Estrogen-mediated immune regulation in human and experimental inflammatory bowel disease

人类和实验性炎症性肠病中雌激素介导的免疫调节

• 批准号: 10355534
• 负责人: Wendy Ann Goodman
• 金额: $ 45.76万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10355534
• 关键词: Address; Adoptive Transfer; Antigens; Area; Back; Biological Assay; Biological Response Modifiers; Biopsy; CD4 Positive T Lymphocytes; Candidate Disease Gene; Cell physiology; Cells; Chronic; Co-Immunoprecipitations; Colitis; Complex; Crohn' s disease; Data; Defect; Disease; Environment; Environmental Risk Factor; Equilibrium; Estradiol; Estrogen Receptor alpha; Estrogen Receptor beta; Estrogens; Event; Exhibits; FOXP3 gene; Female; Frequencies; Gene Expression Regulation; Genetic; Genetic Transcription; Goals; Gonadal Steroid Hormones; Homeostasis; Human; Ileitis; Immune; Immune Tolerance; Immune response; Immunoprecipitation; Immunosuppression; Impairment; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Intestinal Mucosa; Intestines; Knockout Mice; Knowledge; Lead; Mediating; Membrane; Modeling; Molecular; Mus; Nuclear Receptors; Pathogenesis; Pathway interactions; Patients; Peripheral; Phenotype; Publishing; Regulatory T-Lymphocyte; Resistance; Role; Sampling; Sequence Homology; Signal Transduction; T cell differentiation; T cell response; T-Lymphocyte; Technology; Testing; Tissues; Transfection; Transforming Growth Factor beta; Ulcerative Colitis; Work; chromatin immunoprecipitation; chronic inflammatory disease; experimental study; gut inflammation; immunoregulation; improved; in vivo; in vivo evaluation; insight; loss of function; microbial; murine colitis; novel; novel strategies; prevent; receptor; response; sex; transcriptional reprogramming; transcriptome sequencing

Project Summary/Abstract Regulatory T cell (Treg) immunosuppression is critical for maintaining immune tolerance to a diverse array of potential antigens in the intestinal mucosa. In patients with inflammatory bowel disease (IBD), chronic intestinal inflammation overwhelms local Treg function, allowing inflammation to persist. Our previous work and that of others have identified important roles for 17β-estradiol (estrogen, E2) signaling in promoting Treg differentiation and function. E2 signals through two nuclear receptors, alpha and beta (ERα, ERβ), to modulate gene transcription in target cells. Although they share high sequence homology, ERα and ERβ mediate distinct and often opposing functions on gene regulation. In previously published work, we showed that shifting the balance of E2 signaling towards ERα is generally pro-inflammatory, whereas shifting towards ERβ is generally protective. In recent preliminary studies using IBD patient samples, we observed significantly diminished ERβ expression in intestinal biopsy tissues and peripheral T cells from females with active Crohn’s disease (CD). We also found that ERβ-deficient T cells are resistant to ex vivo, TGFβ-dependent Treg differentiation, and that deletion of ERβ in a spontaneous ileitis model (SAMP/YitFC, “SAMP” mice) results in significant impairment of Treg transcriptional and functional responses, contributing to exacerbated inflammation. Therefore, the goal of this project is to determine the molecular and cellular mechanism(s) by which altered E2 signaling impacts Treg differentiation and function, contributing to intestinal inflammation. The mechanisms by which E2 signaling cross-talks with inflammatory signals to influence immune cell function are poorly understood. This proposal seeks to address this knowledge gap through our central hypothesis that dysregulated E2 signaling contributes to Treg transcriptional remodeling and loss-of-function, facilitating sustained intestinal inflammation in IBD. In Aim 1, we propose to delineate the molecular mechanisms by which ERα- and ERβ cross-talk with signaling downstream of TGFβ in primary T cells, influencing TGFβ-dependent Foxp3 expression and function. Aim 2 will determine the functional impact of rebalancing Treg-specific E2 signaling on intestinal inflammation in vivo, testing our hypothesis that augmenting Treg-specific ERβ signaling may prevent and/or rescue intestinal inflammation. Experiments will include adoptive transfer of ERβ-expressing Tregs to SAMP mice, as well as in vivo assays using a T cell-dependent colitis model. In Aim 3, we plan to determine the transcriptional and functional effects of rebalancing E2 signaling in CD patient Tregs using novel MaxCyte transfection technology, assessing ERα- and ERβ-specific effects on (i) TGFβ-dependent Foxp3 induction in naïve T cells and (ii) ex vivo suppressive function of Tregs. Successful completion of our proposed Aims will provide key mechanistic insight into the functional impact of E2 signaling in Tregs, an under-studied area with broad applicability to numerous diseases exhibiting dysregulation of ER expression and/or activation and subsequent reductions in Treg function.

项目摘要/摘要 调节性T细胞（TREG）免疫抑制对于维持对潜水员阵列的免疫耐受性至关重要 肠粘膜中的潜在抗原。在炎症性肠病（IBD）的患者中，慢性肠道 炎症淹没了局部Treg功能，使炎症持续存在。我们以前的工作和 其他人已经确定了17β-雌二醇（雌激素，E2）信号传导的重要作用 和功能。 E2通过两个核接收器Alpha和Beta（ERα，ERβ）信号，以调节基因 靶细胞中的转录。尽管它们具有高序列同源性，ERα和ERβ介导了不同的，并且 通常在基因调节方面具有相反的功能。在先前发表的工作中，我们证明了改变余额 E2向ERα的信号传导通常是促炎性的，而转移向ERβ的转移通常受到保护。 在最近使用IBD患者样本的初步研究中，我们观察到ERβ表达显着降低 在患有活跃的克罗恩病（CD）的女性的肠活检组织和周围T细胞中。我们还发现 ERβ缺陷型T细胞对离体有抵抗力，TGFβ依赖性Treg分化，并且缺失ERβ 在赞助肠胃炎模型（SAMP/YITFC，“ SAMP”小鼠）中导致Treg的重大损害 转录和功能反应，导致加剧的注射。因此，目标的目标 项目是为了确定E2信号影响改变的分子和细胞机制 Treg的分化和功能，导致肠炎。 E2的机制 与炎症信号的信号传导交叉对话，以影响免疫细胞功能。这 提案试图通过我们的中心假设来解决这一知识差距，即E2信号失调 有助于Treg转录重塑和功能丧失，支持持续的肠道 在AIM 1中，我们建议描述ERα-和ERβ的分子机制 与原代T细胞中TGFβ的信号传导的串扰，影响TGFβ依赖性FOXP3表达 和功能。 AIM 2将确定重新平衡Treg特异性E2信号对肠的功能影响 体内炎症，检验我们的假设，即增加Treg特异性ERβ信号传导可能会预防和/或 救援肠炎。实验将包括将ERβ表达Treg的自适应转移到SAMP 使用T细胞依赖性结肠炎模型的小鼠以及体内测定。在AIM 3中，我们计划确定 使用新型Maxcyte在CD患者Treg中重新平衡E2信号传导的转录和功能效应 转染技术，评估ERα-和ERβ特异性对（I）TGFβ依赖性FOXP3诱导的影响 幼稚的T细胞和（ii）Treg的体内抑制功能。成功完成我们提议的目标 将提供对E2信号在Tregs的功能影响的关键机械洞察力，这是一个不足的研究 对众多疾病的广泛适用性展示ER表达和/或的区域 Treg功能的激活和随后的降低。