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

• 批准号: 10704686
• 负责人: Suthat Liangpunsakul
• 金额: $ 35.66万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 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/10704686
• 关键词: 5' Untranslated Regions; Address; Alcohol consumption; Alcoholic Liver Cirrhosis; Alcoholic Liver Diseases; Alcohols; Animal Model; CXCL1 gene; Cell model; Cells; 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; Stress; 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; stressor; transcriptome sequencing; upstream kinase

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.

项目摘要 酒精相关的肝病（ALD）是一种复杂的疾病。它的发病机理是一个多步骤的过程 通过各种组织病理学变化进行。 FK506结合蛋白-51（FKBP51，由编码 FKBP5基因，也称为FKBP5）属于FKBP免疫蛋白家族。 FKBP5很重要 蛋白质参与了许多关键细胞信号级联反应的调节。我们观察到了两个倍 与健康相比 控件。在乙醇喂养中还观察到了转录本和蛋白质水平的FKBP5表达的增加 老鼠。有趣的是，FKBP5损失受到抗酒精诱导的肝损伤的影响。我们的总体目标是 进一步了解FKBP5在介导酒精诱导的肝损伤中的机械作用。实现 这个目标，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 酒精性肝硬化患者在其启动子的FKBP5甲基化水平上有明显的下调 地区。在第一个特定目标中，我们将仔细剖析乙醇如何诱导FKBP5的上游途径 通过测试以下假设的表达，即位于其CPG岛的FKBP5甲基化水平降低 5'UTR启动子区域通过乙醇导致其转录本和蛋白质表达的增加。接下来，我们会的 确定酒精-FKBP5轴的下游分子机制在酒精诱导的肝损伤中。 我们的数据表明，在用乙醇喂食的野生型小鼠中，河马途径被关闭，如 YAP的去磷酸化（可能通过P-MST1/2的降低，其上游激酶）导致YAP 核易位；在用乙醇喂食的FKBP5 - / - 小鼠中废除的观察结果。我们会谨慎 确定FKBP5和YAP磷酸化的机械作用，作为探索探索的重要步骤 酒精-FKBP5轴的下游分子机制在酒精诱导的肝损伤中。最后， 公开可用的单细胞RNASEQ数据表明肝细胞中FKBP5的差异表达 和其他非羊皮副细胞，尤其是库普弗细胞。解决细胞类型特异性的作用 FKBP5在ALD的发病机理中，我们成功地生成了一种新型的FKBP5 FL/FL小鼠模型 CHRISPR/CAS9策略。我们的初步数据说明乙醇可以诱导FKBP5的表达 与乙醇处理的库普弗细胞具有显着诱导的肝细胞和kupffer细胞相比 肝细胞。我们将检验以下假设，即FKBP5对我们的全球影响ALD的受保护作用 敲除模型是使用Kupffer细胞使用Kupffer特异性FKBP5 - / - 鼠标模型驱动的。拍摄 我们一起开发了动物和细胞模型，以机械检查上游和下游 FKBP5在ALD发病机理中的作用的途径。该提议具有重要意义，可能导致 通过靶向FKBP5鉴定ALD患者的潜在治疗干预措施。