Inflammatory cascades disrupt Treg function through epigenetic mechanisms

炎症级联反应通过表观遗传机制破坏 Treg 功能

• 批准号: 10555213
• 负责人: William A Faubion
• 金额: $ 49.85万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-10 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10555213
• 关键词: 3-Dimensional; Adoptive Cell Transfers; Adoptive Transfer; Anti-Inflammatory Agents; Architecture; Articulation; Automobile Driving; Binding; Bioinformatics; Biology; CCCTC-binding factor; CD4 Positive T Lymphocytes; Cell Therapy; Cell physiology; Cells; Chromatin; Chromatin Loop; Chromatin Structure; Chronic; Clinic; Complex; Crohn' s disease; DNA; Derivation procedure; Dermatologic; Development; Disease; Drug Targeting; ETS Family Protein; Enhancers; Environment; Epigenetic Process; Evaluation; Event; Exposure to; FOXP3 gene; Genes; Genetic Transcription; Goals; Grant; Human; Human Biology; Human Engineering; Immune; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Intestines; Investigational Therapies; Kinetics; Lesion; Lymphocyte; Maintenance; Malignant Neoplasms; Mediating; Mus; Neurologic; Pathogenicity; Pathway interactions; Patients; Peripheral; Phenocopy; Phenotype; Polycomb; Proteins; Public Health; Publishing; Qualifying; Regulatory T-Lymphocyte; Repression; Repressor Proteins; Research; Risk; Role; Shapes; Signal Transduction; Spleen; Spontaneous colitis; T-Lymphocyte; Testing; Therapeutic; Therapy trial; Work; cellular engineering; cytokine; epigenetic drug; epigenetic therapy; epigenome; epigenomics; experience; gene network; gene repression; gut inflammation; in vivo; inflammatory milieu; insight; laboratory development; novel; programs; promoter; prototype; therapeutic evaluation; therapeutic target

PROJECT SUMMARY/ABSTRACT CD4+FOXP3+lymphocytes are expanded within the intestinal inflammatory lesion of Crohn’s disease (CD); however, ongoing inflammation belies presumed anti-inflammatory function of this cell. FOXP3 is required for differentiation and function of T regulatory (TREG) cells. FOXP3-mediated gene repression is lost in intestinal T cells of CD patients. Indeed, “FOXP3+ Crohn’s cell (FOXP3+CD)” bears a transcriptional signal more closely related to the pro-inflammatory TH17 cell. Derivation, function, and therapeutic implications of the FOXP3+CD cell remain poorly understood. Our long-term goal is to dissect epigenetic mechanisms regulating TREG cellular differentiation and function in the setting of GI inflammation. Consequently, the objective is to identify mechanisms responsible for activation of the TH17 phenotypic transcriptional network within intestinal FOXP3+CD cells and test therapeutic opportunities to restore their regulatory function in vivo. Our central hypothesis is that a shared chromatin configuration between TREG and TH17 cells allows for rapid activation of a TH17-like pro-inflammatory gene program in intestinal FOXP3+TREG cells. A set of TH17-relevant genes are accessible, yet not active in human TREG cells. A defining feature of the TH17 program was significant enrichment of CCCTC binding factor (CTCF) motifs. CTCF, a well-known topologically-associated domain (TAD) insulator protein, also mediates intra-TAD chromosomal looping and enhancer-promoter interaction regulating gene transcription. Bioinformatic analysis of this TH17 program indicates an inhibitory role for Polycomb Repressor Complex 1 (PRC1). Deletion of PRC1 in murine FOXP3+ cells led to secretion of prototypic TH17-like cytokines in FOXP3+ cells, and spontaneous colitis. The rationale is that with mechanistic insight into the biology of intestinal TREG cell in inflammation, one can apply targeted FOXP3+ CD cell-directed therapy including engineering of human TREG cells for adoptive cell therapy trials. To test the central hypothesis and obtain the overall objective we will address the following three questions: Aim1: Why are CTCF motifs associated with the TH17 program? Aim2: Why are TH17-associated genes accessible yet not active in TREG cells? Aim3: Why do FOXP3+CD cells express prototypic TH17 cytokines? Upon conclusion, we will understand the epigenetic pathways and 3D chromatin architecture spawning the FOXP3+CD cell and mechanisms shaping their development and function. This contribution is significant as the FOXP3+CD cell is expanded in the Crohn’s lesion, phenocopies TH17 cell which is implicated in Crohn’s disease (and other inflammatory diseases), and represents an important epigenetic drug target for systemic or re-engineered cellular therapy. There is general acknowledgement of risk of FOXP3+ TREG cells converting to pathogenic FOXP3+TH17-like cells upon adoptive transfer in Crohn’s trials; yet, this project is the first to articulate both signature and precise epigenetic events that regulate activation of the TH17 program in TREG cells.

项目总结/摘要 CD 4 + FOXP 3+淋巴细胞在克罗恩病（CD）的肠道炎性病变内扩增; 然而，持续的炎症掩盖了该细胞的假定抗炎功能。需要FOXP 3 调节性T细胞（Treg）的分化和功能。FOXP 3介导的基因抑制在肠道中丢失 CD患者的T细胞。事实上，“FOXP 3+克罗恩氏细胞（FOXP 3 +CD）”携带更多的转录信号。 与促炎性TH 17细胞密切相关。的来源、功能和治疗意义 FOXP 3 +CD细胞仍然知之甚少。我们的长期目标是剖析表观遗传机制， TREG细胞分化和功能在胃肠道炎症的设置。因此，目标是 确定负责肠内TH 17表型转录网络激活的机制 FOXP 3 +CD细胞和测试治疗机会，以恢复其体内调节功能。我们的中央 一种假说是TREG和TH 17细胞之间共享的染色质构型允许快速活化 TH17-like proinflammatory gene program in intestinal FOXP3+TREG cells.一组TH 17相关基因 是可接近的，但在人Treg细胞中没有活性。TH 17项目的一个显著特点是 CCCTC结合因子（CTCF）基序的富集。CTCF，一个著名的拓扑关联域 (TAD)绝缘子蛋白也介导染色体内成环和增强子-启动子相互作用 调节基因转录。该TH 17程序的生物信息学分析表明， 多梳阻遏复合物1（PRC 1）。PRC 1在鼠FOXP 3+细胞中的缺失导致PRC 1的分泌。 FOXP 3+细胞中的原型TH 17样细胞因子和自发性结肠炎。基本原理是， 深入了解炎症中肠道Treg细胞的生物学，可以应用靶向FOXP 3 + CD细胞定向 治疗包括改造人TREG细胞用于过继性细胞治疗试验。为了测试中央 假设并获得总体目标，我们将解决以下三个问题：目标1：为什么CTCF 与TH 17项目相关的图案目标2：为什么TH 17相关基因在人类中可获得但不活跃？ Treg细胞？目的3：为什么FOXP 3 +CD细胞表达原型TH 17细胞因子？在结束时，我们将 了解产生FOXP 3 +CD细胞的表观遗传途径和3D染色质结构， 形成其发展和功能的机制。这种贡献是显著的，因为FOXP 3 +CD细胞是 在克罗恩病病变中扩增，表型模仿与克罗恩病有关的TH 17细胞（和其他 炎症性疾病），并代表了一个重要的表观遗传药物靶点， 细胞疗法人们普遍认识到FOXP 3 + Treg细胞转化为致病性Treg细胞的风险。 FOXP 3 + TH 17样细胞在克罗恩氏试验中过继转移;然而，这个项目是第一个阐明这两个 信号和精确的表观遗传事件，调节Treg细胞中TH 17程序的激活。