Determining the role of AIRE and AIRE-expressing tumor associated macrophages in tumor growth and immunity

确定 AIRE 和表达 AIRE 的肿瘤相关巨噬细胞在肿瘤生长和免疫中的作用

基本信息

批准号：
10596378
负责人：
Sophia Guldberg
金额：
$ 3.97万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
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：
2023-03-18 至 2026-03-17
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10596378
关键词：
ATAC-seq Antibodies Antigens Antitumor Response Autoimmune Autoimmune Diseases Autoimmunity Automobile Driving Biological Assay Biology Bone Marrow CD4 Positive T Lymphocytes California Cancer Etiology Cell surface Cells Cellular biology Chromatin Chromosome Mapping Coculture Techniques Cytometry Data Data Set Development Environment Epigenetic Process Equilibrium Fellowship Flow Cytometry Frequencies Funding Gene Expression Generations Genetic Genetic Transcription Goals Health Hematopoietic Human ITGAM gene Immune Immune response Immunology Immunotherapy In Vitro Institution Investigation Knock-out Laboratories MC38 Macrophage Malignant Neoplasms Mentorship Methods Modeling Mus Mutation Myeloid Cells Pathway interactions Peripheral Phenotype Play Polyglandular Autoimmune Syndrome Type I Population Regulation Regulator Genes Research Research Project Grants Role San Francisco Signal Pathway Solid Neoplasm Stimulant Syndrome System TNFRSF5 gene TNFSF11 gene TNFSF5 gene Testing Therapeutic Thymic epithelial cell Thymus Gland Tissues Training Tumor Immunity Tumor-associated macrophages Universities Work anti-CTLA4 anti-PD-1 anti-tumor immune response autoreactive T cell cancer immunotherapy cancer therapy career cell type central tolerance epigenomics gene function gene induction human RNA sequencing immune activation immune checkpoint blockade immunoregulation immunosuppressive macrophages improved in vivo knockout gene melanoma mouse model neoplastic cell novel novel therapeutics patient response peripheral tolerance prevent programs research and development response secondary lymphoid organ single cell analysis single-cell RNA sequencing skills synergism systemic autoimmunity therapy resistant tool transcriptome sequencing tumor tumor growth tumor progression

项目摘要

PROJECT SUMMARY/ABSTRACT AutoImmune Regulator gene (AIRE) prevents autoimmunity by promoting thymic deletion of self-reactive T cells. While most studied in the thymus, AIRE is also expressed in secondary lymphoid organs, where it is thought to contribute to peripheral tolerance through interaction with and deletion of self-reactive CD4+ T cells. However, AIRE’s role in the periphery is likely more multi-faceted than in the thymus. We recently identified AIRE expression in tumor associated macrophages (TAMs), which are known to inhibit anti-tumor immune responses. While Aire-expressing tumor associated macrophages (aTAMs) were only recently discovered by our group, prior work implicates AIRE in inhibiting anti-tumor immunity. Recently available tools to study AIRE in mice have facilitated investigation of peripheral AIRE expression and aTAMs. We have found aTAMs in several common solid tumor models expressing common macrophage markers (CD11b, F4/80, and CD64) by both flow cytometry and mass cytometry (CyTOF). Additionally, interrogation of several public human RNA-seq datasets supports AIRE expression in tumor resident myeloid cell types. This preliminary data together with the therapeutic potential for targeting aTAMs to improve cancer immunotherapy make this a population deserving of thorough functional investigation. This proposal will test the hypothesis that aTAMs are an immunosuppressive, pro-tumoral cell population. Aim 1 of this proposal will define the phenotypic, transcriptional, and epigenetic profiles of aTAMs. Aim 2 will determine the mechanism(s) driving induction of AIRE expression in aTAMs. Aim 3 will identify the role of aTAMs in regulating local and systemic immune responses during anti-tumor immunity. This research approach will be carried out using a variety of methods including single cell analysis via RNA-seq, CyTOF, flow cytometry, and ATAC-seq, ex vivo co-cultures, and in vivo assays utilizing novel genetic mouse models. These proposed studies will be the first characterization of AIRE expression in any tumor resident immune cells and will further establish our understanding of the function of peripheral AIRE- expressing cell types. This could result in the discovery of novel pathways relevant to therapeutic resistance and further our understanding of global AIRE function. Translationally, this work may identify novel perturbations to improve patient responses to immunotherapy. This research project and fellowship training will be conducted at a top-funded research institution, the University of California, San Francisco (UCSF), in the laboratories of Dr. Matthew Spitzer, Dr. Lewis Lanier, and Dr. James Gardner with expert mentorship from Dr. Matthew (Max) Krummel. Dr. Lanier has made extensive foundational discoveries in immune cell biology. Dr. Spitzer has expertise in systems immunology and single cell methods. Dr. James Gardner has expertise in the study of peripheral AIRE and mouse model generation. Dr. Max Krummel has expertise in tumor myeloid cell biology and cancer immunotherapies. Overall, this facility and team provide a rich training environment for completion of this research and development of professional skills necessary for a career in academic research.

项目概要/摘要自身免疫调节基因 (AIRE) 通过促进胸腺删除自身反应性 T 细胞来预防自身免疫。虽然大多数研究是在胸腺中进行的，但 AIRE 也在次级淋巴器官中表达，人们认为它在次级淋巴器官中表达通过与自身反应性 CD4+ T 细胞的相互作用和删除，有助于外周耐受。 AIRE 在外周的作用可能比在胸腺中的作用更加多方面。肿瘤相关巨噬细胞 (TAM) 中的表达，已知可抑制抗肿瘤免疫反应。虽然我们的团队最近才发现表达 Aire 的肿瘤相关巨噬细胞 (aTAM)，先前的工作表明 AIRE 可以抑制抗肿瘤免疫，最近可用的工具在小鼠中研究 AIRE。促进了外周 AIRE 表达和 aTAM 的研究，我们在几种常见的细胞中发现了 aTAM。通过流式细胞术表达常见巨噬细胞标志物（CD11b、F4/80 和 CD64）的实体瘤模型此外，对几个公共人类 RNA-seq 数据集的询问也支持了这一点。肿瘤驻留骨髓细胞类型中的 AIRE 表达。该初步数据与治疗数据一起。靶向 aTAM 来改善癌症免疫治疗的潜力使这一人群值得彻底研究该提案将检验 aTAM 是一种免疫抑制药物的假设。该提案的目标 1 将定义表型、转录和表观遗传。 aTAM 的概况将确定驱动 aTAM AIRE 表达的机制。图 3 将确定 aTAM 在抗肿瘤免疫过程中调节局部和全身免疫反应的作用。该研究方法将使用多种方法进行，包括通过 RNA-seq 进行单细胞分析， CyTOF、流式细胞术和 ATAC-seq、离体共培养以及利用新型基因小鼠的体内测定这些拟议的研究将是任何肿瘤中 AIRE 表达的第一个表征。常驻免疫细胞，并将进一步建立我们对外周 AIRE 功能的理解这可能会导致与治疗耐药性相关的新途径的发现。并进一步加深我们对全球 AIRE 功能的理解。从翻译角度来说，这项工作可能会发现新的扰动。该研究项目和奖学金培训将改善患者对免疫疗法的反应。在顶级资助的研究机构加州大学旧金山分校 (UCSF) 进行 Matthew Spitzer 博士、Lewis Lanier 博士和 James Gardner 博士的实验室，并得到 Dr. Lewis Lanier 博士的专家指导。 Matthew (Max) Krummel 博士在免疫细胞生物学领域取得了广泛的基础性发现。 Spitzer 拥有系统免疫学和单细胞方法方面的专业知识。 Max Krummel 博士拥有肿瘤骨髓细胞方面的专业知识，致力于外周 AIRE 和小鼠模型生成的研究。总体而言，该设施和团队为生物学和癌症免疫疗法提供了丰富的培训环境。完成这项研究并开发从事学术研究职业所需的专业技能。