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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

基本信息

批准号：
10853530
负责人：
Wendy Ann Goodman
金额：
$ 3.14万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
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：
2023-09-01 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10853530
关键词：
Address Adoptive Transfer Antigens Area Backcrossings Biological Assay Biological Response Modifiers Biopsy CD4 Positive T Lymphocytes Candidate Disease Gene Cell Separation Cell physiology Cells Chronic Co-Immunoprecipitations Colitis Complex Crohn&apos 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 Immunosuppression Impairment Inflammation Inflammatory Inflammatory Bowel Diseases Intestinal Mucosa Intestines Knockout Mice Knowledge Mediating Membrane Modeling Molecular Mus Nuclear Receptors Pathogenesis Pathway interactions Patients Peripheral Phenotype Predisposition 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 Western Blotting Work chromatin immunoprecipitation chronic inflammatory disease experimental study gut inflammation immunoregulation improved in vivo insight loss of function microbial murine colitis novel novel strategies prevent receptor response sex transcriptional reprogramming transcriptome sequencing

项目摘要

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.

项目概要/摘要调节性 T 细胞 (Treg) 免疫抑制对于维持对多种不同病原体的免疫耐受至关重要肠粘膜中的潜在抗原。对于炎症性肠病 (IBD) 患者，慢性肠道炎症会压倒局部 Treg 功能，导致炎症持续存在。我们之前的工作以及其他人已经确定 17β-雌二醇（雌激素，E2）信号在促进 Treg 分化中的重要作用和功能。 E2 通过两个核受体 α 和 β（ERα、ERβ）发出信号来调节基因靶细胞中的转录。尽管它们具有高度的序列同源性，但 ERα 和 ERβ 介导不同且通常对基因调控的功能相反。在之前发表的作品中，我们证明了改变平衡 E2 信号转向 ERα 通常是促炎性的，而转向 ERβ 通常是保护性的。在最近使用 IBD 患者样本的初步研究中，我们观察到 ERβ 表达显着减少存在于患有活动性克罗恩病 (CD) 的女性的肠道活检组织和外周 T 细胞中。我们还发现 ERβ 缺陷型 T 细胞对离体、TGFβ 依赖性 Treg 分化有抵抗力，并且 ERβ 缺失在自发性回肠炎模型（SAMP/YitFC，“SAMP”小鼠）中，Treg 显着受损转录和功能反应，导致炎症加剧。因此，本次活动的目标该项目旨在确定改变 E2 信号传导影响的分子和细胞机制 Treg 的分化和功能，导致肠道炎症。 E2的机制人们对信号与炎症信号的交叉影响免疫细胞功能知之甚少。这该提案旨在通过我们的中心假设（E2 信号失调）来解决这一知识差距有助于 Treg 转录重塑和功能丧失，促进持续的肠道 IBD 中的炎症。在目标 1 中，我们建议描述 ERα- 和 ERβ 的分子机制与原代 T 细胞中 TGFβ 信号下游信号串扰，影响 TGFβ 依赖性 Foxp3 表达和功能。目标 2 将确定重新平衡 Treg 特异性 E2 信号传导对肠道的功能影响体内炎症，检验我们的假设，即增强 Treg 特异性 ERβ 信号传导可以预防和/或拯救肠道炎症。实验将包括将表达 ERβ 的 Tregs 过继转移至 SAMP 小鼠，以及使用 T 细胞依赖性结肠炎模型的体内测定。在目标 3 中，我们计划确定使用新型 MaxCyte 重新平衡 CD 患者 Tregs 中 E2 信号传导的转录和功能效应转染技术，评估 ERα 和 ERβ 对 (i) TGFβ 依赖性 Foxp3 诱导的特异性影响幼稚 T 细胞和 (ii) Tregs 的离体抑制功能。成功完成我们提出的目标将为Tregs中E2信号传导的功能影响提供关键的机制见解，Tregs是一种尚未充分研究的细胞该领域对许多表现出 ER 表达失调和/或 Treg 功能的激活和随后的减少。