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.

项目摘要/摘要糖皮质激素诱导的亮氨酸拉链（GILZ）是一种宽阔表达的转录因子，其表达为通过内源性或合成糖皮质激素（GC）信号通过糖皮质激素受体（GR）诱导。吉尔兹被认为会冥想在T淋巴细胞中的许多GCS的抗炎作用，例如诱导TGFβ NFκB，MAPK和AP-1途径的信号传导和抑制。 CD4+ T细胞中GILZ的条件缺失导致增强实验性二硝基苯磺酸（DNB）诱导的结肠炎，一种Th1介导的形式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诸如17β-雌激素（雌激素，E2）之类的术语表现出受体结合的考虑杂交。 E2 已经显示出在人子宫上皮细胞中与吉尔兹拮抗的它可能有助于T细胞中吉尔兹表达和/或功能的机制。我们的初步数据揭示了从从mRNA和蛋白质水平上的gilz诱导的调节性T细胞（Treg）中的蛋白质水平克罗恩病（CD）患者和SAMP/YITFC（“ SAMP”）小鼠，这是克罗恩病的赞助模型。这些Treg中的Gilz上调（“ Gilzhigh”）与雌激素受体β的相对降低有关（ERβ）表达，表明treg中的吉尔兹表达可能对雌激素信号反应。此外，与吉尔兹洛（Gilzlow）相比提示Treg中最佳的GILZ介导的免疫抑制可能需要完整的ERβ信号传导。 ERβ表达和/或活性的几种慢性炎症和自身免疫性疾病的降低，导致ERβ表达降低的引人入胜的可能性会导致炎症通常具有保护的Treg Gilz介导的机制。我们的假设是Treg-的受保护功能特定的吉尔兹需要完整的ERβ信号传导，因此无法介导足够的免疫保护 ERβ缺乏的环境，例如IBD肠。该项目的目的是确定降低ERβ特异性信号传导的机制会影响GILZ在IBD-的表达和功能相关的treg。我们将利用新颖的工具（吉尔兹转基因和淘汰小鼠和maxcyte慢病毒转染原代人和鼠T细胞）以操纵ERβ缺陷与 - 足够的Treg，以了解完整的ERβ信号如何影响表达（Subaim 1a）和 Treg特异性吉尔兹的功能（Subaim 1b）。利用大量通过克利夫兰消化疾病研究核心中心（DDRCC），我们将对CD应用单细胞RNA测序患者粘膜treg（丢弃的手术样本），以识别独特的转录特征（subaim 1c）与erβlow/gilzhigh treg相关联。这些评估将提供关键数据以告知未来提案集中在Treg中E2信号传导和GILZ活性的相交。成功完成提案将通过雌激素对Treg Gilz表达和功能的调节提供机械洞察力信号。这些研究将确定ERβ信号下降之间的联系，这是许多自动的特征包括IBD和糖皮质激素信号在内的炎症性疾病，其最终目标是发展 CD患者的个性化医学方法。