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Role of regulatory T cell glucocorticoid-induced leucine zipper (GILZ) in the chronically inflamed intestine

调节性 T 细胞糖皮质激素诱导的亮氨酸拉链 (GILZ) 在慢性炎症肠道中的作用

基本信息

批准号：
10215504
负责人：
Wendy Ann Goodman
金额：
$ 12.08万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-13 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10215504
关键词：
Adrenal Cortex Hormones Age Anti-Inflammatory Agents Antiinflammatory Effect Apoptosis Autoimmune Diseases Biological Assay C57BL/6 Mouse CD3 Antigens CD4 Positive T Lymphocytes Cell physiology Characteristics Chromium Chronic Colitis Crohn&apos s disease Cytokine Signaling Data Digestive System Disorders Dinitrobenzenes Disease Environment Epithelial Epithelial Cells Estradiol Estrogen Receptor alpha Estrogen Receptor beta Estrogens Exhibits FOXP3 gene Female Frequencies Future Gene Expression Profile Genes Genetic Transcription Genomics Glucocorticoid Receptor Glucocorticoids Goals Gonadal Steroid Hormones Human Ileitis Immunomodulators Immunosuppression Impairment Inflammation Inflammatory Inflammatory Bowel Diseases Interleukin-10 Intestinal Diseases Intestinal Mucosa Intestinal permeability Intestines Knockout Mice Lead Leucine Zippers Link MAP Kinase Gene Maintenance Mediating Messenger RNA Modeling Mucous Membrane Mus Nuclear Receptors Operative Surgical Procedures Pathology Pathway interactions Patients Peripheral Phenotype Phosphorylation Population Heterogeneity Proteins Receptor Signaling Regulation Regulatory T-Lymphocyte Repression Research Resistance Role Sampling Signal Transduction Spleen Sulfonic Acids System T cell differentiation T-Lymphocyte T-cell inflamed Testing Transcription Factor AP-1 Transcriptional Regulation Transfection Transforming Growth Factor beta Transgenic Mice Translating Up-Regulation Uterus autoinflammatory chronic inflammatory disease cohort design experimental study glucocorticoid receptor alpha hormonal signals improved in vivo inflammatory disease of the intestine insight intestinal homeostasis male next generation novel personalized medicine receptor binding single-cell RNA sequencing steroid hormone tool transcription factor transcriptome transcriptome sequencing

项目摘要

Project Summary/Abstract Glucocorticoid-induced leucine zipper (GILZ) is a broadly-expressed transcription factor whose expression is induced via endogenous or synthetic glucocorticoid (GC) signaling through the glucocorticoid receptor (GR). GILZ is thought to meditate many of GCs’ anti-inflammatory effects in T lymphocytes, such as induction of TGFβ signaling and inhibition of NFκB, MAPK, and AP-1 pathways. Conditional deletion of GILZ in CD4+ T cells leads to enhancement of experimental dinitrobenzene sulfonic acid (DNBS)-induced colitis, a Th1-mediated form of experimental inflammatory bowel disease (IBD). Steroid hormones, including corticosteroids and sex steroid hormones such as 17β-estradiol (estrogen, E2), demonstrate considerable promiscuity in receptor binding. E2 has been shown to antagonize GILZ in human uterine epithelial cells, but little is known regarding the mechanism(s) by which it may contribute to GILZ expression and/or function in T cells. Our preliminary data reveal a robust induction of GILZ at the mRNA and protein levels in regulatory T cells (Tregs) isolated from Crohn’s disease (CD) patients and SAMP/YitFC (“SAMP”) mice, a spontaneous model of Crohn’s-like disease. GILZ upregulation (“GILZhigh”) in these Tregs was associated with a relative decrease in estrogen receptor beta (ERβ) expression, suggesting that GILZ expression in Tregs may be responsive to estrogen signaling. Furthermore, GILZhigh Tregs exhibited reduced suppressive function compared to their GILZlow counterparts, suggesting that optimal GILZ-mediated immunosuppression in Tregs may require intact ERβ signaling. Several chronic inflammatory and autoimmune diseases exhibit reductions in ERβ expression and/or activity, leading to the intriguing possibility that diminished ERβ expression contributes to inflammation via disruption of normally-protective Treg GILZ-mediated mechanisms. Our hypothesis is that the protective functions of Treg- specific GILZ require intact ERβ signaling, and therefore fail to mediated sufficient immunoprotection in ERβ-depleted environments, such as the IBD intestine. The goal of this project is to determine the mechanism(s) by which reduced ERβ-specific signaling influences the expression and function of GILZ in IBD- associated Tregs. We will make use of novel tools (GILZ transgenic and knockout mice and MaxCyte lentiviral transfection of primary human and murine T cells) to manipulate GILZ expression in ERβ-deficient versus – sufficient Tregs in order to understand how intact ERβ signaling influences the expression (Subaim 1a) and function (Subaim 1b) of Treg-specific GILZ. Leveraging a large cohort of CD patients available through the Cleveland Digestive Disease Research Core Center (DDRCC), we will apply single-cell RNA sequencing to CD patient mucosal Tregs (discarded surgical samples) in order to identify unique transcriptional signatures (Subaim 1c) associated with ERβlow/GILZhigh Tregs. These assays will provide critical data to inform future proposals focused on the intersection of E2 signaling and GILZ activity in Tregs. Successful completion of this proposal will provide mechanistic insight into the regulation of Treg GILZ expression and function via estrogen signaling. These studies will determine the link between diminished ERβ signaling, characteristic of many auto- inflammatory diseases including IBD, and glucocorticoid signaling, with an ultimate goal of developing improved personalized medicine approaches for CD patients.

项目总结/摘要糖皮质激素诱导的亮氨酸拉链（GILZ）是一种广泛表达的转录因子，其表达与糖皮质激素诱导的亮氨酸拉链的表达相关。通过糖皮质激素受体（GR）的内源性或合成糖皮质激素（GC）信号传导诱导。 GILZ被认为介导了GCs在T淋巴细胞中的许多抗炎作用，如诱导TGFβ 信号传导和NFκB、MAPK和AP-1通路的抑制。CD 4 + T细胞中GILZ的条件性缺失导致增强实验性二硝基苯磺酸（DNBS）诱导的结肠炎，一种Th 1介导的实验性炎症性肠病（IBD）。类固醇激素，包括皮质类固醇和性类固醇激素如17β-雌二醇（雌激素，E2）在受体结合中表现出相当大的混杂性。E2 已显示拮抗人子宫上皮细胞中的GILZ，但关于GILZ的作用知之甚少。它可能有助于T细胞中GILZ表达和/或功能的机制。我们的初步数据揭示了在分离自人的调节性T细胞（TCLs）中在mRNA和蛋白质水平上对GILZ的稳健诱导，克罗恩病（CD）患者和SAMP/YitFC（“SAMP”）小鼠，克罗恩病样疾病的自发模型。在这些乳腺癌中，GILZ上调（“GILZhigh”）与雌激素受体β的相对减少有关。 (ERβ）的表达，表明TcB中GILZ的表达可能响应于雌激素信号传导。此外，与GILZlow对应物相比，GILZhigh Tactide表现出降低的抑制功能，这表明，在Tendon中最佳的GILZ介导的免疫抑制可能需要完整的ERβ信号传导。几种慢性炎性和自身免疫性疾病表现出ERβ表达和/或活性的降低，这导致了一种有趣的可能性，即ERβ表达的减少通过破坏细胞的增殖而导致炎症。正常保护性Treg GILZ介导的机制。我们的假设是Treg的保护功能- 特异性GILZ需要完整ERβ信号传导，因此不能介导足够的免疫保护， ERβ耗尽的环境，如IBD肠道。该项目的目标是确定降低ERβ特异性信号传导影响IBD中GILZ表达和功能的机制- 相关的Tibet。我们将利用新的工具（GILZ转基因和基因敲除小鼠和MaxCyte慢病毒转染原代人和鼠T细胞），以操纵ERβ缺陷型与ERβ缺陷型小鼠中GILZ的表达。足够的TdR，以了解完整的ERβ信号传导如何影响表达（Subaim 1a）， Treg特异性GILZ的功能（Subaim 1b）。利用可通过以下途径获得的大量CD患者队列克利夫兰消化疾病研究核心中心（DDRCC），我们将应用单细胞RNA测序CD 患者粘膜THBE（丢弃的手术样品），以鉴定独特的转录签名（Subaim 1c）与ERβ低/GILZ高TdR相关。这些分析将提供关键数据，以告知未来提案集中在Tibetan中E2信号传导和GILZ活性的交叉点上。成功完成本该提案将提供通过雌激素调节Treg GILZ表达和功能的机制见解发信号。这些研究将确定减少的ERβ信号传导之间的联系，这是许多自体免疫系统的特征。炎症性疾病，包括IBD和糖皮质激素信号传导，最终目标是开发改善的为CD患者提供个性化的治疗方法。