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.

抽象的该F31多样性提案使用我们的新癌症研究了PDL1在早期黑色素瘤进展中的作用产卵细胞自动摄取小鼠模型。我们的总体假设是肿瘤内膜PDL1信号通过免疫和非免疫机制促进早期肿瘤进展。我们的建议将定义新颖的细胞中性PDL1信号在黑色素瘤细胞中，并建立了黑色素瘤，提供了第一个在没有潜在混淆的情况下，区分善意的生物肿瘤肿瘤内部PDL1信号传导先前PDL1复发细胞的基因PDL1KO引起的机制。我们将解决这个假设以以下目的： AIM 1检验了黑色素内细胞INTRINSIC PDL1信号促进黑色素瘤进展的假设和治疗敏感性。我们将研究黑素细胞 - 内膜PDL1信号，下游效应子和使用PDL1KO TNQ61R小鼠模型促进紫外线暴露促进早期肿瘤进展的贡献创建的和同窝材料控件，其中PDL1在黑素细胞中专门删除。转录组（RNA-seq）随着pdl1-null良性良性的进展，将评估蛋白质组学（Luminex，RPPA）动力学 NEVI至恶性黑色素瘤定义何时PDL1在致癌过程中出现并确定遗传和蛋白质组学的统治pdl1在体内和体外及其后果。 PDL1的调节致癌信号（例如NRA，BRAF，MEK，ERK，MTORC1）将使用小分子抑制剂和用于WT NRAS或BRAFV600E突变体NRASQ61R的体外CRISPR替换。 PDL1抑制免疫原性皮肤紫外线暴露后的刺激信号将在并行探索，并且免疫原性机制为定义，将在AIM 2中测试它们对从头肿瘤生长和治疗敏感性的贡献。 AIM 2定义了黑色素细胞内pdl1驱动进展和治疗脆弱性中的TME因子。我们将使用我们的PDL1KO TNQ61R模型来测试黑素细胞PDL1依赖性TME免疫动力学。随着时间的流逝，非恶性皮肤和诱导肿瘤诱导的肿瘤，随着肿瘤的流动而进行免疫种群细胞仪。免疫印迹，Luminex和QRT-PCR将测试免疫原性信号和趋化因子与肿瘤共定位的生产离体和免疫细胞将通过共聚焦成像评估。选择将研究已知会影响治疗功效的免疫原性途径（例如，凋亡，rig-i/mavs）类似的时尚。 PDL1KO TNQ61R小鼠的研究和具有ICB，免疫细胞耗竭和的细胞系研究免疫缺陷小鼠测试PDL1驱动的免疫依赖性靶向治疗脆弱性。免疫 PDL1/NRAS交叉讲座的结果将定义为AIM 1中的信号研究。