facilitating early tumor progression using the PDL1KO TNQ61R mouse model we created, and littermate controls, in which PDL1 is specifically deleted in melanocytes. Transcriptomic (RNA-seq) and proteomic (Luminex, RPPA) dynamics will be assessed as melanocytes progress from PDL1-null benign nevi to malignant melanomas defining when PDL1 emerges during carcinogenesis and determining the genetic and proteomic milieu governing PDL1 expression in vivo and in vitro and its consequences. PDL1 regulation of oncogenic signals (e.g., NRAS, BRAF, MEK, ERK, mTORC1) will be tested using small molecule inhibitors and in vitro CRISPR replacement of mutant NrasQ61R for WT Nras or BRAFV600E. PDL1 suppression of immunogenic STING signals following skin UV exposure will be explored in parallel and as immunogenicity mechanisms are defined, their contributions to de novo tumor growth and treatment sensitivities will be tested in Aim 2. Aim 2 Define TME factors in melanocyte-intrinsic PDL1-drive progression and treatment vulnerabilities. We will test melanocyte PDL1-dependent TME immune dynamics using our PDL1KO TNQ61R model by harvesting non-malignant skin and induced tumor over time, assessing immune populations as tumors progress with flow cytometry. Immunoblots, Luminex, and qRT-PCR will test immunogenic STING signals and chemokine production ex vivo and immune cell co-localization with tumor will be assessed by confocal imaging. Alternative immunogenic pathways known to affect treatment efficacy (e.g., pyroptosis, RIG-I/MAVS) will be studied in similar fashion. Studies of PDL1KO TNQ61R mice and derived cell lines with ICB, immune cell depletion, and immune deficient mice test PDL1-driven immune-dependent targetable treatment vulnerabilities. Immune outcomes of PDL1/Nras cross talk will be defined complementary to signaling studies in Aim 1.

抽象的这项 F31 多样性提案利用我们的新型癌症研究了 PDL1 在早期黑色素瘤进展中的作用细胞源本地小鼠模型。我们的首要假设是肿瘤固有的 PDL1 信号通过免疫和非免疫机制促进早期肿瘤进展。我们的提案将定义黑色素瘤细胞起源和已建立的黑色素瘤中的新型细胞内在 PDL1 信号提供了第一个在没有潜在混淆的情况下区分真正的生物肿瘤内在 PDL1 信号传导的模型机制归因于先前充满 PDL1 的肿瘤细胞的遗传 PDL1KO。我们将解决这个假设目标如下：目标 1 检验黑色素细胞固有的 PDL1 信号促进早期黑色素瘤进展的假设和治疗敏感性。我们将研究黑素细胞固有的 PDL1 信号、下游效应器和使用 PDL1KO TNQ61R 小鼠模型，紫外线照射促进早期肿瘤进展的贡献和同窝对照，其中黑素细胞中的 PDL1 被特异性删除。转录组学（RNA-seq）当黑素细胞从 PDL1 缺失良性进展时，将评估蛋白质组学（Luminex、RPPA）动态痣到恶性黑色素瘤定义癌发生过程中 PDL1 何时出现并确定遗传体内和体外控制 PDL1 表达的蛋白质组环境及其后果。 PDL1 调节致癌信号（例如 NRAS、BRAF、MEK、ERK、mTORC1）将使用小分子抑制剂进行测试，体外 CRISPR 将突变体 NrasQ61R 替换为 WT Nras 或 BRAFV600E。 PDL1 免疫原性抑制将同时探索皮肤紫外线暴露后的 STING 信号，并且免疫原性机制正在研究中。根据定义，它们对肿瘤从头生长和治疗敏感性的贡献将在目标 2 中进行测试。目标 2 定义黑素细胞固有 PDL1 驱动进展和治疗脆弱性中的 TME 因素。我们将使用我们的 PDL1KO TNQ61R 模型通过采集来测试黑色素细胞 PDL1 依赖性 TME 免疫动力学随着时间的推移，非恶性皮肤和诱发肿瘤，随着肿瘤随血流进展而评估免疫群体细胞计数术。免疫印迹、Luminex 和 qRT-PCR 将测试免疫原性 STING 信号和趋化因子离体生产和免疫细胞与肿瘤的共定位将通过共聚焦成像进行评估。选择将研究已知影响治疗效果的免疫原性途径（例如细胞焦亡、RIG-I/MAVS）类似的时尚。对 PDL1KO TNQ61R 小鼠和衍生细胞系进行 ICB、免疫细胞耗竭和免疫缺陷小鼠测试 PDL1 驱动的免疫依赖性靶向治疗漏洞。免疫 PDL1/Nras 串扰的结果将被定义为对目标 1 中的信号研究的补充。