Progression from steatosis to steatohepatitis in alcoholic liver disease is enhanced by the risk allele of PNPLA3 via remodeling of lipid droplets and disruption of bioactive lipid composition

PNPLA3 的风险等位基因通过脂滴重塑和生物活性脂质成分破坏，增强了酒精性肝病从脂肪变性到脂肪性肝炎的进展

• 批准号: 9979504
• 负责人: Paulina M Ordonez Naranjo
• 金额: $ 22.64万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-10 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9979504
• 关键词: Ablation; Adipose tissue; Affect; Alcohol Phenotype; Alcohol consumption; Alcoholic Liver Diseases; Alcohols; Amino Acids; Arachidonate 15-Lipoxygenase; Arachidonate 5-Lipoxygenase; Arachidonic Acids; Attenuated; Biology; CD59 Antigen; Cause of Death; Cell Line; Cell physiology; Cellular Stress; Cessation of life; Characteristics; Chronic; Country; Cyclooxygenase Inhibitors; Development; Disease; Disease Progression; Disease model; Docosahexaenoic Acids; Economics; Eicosanoids; Eicosapentaenoic Acid; Engineering; Enzymes; Fatty Acids; Fatty Liver; Fatty acid glycerol esters; Fibrosis; Flow Cytometry; Genes; Genetic; Genetic Polymorphism; Genetic Predisposition to Disease; Genome; Goals; Hepatitis; Hepatocyte; Hepatotoxicity; Human; IL6 gene; Imaging Techniques; In Situ; Inflammation; Inflammation Mediators; Inflammatory; Isoleucine; Knock-out; Knowledge; LOX gene; Lead; Leukotrienes; Lipids; Lipolysis; Lipoxins; Liver; Liver Fibrosis; Liver diseases; Mass Spectrum Analysis; Measurement; Measures; Mediating; Medication Management; Metabolic; Methionine; Microscopy; Morphology; Mus; Nonesterified Fatty Acids; Organelles; Palmitic Acids; Patients; Pharmacology; Pharmacotherapy; Phospholipids; Physiology; Population; Predisposition; Prostaglandins; Proteins; Proteomics; Protocols documentation; Public Health; Publishing; Research Project Grants; Steatohepatitis; Stress; Subcellular Fractions; TNF gene; Technology; Testing; Thromboxanes; Triglycerides; United States; United States National Institutes of Health; Variant; Western Blotting; alcohol exposure; base; cellular imaging; chronic alcohol ingestion; chronic liver disease; cytokine; design; experimental study; gene induction; genetic risk factor; genetic variant; human model; human stem cells; induced pluripotent stem cell; liver injury; new therapeutic target; non-alcoholic fatty liver disease; overexpression; programs; response; risk variant; stem cell differentiation; therapeutic target; transcription activator-like effector nucleases

Project Summary/Abstract: Alcoholic liver disease (ALD) is one of the main causes of chronic liver disease worldwide and accounts for up to 48% of deaths associated with end stage liver disease in the United States 1. Compared to other types of liver disease, there has been little progress in the pharmacological management of ALD. Factors contributing to a lack of specific therapeutic targets include current suboptimal disease models and the gap in our knowledge of mechanisms of susceptibility and tolerance to chronic alcohol exposure. We propose to bridge this gap by using human stem cell and gene editing technology to study the most important genetic susceptibility factor involved in ALD progression; the risk variant polymorphism of PNPLA3 2-4. The variant allele of PNPLA3 (148 isoleucine to methionine protein variant; I148M) induces abnormal lipolysis and contributes to enhanced hepatic steatosis as a result of chronic alcohol consumption, as well as more aggressive progression to inflammation and fibrosis that are characteristic of the advanced stages of ALD2,4. We have already generated the human induced pluripotent stem cell lines that will be used for this proposal, and we have optimized a protocol to differentiate these stem cells to hepatocytes. We have performed basic characterization of PNPLA3 variant hepatocytes and confirmed that they accumulate triglyceride-rich lipid droplets and express inflammatory cytokines that are thought to be involved in progression from steatosis to hepatitis. To understand the mechanism by which the variant PNPLA3 confers increased susceptibility to alcohol-related liver injury, we will perform detailed profiling of lipid droplets of hepatocytes derived from human stem cell lines engineered to express the risk variant of PNPLA3. We will determine the lipid composition, protein composition and profile of specific inflammatory mediators that determine progression from steatosis to inflammation in ALD patients. We will use ethanol and palmitic acid to mimic the environmental stress induced by alcohol consumption and lipotoxicity. Our main goal is to identify novel therapeutic targets directly in live human hepatocytes that can be exploited for the treatment of ALD.

项目摘要/摘要： 酒精性肝病（ALD）是全世界慢性肝病的主要原因之一，占全球慢性肝病的主要原因之一。 在美国，48% 的死亡与终末期肝病相关 1. 与其他类型的肝脏相比 疾病，ALD 的药物治疗进展甚微。促成因素 缺乏具体的治疗靶点包括当前的次优疾病模型以及我们对疾病的了解方面的差距 对长期酒精暴露的易感性和耐受性机制。我们建议通过使用来弥补这一差距 人类干细胞和基因编辑技术研究涉及的最重要的遗传易感因素 在 ALD 进展中； PNPLA3 2-4 的风险变异多态性。 PNPLA3 的变异等位基因（148 异亮氨酸 蛋氨酸蛋白变体； I148M）诱导异常脂肪分解并导致肝脏脂肪变性增强 由于长期饮酒以及炎症和纤维化更加严重的进展 这是 ALD2,4 高级阶段的特征。我们已经生成了人类诱导的 将用于该提案的多能干细胞系，我们已经优化了一个方案来区分 这些干细胞转化为肝细胞。我们已经对 PNPLA3 变异肝细胞进行了基本表征，并 证实它们积累富含甘油三酯的脂滴并表达炎症细胞因子 据认为参与从脂肪变性到肝炎的进展。了解该机制 变异 PNPLA3 增加对酒精相关肝损伤的易感性，我们将进行详细分析 源自人类干细胞系的肝细胞脂滴，被设计用于表达风险变异 PNPLA3。我们将确定特定炎症的脂质成分、蛋白质成分和概况 决定 ALD 患者从脂肪变性进展为炎症的介质。我们将使用乙醇和 棕榈酸模拟由饮酒和脂毒性引起的环境压力。我们的主要目标 是直接在活人肝细胞中识别可用于治疗的新治疗靶点 酒精性肝病。