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在因素调节免疫环境，使癌细胞能够延长生存和生长肿瘤。乳腺癌 (BC) 是导致女性癌症相关死亡的第二大原因，其中分子亚型这些因素可以有效指导靶向治疗，但不能像免疫治疗那样指导免疫治疗方法。每种亚型的浸润情况差异很大。肿瘤浸润淋巴细胞 (TIL) 的存在高度 BC 的生存和治疗益处的预后，但控制其存在的肿瘤内在因素是没有明确定义。使用 TCGA RNA 分析，我发现转录抑制子功能丧失 REST 是减少淋巴细胞浸润多种 BC 分子亚型肿瘤的关键相关因素。 BC 中功能性 REST 的丧失（约 15-20% 的肿瘤）与较差的预后相关，然而， REST 功能丧失调节肿瘤免疫微环境的机制尚不清楚，可能为增强治疗反应提供新的靶点。我最近生成了小鼠休息淘汰赛 BC 系（无休息）和我的体内研究证实，无休息肿瘤含有明显更少的淋巴细胞同时还揭示了促肿瘤巨噬细胞的显着增加，我们的实验室确定了一个细胞群是 BC 淋巴细胞抑制的关键调节剂。此外，我的体外研究发现淋巴管生成 Vegfc 和 Vegfd 属于 Rest-less BC 细胞中差异最大的上调基因。瘤淋巴管生成与实体瘤中的 T 细胞抑制有关，可能为休息介导的淋巴管生成提供了一个原因抑制淋巴细胞。根据我的初步数据，我假设休息时淋巴细胞抑制肿瘤相关巨噬细胞（TAM）和淋巴管的极化可以减少肿瘤的发生内皮细胞转变成抑制性表型。我提出了两个具体目标来检验我的假设。在目标 1 中，我将使用共培养测定确定肿瘤内在休息对 TAM 介导的淋巴细胞抑制的作用来自无休息肿瘤的 TAM 与体外淋巴细胞以及通过抗 CSF-1R 消除巨噬细胞体内抗原特异性（GFP；JEDI）无休息模型中的治疗。在目标 2 中，我将评估 Vegfr3（即 Vegfc 和 Vegfd 受体，调节无 REST 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-Rest-less肿瘤上测试抗-Vegfr3阻断作用 PD1 并使用 RNAseq 全面分析肿瘤生长、淋巴耗竭和免疫反应，光谱细胞术和循环免疫荧光。我们提出的研究将提供对 BC 中 REST 介导的免疫抑制的先前未表征的方面，并将使用新型小鼠原位模型用于测试阻断免疫抑制机制的策略。最终，我预计我的研究结果将揭示抑制 BC 中休息介导的免疫抑制的靶向机制，并提供整体上对无 REST 肿瘤的治疗产生了相当大的影响。