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Mapping epigenetic regulatory circuits of Th17 pathogenicity in autoimmunedisease

绘制自身免疫性疾病中 Th17 致病性的表观遗传调控回路

基本信息

批准号：
9541948
负责人：
Pratiksha I Thakore
金额：
$ 5.9万
依托单位：
BROAD INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-03-01 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9541948
关键词：
Affect American Animal Model Anti-inflammatory Attenuated Autoimmune Diseases Autoimmune Responses Autoimmunity Binding Biological Assay Cell physiology Cells Cellular Assay Chromatin Chronic Computing Methodologies Data Data Set Development Disease Disease model Epigenetic Process Equilibrium Event Experimental Autoimmune Encephalomyelitis Foundations Functional disorder Gene Expression Gene Targeting Genes Genetic Genetic Transcription Helper-Inducer T-Lymphocyte Heterogeneity Homeostasis Immune In Vitro Individual Inflammation Inflammatory Inflammatory Bowel Diseases Inflammatory Response Interleukin-17 Intervention Lead Maps Measures Mediating Multiple Sclerosis Neuraxis Pathogenicity Phenotype Population Protective Agents Psoriasis Regulatory Element Regulatory T-Lymphocyte Rheumatoid Arthritis Role Supervision T-Lymphocyte Technology Testing Time Tissues Transposase Variant cytokine disorder control epigenetic variation epigenome epigenomics experimental study extracellular functional plasticity genome-wide in vitro Model in vitro activity in vivo in vivo Model insight mouse model pathogen prevent programs response single cell analysis single-cell RNA sequencing transcription factor

项目摘要

Immune cells have diverse functions, from regulatory roles in tissue homeostasis to inflammatory responses against foreign challenges. In autoimmune disease, affected immune cells acquire a pathogenic state characterized by chronic activation of proinflammatory genes. CD4+ Th17 cells exemplify this functional plasticity, acting as protective agents in gut function or harmful instigators of inflammation in autoimmune disease. Th17 cells have been shown to be disease-causing agents in several autoimmune disorders affecting >10 million Americans, including multiple sclerosis, inflammatory bowel disease, psoriasis, and rheumatoid arthritis. Previous studies have revealed considerable heterogeneity in the ability of Th17 cells to induce disease and become pathogenic in animal models of multiple sclerosis. My objective is to identify the epigenetic regulatory mechanisms that control Th17 plasticity toward a pathogenic fate in autoimmune disease. I hypothesize that Th17 effector function is encoded in the epigenome and this can be leveraged to identify transcription regulators and remodeling events that distinguish the pathogenic state from regulatory function in Th17 cells in autoimmune disease. By applying recent advances in massively parallel single-cell epigenomics, computational methods for predicting transcription factor binding and deciphering single-cell variation, and technologies for targeted genetic perturbation10,12,17, I will test my hypothesis through the following aims: (1) Use the assay for transposase accessible chromatin (ATAC)-seq to identify an epigenetic signature defining the Th17 pathogenic fate in the context of in vitro cytokine-induced Th17 polarization; (2) Characterize epigenetic heterogeneity in Th17 pathogenic cell fate using single-cell ATAC-seq during experimental autoimmune encephalomyelitis, an autoimmune disease model of multiple sclerosis, and use this heterogeneity to identify potential transcription factor regulators of Th17 autoimmunity; and (3) Perform genetic perturbation experiments during Th17 polarization and measure impact on gene expression and chromatin accessibility in order to determine mechanisms of candidate drivers for establishing the Th17 pathogenic fate. With these studies, I will reconstruct the epigenetic regulatory circuits, consisting of transcription factors, target genes, and regulatory elements that control the disease-causing, pro-inflammatory cell fate of Th17 cells. These studies will provide important insights into the epigenetic networks that control nuanced cell function and reveal targeted strategies to reprogram immune cell pathogenic states in autoimmune disease without affecting non-pathogenic Th17 function.

免疫细胞具有多种功能，从组织稳态的调节作用到炎症应对外来挑战。在自身免疫性疾病中，受影响的免疫细胞获得致病性的以促炎基因的慢性激活为特征的状态。CD 4 + Th 17细胞具有这种功能可塑性，作为肠道功能的保护剂或自身免疫性疾病中炎症的有害煽动者，疾病Th 17细胞已被证明是几种自身免疫性疾病中的致病因子， > 1000万美国人，包括多发性硬化症、炎症性肠病、牛皮癣和类风湿性关节炎关节炎先前的研究已经揭示了Th 17细胞诱导免疫应答的能力的相当大的异质性。疾病并在多发性硬化症的动物模型中变成致病性。我的目标是找出控制Th 17可塑性向自身免疫性疾病致病命运的表观遗传调节机制疾病我假设Th 17效应子功能是在表观基因组中编码的，这可以被利用来识别区分致病状态和调节状态的转录调节因子和重塑事件 Th 17细胞在自身免疫性疾病中的功能。通过应用大规模并行单细胞表观基因组学的最新进展，预测转录因子结合和破译单细胞变异，以及靶向遗传干扰10，12，17，我将通过以下目的来检验我的假设：（1）使用该测定法，转座酶可及染色质（ATAC）-seq以鉴定限定Th 17致病性的表观遗传标记（2）在体外苦参碱诱导的Th 17极化的背景下表征表观遗传异质性，在实验性自身免疫性脑脊髓炎中使用单细胞ATAC-seq检测Th 17致病细胞的命运，自身免疫性疾病模型的多发性硬化症，并使用这种异质性，以确定潜在的转录 Th 17自身免疫的因子调节剂;和（3）在Th 17期间进行遗传扰动实验极化和测量对基因表达和染色质可及性的影响，以确定用于建立Th 17致病命运的候选驱动因子的机制。通过这些研究，我将重建表观遗传调控回路，包括转录因子，靶基因和调控元件，控制致病，促炎细胞的命运， Th 17细胞这些研究将为控制细微细胞的表观遗传网络提供重要的见解功能和揭示有针对性的策略，以重新编程自身免疫性疾病中的免疫细胞致病状态而不影响非致病性Th 17功能。