FKBP5 in the pathogenesis of alcohol-associated liver disease

FKBP5 在酒精相关性肝病发病机制中的作用

基本信息

批准号：
10501012
负责人：
Suthat Liangpunsakul
金额：
$ 35.03万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10501012
关键词：
5&apos Untranslated Regions Address Alcohol consumption Alcoholic Liver Cirrhosis Alcoholic Liver Diseases Alcohols Animal Model CXCL1 gene Cell model Chromosome 6 Chromosome Arm Complex CpG Islands Data Diet Disease Down-Regulation Ethanol FK506 binding protein 5 Family Gene Expression Genes Goals Health Hepatocyte Immunophilins Knock-out Knockout Mice Kupffer Cells Lead Link Liver Liver diseases Mediating Mental Depression Mental disorders Metabolic Methylation Modeling Molecular Molecular Chaperones Mus Nuclear Translocation Pathogenesis Pathway interactions Patients Phosphorylation Post-Traumatic Stress Disorders Process Promoter Regions Protein Dephosphorylation Proteins Public Health Regulation Risk Factors Role Sampling Secondary to Series Signal Transduction Tacrolimus Binding Proteins Testing Therapeutic Intervention Transcript United States Up-Regulation Wild Type Mouse alcohol related problem alcohol risk biological adaptation to stress cell type chronic liver disease differential expression drinking epigenetic regulation insight liver transplantation mRNA Expression mouse model novel novel therapeutic intervention protective effect protein expression single-cell RNA sequencing stress related disorder stressor transcriptome sequencing upstream kinase

项目摘要

Project Summary Alcohol-associated liver disease (ALD) is a complex disorder; its pathogenesis is a multi-step process that progresses through a spectrum of histopathological changes. FK506-binding protein-51 (FKBP51, encoded by the FKBP5 gene, also called FKBP5) belongs to the FKBP family of immunophilins. FKBP5 is an important protein involved in the regulation of many key cellular signaling cascades. We observed a two-fold upregulation of FKBP5 mRNA expression in the liver of patients with alcoholic cirrhosis compared to healthy controls. The increase in Fkbp5 expression at the transcript and protein level was also observed in ethanol-fed mice. Interestingly, loss of Fkbp5 protected against alcohol-induced liver injury. Our overarching goal is to further understanding the mechanistic action of FKBP5 in mediating alcohol-induced liver injury. To achieve this goal, we will first determine how alcohol induces FKBP5 expression. Our preliminary data suggested that patients with alcoholic cirrhosis had significant downregulation in FKBP5 methylation levels at its promoter region. In the first specific aim, we will carefully dissect the upstream pathway of how ethanol induces FKBP5 expression by testing the hypothesis that reduced methylation levels of Fkbp5 at the CpG island located at its 5’ UTR promoter region by ethanol lead to an increase in its transcript and protein expression. Next, we will determine the downstream molecular mechanism of the alcohol-FKBP5 axis in alcohol-induced liver injury. Our data suggested that in wild-type mice fed with ethanol, the Hippo pathway was turned off as indicated by dephosphorylation of YAP (likely through the reduction in p-MST1/2, its upstream kinase) leading to YAP nuclear translocation; the observation which was abrogated in Fkbp5-/- mice fed with ethanol. We will carefully determine the mechanistic role of Fkbp5 and Yap phosphorylation, as an essential step in exploring the downstream molecular mechanism of the alcohol-FKBP5 axis in alcohol-induced liver injury. Lastly, the publicly available single-cell RNASeq data suggested the differential expression of FKBP5 in the hepatocytes and other non-hepatic parenchymal cells, notably Kupffer cells. To address the role of cell-type-specific FKBP5 in the pathogenesis of ALD, we successfully generated a novel Fkbp5 fl/fl mouse model using the CHRISPR/Cas9 strategy. Our preliminary data illustrated that ethanol can induce the expression of Fkbp5 in both hepatocytes and Kupffer cells with a significant induction in Kupffer cells treated with ethanol compared to that of hepatocytes. We will test the hypothesis that the protective effect of Fkbp5 against ALD in our global knockout model is driven primarily by Kupffer cells using Kupffer cell-specific Fkbp5-/- mouse model. Taken together, we have developed animal and cellular models to mechanistically examine both up- and downstream pathways on the role of FKBP5 in ALD pathogenesis. This proposal is of significance and it may lead to the identification of potential therapeutic interventions in patients with ALD by targeting FKBP5.

项目概要酒精相关性肝病 (ALD) 是一种复杂的疾病，其发病机制是一个多步骤的过程， FK506 结合蛋白 51（FKBP51，由 FK506 编码）发生一系列组织病理学变化。 FKBP5基因（也称为FKBP5）属于亲免素FKBP家族，FKBP5是一个重要的基因。我们观察到参与许多关键细胞信号级联调节的蛋白质。与健康人相比，酒精性肝硬化患者肝脏中 FKBP5 mRNA 表达上调对照组中 Fkbp5 在转录物和蛋白质水平上的表达也有所增加。提示，Fkbp5 的缺失可以防止酒精引起的肝损伤。进一步了解FKBP5介导酒精性肝损伤的机制。为了实现这一目标，我们将首先确定酒精如何诱导 FKBP5 表达。我们的初步数据表明。酒精性肝硬化患者 FKBP5 启动子甲基化水平显着下调在第一个具体目标中，我们将仔细剖析乙醇如何诱导 FKBP5 的上游途径。通过测试降低位于其 CpG 岛的 Fkbp5 甲基化水平的假设来表达表达乙醇的 5'UTR 启动子区域导致其转录物和蛋白质表达增加。确定酒精-FKBP5轴在酒精性肝损伤中的下游分子机制。我们的数据表明，在喂食乙醇的野生型小鼠中，Hippo 通路被关闭，如下所示： YAP 去磷酸化（可能通过其上游激酶 p-MST1/2 的减少）导致 YAP 核易位；在用乙醇喂养的Fkbp5-/-小鼠中观察到的结果被消除。确定 Fkbp5 和 Yap 磷酸化的机制作用，作为探索酒精-FKBP5轴在酒精性肝损伤中的下游分子机制。公开的单细胞 RNASeq 数据表明 FKBP5 在肝细胞中的差异表达和其他非肝实质细胞，特别是库普弗细胞，以解决细胞类型特异性的作用。 FKBP5 在 ALD 发病机制中的作用，我们使用我们的初步数据表明乙醇可以诱导 Fkbp5 的表达。与用乙醇处理的库普弗细胞相比，肝细胞和库普弗细胞均具有显着诱导作用我们将测试 Fkbp5 对全球肝细胞的保护作用这一假设。敲除模型主要由 Kupffer 细胞驱动，采用 Kupffer 细胞特异性 Fkbp5-/- 小鼠模型。我们共同开发了动物和细胞模型来机械地检查上游和下游 FKBP5 在 ALD 发病机制中的作用途径这一建议具有重要意义，可能会导致通过靶向 FKBP5 鉴定 ALD 患者的潜在治疗干预措施。