Pathobiology of liver fibrosis

肝纤维化的病理学

• 批准号: 10711085
• 负责人: Enis Kostallari
• 金额: $ 35.51万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711085
• 关键词: AGFG1 gene; Acetyl Coenzyme A; Acetylation; Animal Model; Bar Codes; Biological Assay; Cell membrane; Cell model; Cellular Metabolic Process; Cirrhosis; Data; Development; Disease; Epigenetic Process; Fibrosis; Gene Expression; Genes; Genetic Transcription; Genus Hippocampus; Glycolysis; Glycolysis Inhibition; Hepatic Stellate Cell; Hexokinase 2; Histones; Human; In Vitro; Knowledge; Liver; Liver Cirrhosis; Liver Fibrosis; Liver diseases; Lysine; Mediating; Metabolic; Modeling; Mus; Outcome; Pathologic; Phosphorylation; Phosphotransferases; Platelet-Derived Growth Factor B; Play; Promoter Regions; Proteins; RAS Superfamily Proteins; Regulation; Research; Research Proposals; Role; SLC2A1 gene; Signal Pathway; Signal Transduction; Spatial Distribution; Technology; Testing; Therapeutic; Transcriptional Activation; Up-Regulation; Vesicle; Work; effective therapy; epigenetic regulation; extracellular vesicles; glucose metabolism; in vivo; innovation; liver transplantation; mortality; nanoflow cytometry; novel; novel therapeutic intervention; pharmacologic; preclinical study; programs; single-cell RNA sequencing; trafficking; transcriptomics; treatment strategy; vesicular release

PROJECT SUMMARY / ABSTRACT Cirrhotic stage liver disease is the 11th leading cause of mortality in the US, and no treatment exists for late- stage disease other than liver transplantation. Thus, the overall objective of this proposal is to elucidate novel mechanisms that drive the release of fibrogenic signals leading to liver fibrosis progression and to guide the development of potential treatment strategies. Liver fibrosis is characterized by the activation of hepatic stellate cells (HSCs). Our preliminary data in primary human and mouse HSCs as well as in vivo demonstrate that 1. platelet-derived growth factor B (PDGF) induces metabolic reprogramming by increasing glycolysis; 2. PDGF- mediated glycolysis increase the transcriptional activation mark, histone 3 lysine 9 acetylation (H3K9ac) on the promoter region of vesicle trafficking-related Ras-related protein Rab (RAB) genes; 3. glycolysis promotes EV release and enrichment with fibrogenic proteins; and 4. in vivo glycolysis inhibition by HSC-selective hexokinase 2 (HK2) deletion abrogates liver fibrosis. We have utilized our novel findings to generate the CENTRAL HYPOTHESIS of the current proposal that PDGF-mediated glycolysis in HSCs induces fibrogenic EV release through H3K9ac-dependent transcriptional upregulation of RABs to amplify liver fibrosis. We will employ sophisticated cellular and animal models, including in vitro and in vivo utilization of dCas-KRAB model, in vivo HSC-specific HK2 deletion model as well as acetyl-coA-deficient HSCs, to investigate the following integrated, yet independent aims. In Aim 1, we will test the hypothesis that PDGF increases glycolysis through lysine-deficient kinase 1 (WNK1) phosphorylation to mediate glucose transporter 1 (GLUT1) translocation to the plasma membrane. We will uncover the kinase signaling leading to glycolysis in HSCs by: a. studying how PDGF increases glycolysis through phosphorylation of WNK1, a novel PDGF downstream lysine-deficient kinase; and b. investigating how WNK1 phosphorylation promotes GLUT1 translocation to the plasma membrane to increase glycolysis, which represents a new mechanism in HSCs. In Aim 2, we will test the hypothesis that glycolysis leads to EV release by upregulating the transcription of RABs through acetyl coenzyme A (acetyl-CoA)-mediated H3K9ac. We will dissect how glycolysis drives epigenetic regulation of vesicle trafficking gene program to control EV release by: a. studying how PDGF promotes the accumulation of the metabolite acetyl-CoA to increase H3K9ac; and b. examining how H3K9ac promotes RAB transcription to induces EV release. In Aim 3, we will test the hypothesis that HSC-specific glycolysis and subsequent epigenetic regulation of fibrogenic EV release amplifies in vivo liver fibrosis. We will investigate the mechanism of in vivo liver fibrosis amplification by: a. studying how glycolysis-mediated EVs amplify liver fibrosis by targeting HSCs; and b. investigating how the disruption of epigenetic regulation of RABs ameliorates liver fibrosis. This novel and innovative line of inquiry will define an HSC-specific glycolysis-dependent model of liver fibrosis amplification and set a trajectory towards new and significant advances to treat liver fibrosis and cirrhosis in humans.

项目总结/摘要 肝硬化期肝病是美国第11大死亡原因，晚期肝硬化没有治疗方法。 肝移植以外的其他疾病。因此，本提案的总体目标是阐明新颖的 驱动纤维化信号释放导致肝纤维化进展的机制， 开发潜在的治疗策略。肝纤维化的特征是肝星状细胞活化， 细胞（HSC）。我们在原代人和小鼠HSC以及体内的初步数据表明，1。 血小板衍生生长因子B（PDGF）通过增加糖酵解诱导代谢重编程; 2. PDGF- 介导的糖酵解增加了转录激活标记，组蛋白3赖氨酸9乙酰化（H3 K9 ac）， 囊泡运输相关Ras相关蛋白Rab（RAB）基因的启动子区; 3.糖酵解促进EV 释放和富集纤维化蛋白;和4. HSC选择性己糖激酶体内糖酵解抑制 2（HK 2）缺失消除肝纤维化。我们利用我们的新发现来产生中央 目前关于HSC中PDGF介导的糖酵解诱导纤维化EV的假设 通过H3 K9 ac依赖性转录上调释放RAB以放大肝纤维化。我们将 采用复杂的细胞和动物模型，包括体外和体内利用dCas-KRAB模型， 体内HSC特异性HK 2缺失模型以及乙酰辅酶A缺陷型HSC，以研究以下 综合而又独立的目标。在目标1中，我们将检验PDGF通过以下途径增加糖酵解的假设： 赖氨酸缺陷激酶1（WNK 1）磷酸化介导葡萄糖转运蛋白1（GLUT 1）转运到 质膜我们将通过以下方式揭示导致HSC中糖酵解的激酶信号传导：a.研究PDGF如何 通过WNK 1的磷酸化增加糖酵解，WNK 1是一种新的PDGF下游赖氨酸缺陷激酶; B.研究WNK 1磷酸化如何促进GLUT 1转运到质膜， 糖酵解，这代表了HSC中的一种新机制。在目标2中，我们将检验糖酵解 通过乙酰辅酶A（acetyl-CoA）介导的RABs转录上调导致EV释放 H3K9ac。我们将剖析糖酵解如何驱动表观遗传调控囊泡运输基因程序，以控制 EV发行人：A。研究PDGF如何促进代谢产物乙酰辅酶A的积累， H3 K9 ac;和B.研究H3 K9 ac如何促进RAB转录以诱导EV释放。在目标3中，我们 检验HSC特异性糖酵解和随后的纤维化EV释放的表观遗传调节 放大体内肝纤维化。我们将通过以下方法研究体内肝纤维化放大的机制： 研究糖酵解介导的EV如何通过靶向HSC来放大肝纤维化;以及B.调查 破坏RAB的表观遗传调节可改善肝纤维化。这条新颖而创新的研究路线 将定义肝纤维化放大的HSC特异性糖酵解依赖模型，并设定一个 治疗人类肝纤维化和肝硬化的新的和重大的进展。