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; 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.

项目摘要 /摘要 肝硬化肝病是美国死亡率的第11个主要原因，没有治疗 肝移植以外的阶段疾病。因此，该提案的总体目的是阐明新颖 驱动纤维化信号释放的机制导致肝纤维化进展并指导 发展潜在的治疗策略。肝纤维化的特征是肝星状的激活 细胞（HSC）。我们在原代人和小鼠HSC以及体内的初步数据证明了1。 血小板衍生的生长因子B（PDGF）通过增加糖酵解来诱导代谢重编程； 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（H3K9AC）在 囊泡运输相关的RAS相关蛋白Rab（RAB）基因的启动子区域； 3。糖酵解促进EV 用纤维蛋白释放和富集；和4。通过HSC选择性己糖酶抑制体内糖酵解抑制 2（HK2）缺失消除了肝纤维化。我们已经利用了我们的小说发现来产生中央 当前PDGF介导的HSC中糖酵解的当前建议诱导纤维源EV 通过H3K9AC依赖性的转录上调释放，以扩大肝纤维化。我们将 采用复杂的细胞和动物模型，包括DCAS-KRAB模型的体外和体内利用 体内HSC特异性HK2缺失模型以及乙酰-COA缺陷HSC，以研究以下 集成但独立的目标。在AIM 1中，我们将检验以下假设：PDGF通过 赖氨酸缺陷型激酶1（WNK1）磷酸化以介导葡萄糖转运蛋白1（GLUT1）易位 质膜。我们将揭示导致HSC中糖酵解的激酶信号传导：研究PDGF 通过WNK1的磷酸化来增加糖酵解，WNK1是一种新型的PDGF下游赖氨酸缺陷激酶。和 b。研究WNK1磷酸化如何促进GLUT1易位到质膜以增加 糖酵解，代表HSC中的一种新机制。在AIM 2中，我们将测试糖酵解的假设 通过上调通过乙酰辅酶A（乙酰-COA）介导的RAB的转录来释放EV H3K9AC。我们将剖析糖酵解如何驱动囊泡运输基因计划的表观遗传调节以控制 EV发布者：研究PDGF如何促进代谢物乙酰辅酶A的积累以增加 H3K9AC;和b。检查H3K9AC如何促进RAB转录以诱导EV释放。在AIM 3中，我们将 检验HSC特异性糖酵解和随后的表观遗传调节的假设 放大体内肝纤维化。我们将通过以下方式研究体内肝纤维化扩增的机制。 研究糖酵解介导的EV如何通过靶向HSC来扩增肝纤维化；和b。调查如何 RAB的表观遗传调节的破坏可以改善肝纤维化。这本小说和创新的探究线 将定义HSC特异性糖酵解依赖性肝纤维化扩增模型，并将轨迹朝向 治疗人类肝纤维化和肝硬化的新的和重大的进步。