Cholesterol toxicity in alcohol-associated hepatitis

酒精相关性肝炎中的胆固醇毒性

• 批准号: 10637154
• 负责人: Zhenghui Gordon Jiang
• 金额: $ 70.15万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-15 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10637154
• 关键词: Acute; Acyltransferase; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcohols; Atlases; Bile Acid Biosynthesis Pathway; Bile Acids; Binding; Blood; Cells; Cholestasis; Cholesterol; Cholesterol 7-alpha-Monooxygenase; Cholesterol Homeostasis; Chronic; Development; Disease; Disease model; Enzymes; Esterification; Exhibits; Experimental Models; Functional disorder; HNF4A gene; Hepatitis; Hepatocyte; Hepatotoxicity; Human; Impairment; In Vitro; Individual; Knowledge; Lecithin; Link; Lipoproteins; Liver; Liver Failure; Liver diseases; Maps; Messenger RNA; Metabolic Pathway; Modeling; Mus; Nuclear RNA; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Production; Protein Isoforms; Proteins; RNA; RNA Interference; Regulatory Pathway; Resolution; Route; Sampling; Scientist; Technology; Testing; Therapeutic; Toxic effect; Transgenic Model; Transportation; Treatment Efficacy; Up-Regulation; Very low density lipoprotein; efficacy testing; feeding; first-in-human; hepatocyte nuclear factor; in vivo; mortality; mouse model; multidisciplinary; multimodality; multiplexed imaging; novel; novel therapeutic intervention; pre-clinical; predict clinical outcome; reverse cholesterol transport; screening; success; targeted treatment; therapeutic target; tool; transcription factor; transcriptome sequencing; transcriptomics; treatment strategy; uptake

SUMMARY Alcohol-associated hepatitis (AH) is a form of subacute liver failure with high mortality, limited treatment options, and elusive pathogenesis. Emerging evidence suggests that AH is associated with a profound impairment in hepatocellular cholesterol homeostasis. Hepatocytes are the central hub for cholesterol metabolism. Meanwhile, AH hepatocytes exhibit profoundly altered FC content and pathways responsible for cholesterol esterification, transport, and conversion to bile acids. RNA sequencing studies indicate that the dysregulation of HNF4α, a master hepatocyte transcription factor, is responsible for impaired cholesterol homeostasis. Upregulation of HNF4α using small activating RNA (saRNA) could mitigate cholesterol toxicity in vitro and can be developed as a treatment for AH. Our long-term objective is to define the disease-centric pathways of AH and develop much-needed therapeutics for AH patients. We hypothesize that the HNF4a-dependent impairment of cholesterol homeostasis in hepatocytes directly contributes to hepatocellular dysfunction in AH and can be targeted therapeutically. Our approach is to map regulatory pathways of cholesterol metabolism in human AH at a single cell resolution, functionally interrogate cholesterol metabolic pathways in murine alcohol models, and test a novel therapeutic strategy by targeting HNF4a-dependent cholesterol toxicity. Aim 1 will map cholesterol metabolism in AH at single cell resolution using a multimodal approach. A three- pronged strategy will map AH-associated changes in cholesterol homeostasis using single nuclear RNA sequencing, ultra-high multiplexed imaging, and targeted spatial transcriptomics. The unbiased reference maps will guide the interrogation of cholesterol metabolic pathways in cell and murine AH models. Aim 2 will define the impact of cholesterol dysregulation on alcohol-induced liver injury in murine models. We will model cholesterol toxicity in 3 conditions relevant to AH using the chronic-plus-binge alcohol feeding: (1) acute cholesterol overload, (2) impaired bile acid synthesis, and (3) impaired cholesterol secretion in cholestasis. This effort will generate optimized murine models that recapitulate cholesterol toxicity in AH. Aim 3 will evaluate targeted manipulation of HNF4α-dependent pathways of cholesterol toxicity as a novel therapeutic strategy in AH. We use saRNA to modulate HNF4α isoform expression and cholesterol metabolism. GalNAc-conjugated HNF4α-saRNA will be tested as a hepatocyte-targeted therapeutics to mitigate cholesterol toxicity in models of AH using differentiated human hepatocytes and murine models. This project is built upon a strong team of multidisciplinary scientists combining unique expertise in lipidology, preclinical liver models, and state-of-the-art single cell and spatial technologies. The success of this project will further our understanding of AH pathogenesis and could lead to the development of a novel AH treatment ready for preclinical and first-in-human trials.

总结 酒精相关性肝炎（AH）是一种亚急性肝功能衰竭，死亡率高，治疗有限 选择和难以捉摸的发病机制。新出现的证据表明，AH与一种深刻的 肝细胞胆固醇稳态受损。肝细胞是胆固醇的中心枢纽 新陈代谢.与此同时，AH肝细胞表现出FC含量的深刻改变和负责的途径 胆固醇酯化、转运和转化为胆汁酸。RNA测序研究表明， HNF 4 α是一种主要的肝细胞转录因子，其调节异常导致胆固醇受损 体内平衡使用小激活RNA（saRNA）上调HNF 4 α可以减轻胆固醇毒性， 体外，并可开发为AH的治疗。 我们的长期目标是确定AH的疾病中心途径，并开发急需的 治疗AH患者。我们假设HNF 4a依赖性胆固醇损伤 肝细胞内的稳态直接导致AH中的肝细胞功能障碍， 治疗上我们的方法是绘制人类AH中胆固醇代谢的调节途径， 单细胞分辨率，在小鼠酒精模型中功能性地询问胆固醇代谢途径，以及 通过靶向HNF 4a依赖性胆固醇毒性来测试新的治疗策略。 目标1将使用多模式方法以单细胞分辨率绘制AH中的胆固醇代谢。一个三- 一项有针对性的策略将使用单核RNA绘制AH相关的胆固醇稳态变化 测序、超高多重成像和靶向空间转录组学。无偏参考 图将指导细胞和小鼠AH模型中胆固醇代谢途径的询问。 目的2将明确胆固醇失调对酒精诱导的小鼠肝损伤模型的影响。 我们将使用慢性加酗酒喂养在3种与AH相关的条件下模拟胆固醇毒性： (1)急性胆固醇超载，（2）胆汁酸合成受损，和（3）胆固醇分泌受损 胆汁淤积这项工作将产生最佳的小鼠模型，重现AH中的胆固醇毒性。 目的3将评价靶向操纵HNF 4 α依赖的胆固醇毒性途径作为一种新的 治疗策略。我们使用saRNA调节HNF 4 α亚型表达和胆固醇水平， 新陈代谢. GalNAc偶联的HNF 4 α-saRNA将作为肝细胞靶向治疗剂进行测试， 在使用分化的人肝细胞和鼠模型的AH模型中减轻胆固醇毒性。 该项目是建立在一个强大的多学科科学家团队结合独特的专业知识， 脂质学、临床前肝脏模型以及最先进的单细胞和空间技术。的成功 该项目将进一步加深我们对AH发病机制的理解，并可能导致开发一种新的AH 为临床前和首次人体试验做好准备。