Mechanisms of REST-mediated immunosuppression in cancer

REST 介导的癌症免疫抑制机制

基本信息

批准号：
10749289
负责人：
Daniel E Michaud
金额：
$ 6.91万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
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：
2023-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10749289
关键词：
Agitation Antigens Biological Assay Breast Cancer Cell CD8-Positive T-Lymphocytes CSF1R gene Cancer Etiology Cell secretion Cells Cellular Immunity Cessation of life Characteristics Coculture Techniques Cytometry Data Flow Cytometry Gene Expression Profile Genes Goals Grant Immune Immune response Immunofluorescence Immunologic Immunologist Immunosuppression Immunotherapeutic agent Immunotherapy In Vitro Infiltration Intrinsic factor KDR gene Knock-out Link Lymphangiogenesis Lymphatic Lymphatic Endothelium Lymphocyte Lymphocyte Activation Lymphocyte Function Lymphocyte Suppression Lymphocytic Infiltrate Macrophage Malignant Neoplasms Mammary Neoplasms Mediating Migration Assay Modeling Mus Myelogenous Neurosecretory Systems Outcome Pathway Analysis Patients Periodicity Phenotype Population Positioning Attribute Prognosis Proliferating RNA RNA analysis Refractory Regulation Research Research Proposals Resistance Rest Role Signal Transduction Solid Neoplasm T-Lymphocyte Technical Expertise Testing The Cancer Genome Atlas Therapeutic Training Transcription Repressor Tumor Promotion Tumor-Infiltrating Lymphocytes Tumor-associated macrophages United States Vascular Endothelial Growth Factors Woman Writing anti-PD-1 biomarker discovery biomarker identification cancer cell career chemokine cytokine cytotoxicity effector T cell functional loss immune cell infiltrate immune checkpoint blockade immunoregulation improved in vivo in vivo Model loss of function malignant breast neoplasm molecular subtypes nano-string novel prognostic of survival programmed cell death ligand 1 recruit response skills synergism targeted treatment transcriptome transcriptome sequencing treatment response treatment strategy tumor tumor growth tumor microenvironment tumor-immune system interactions

项目摘要

Project Abstract Tumor-intrinsic factors in cancer cells modulate the immune milieu to enable prolonged survival and growth of tumors. In breast cancer (BC), the 2nd leading cause of cancer-related deaths in women, molecular subtyping of these factors effectively guides targeted therapies, but does not guide immunotherapy approaches as immune infiltrates vary significantly within each subtype. The presence of tumor-infiltrating lymphocytes (TILs) is highly prognostic for survival and therapeutic benefit in BC, but the tumor-intrinsic factors governing their presence are not well defined. Using TCGA RNA analysis, I have identified loss of function in the transcriptional repressor REST as being a key correlate to reduced lymphocyte infiltration into tumors of multiple BC molecular subtypes. Loss of functional REST in BC (~15-20% of tumors) is associated with poorer prognoses, however, the mechanisms by which loss of REST function modulates the tumor immune microenvironment are not known and may provide novel targets for enhancing therapeutic responses. I recently generated a murine Rest knockout BC line (Rest-less) and my in vivo studies confirmed that Rest-less tumors contain significantly less lymphocytes while also revealing a significant increase in pro-tumor macrophages, a cell population our lab has identified as being key modulators of lymphocyte suppression in BC. Furthermore, my in vitro studies identified lymphangiogenic Vegfc and Vegfd among the most differentially upregulated genes in Rest-less BC cells. Tumor lymphangiogenesis is linked to T cell suppression in solid tumors and may provide a reason for Rest-mediated suppression of lymphocytes. Based on my preliminary data, I hypothesize that lymphocyte suppression in Rest- less tumors is orchestrated by the polarization of tumor-associated macrophages (TAMs) and lymphatic endothelium to a suppressive phenotype. I propose two specific aims to test my hypothesis. In Aim 1, I will determine the role of tumor intrinsic Rest on TAM-mediated lymphocyte suppression using co-culture assays of TAMs from Rest-less tumors together with lymphocytes in vitro and macrophage depletion through anti-CSF-1R treatment in an antigen-specific (GFP; JEDI) Rest-less model in vivo. In Aim 2, I will evaluate how Vegfr3, the receptor for Vegfc and Vegfd, regulates the REST-less tumor microenvironment and responsiveness to immunotherapy. To accomplish this task, I will test anti-Vegfr3 blockade on Rest-less tumors with or without anti- PD1 and comprehensively analyze tumor growth, lymphatic depletion, and immune responses using RNAseq, spectral cytometry, and cyclic immunofluorescence. Our proposed research will provide an understanding of a previously uncharacterized facet of REST-mediated immune suppression in BC and will use novel murine orthotopic models to test strategies that block immune suppressive mechanisms. Ultimately, I anticipate my findings will reveal targetable mechanism(s) to inhibit Rest-mediated immunosuppression in BC and provide a considerable impact on the treatment of REST-less tumors overall.

项目摘要癌细胞中的肿瘤内部因素调节免疫环境使得能够长期生存和生长肿瘤。在乳腺癌（BC）中，女性与癌症相关死亡的第二大原因，分子亚型这些因素有效地指导了针对疗法，但没有指导免疫疗法的方法为免疫在每个亚型中浸润均有显着差异。肿瘤浸润淋巴细胞（TILS）的存在高度在卑诗省的生存和治疗益处的预后，但是肿瘤内在的因素是定义不当。使用TCGA RNA分析，我已经确定了转录阻遏物中功能的丧失休息是与减少淋巴细胞浸润到多个BC分子亚型肿瘤中的关键相关的。卑诗省功能休息的损失（占肿瘤的约15-20％）与预后较差有关尚不清楚肿瘤免疫微环境的REST功能损失调节的机制，并且可以提供新的目标来增强治疗反应。我最近产生了一个小鼠休息淘汰赛 BC系（无休息）和我的体内研究证实，静止肿瘤的淋巴细胞明显较小同时，同时也揭示了亲肿瘤巨噬细胞的显着增加，我们的实验室已确定为是卑诗省淋巴细胞抑制的关键调节剂。此外，我的体外研究确定了在不休息的BC细胞中最差异上调的基因中的淋巴管植入VEGFC和VEGFD。瘤淋巴管生成与实体瘤的T细胞抑制有关，并可能为休息介导的理由提供抑制淋巴细胞。根据我的初步数据，我假设在休息中抑制淋巴细胞通过肿瘤相关巨噬细胞（TAM）和淋巴管的极化来策划较少的肿瘤内皮到抑制表型。我提出了两个特定的目的来检验我的假设。在AIM 1中，我会使用共培养测定法确定肿瘤固有休息在TAM介导的淋巴细胞抑制中的作用通过抗CSF-1R的体外和巨噬细胞的淋巴细胞从体外和巨噬细胞耗尽。在体内的抗原特异性（GFP; JEDI）的无休息模型中进行处理。在AIM 2中，我将评估Vegfr3如何 VEGFC和VEGFD的受体调节无休息的肿瘤微环境和对免疫疗法。为了完成这项任务，我将在有或没有抗的无休息肿瘤上测试抗VEGFR3封锁 PD1并全面分析了使用RNASEQ的肿瘤生长，淋巴耗竭和免疫反应，光谱细胞仪和环状免疫荧光。我们提出的研究将提供对以前在不列颠哥伦比亚省的静止介导的免疫抑制的方面未表征，将使用新型鼠原位模型测试阻止免疫抑制机制的策略。最终，我期待我的调查结果将揭示有针对性的机制，以抑制卑诗省的静止介导的免疫抑制并提供总体上对无休息的肿瘤的治疗产生了相当大的影响。