Role of Ceramide in Hepatic Stellate Cell Activation and Liver Fibrosis

神经酰胺在肝星状细胞活化和肝纤维化中的作用

• 批准号: 10222658
• 负责人: Jennifer Y. Chen
• 金额: $ 18.36万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2023-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10222658
• 关键词: Address; Advisory Committees; Amino Acids; Award; Basic Science; Carbon Tetrachloride; Cell Differentiation process; Cells; Ceramides; Chemicals; Choline; Cicatrix; Complement; Cytoplasm; Data; Development; Disease Progression; Ensure; Environment; Enzyme Inhibition; Enzymes; Extracellular Matrix; Farber' s lipogranulomatosis; Fibroblasts; Fibrosis; Funding; Gastroenterologist; Gene Expression; General Hospitals; Genes; Genetic; Genetic Transcription; Goals; Hepatic Fibrogenesis; Hepatic Stellate Cell; High Fat Diet; Human; Immunoblotting; Knowledge; Life; Lipids; Liver Failure; Liver Fibrosis; Liver diseases; Massachusetts; Mechanics; Mediating; Mentorship; Metabolism; Methodology; Mission; Modeling; Molecular; Molecular Biology; Mus; Myofibroblast; Nortriptyline; Pathway interactions; Patients; Pharmacology; Phenotype; Phosphorylation; Physicians; Production; Proteins; Public Health; Public Health Schools; Publications; RNA Interference; Research; Research Personnel; Rest; Role; Scientist; Signal Pathway; Signal Transduction; Sphingolipids; Supervision; Techniques; Therapeutic; Time; Training; Transgenic Mice; Tricyclic Antidepressive Agents; United States National Institutes of Health; Work; career; career development; cell type; chronic liver disease; chronic liver injury; collaborative environment; college; experience; fibrogenesis; galactosylgalactosylglucosylceramidase; genome-wide; genome-wide analysis; improved; in vivo; in vivo Model; liver injury; medical schools; mouse model; multidisciplinary; mutant; non-alcoholic fatty liver disease; novel; novel therapeutics; prevent; programs; response; small molecule; small molecule inhibitor; success; supportive environment; targeted treatment

PROJECT SUMMARY/ABSTRACT This is an application for a K08 award for Dr. Jennifer Y. Chen, a gastroenterologist and hepatologist at the Massachusetts General Hospital (MGH). Dr. Chen’s long-term career goal is to become an independently funded physician scientist, devoting more than 75% of her time to establish and maintain a basic science research program in hepatic fibrosis. This K08 award will provide Dr. Chen with the support necessary to achieve her short-term goals: 1) develop additional training in molecular biology, including investigating protein interactions and induction of stable gene expression; 2) gain expertise in genome-wide transcriptional analysis; 3) become proficient in transgenic mouse work and mouse models of hepatic fibrosis; and 4) produce the data and publication record necessary for a successful R01 application. In prior work, the candidate has developed a small molecule screen to identify compounds that inactivate hepatic stellate cells (HSCs), the primary cell type responsible for hepatic fibrosis. Through analysis of a hit, she identified that the sphingolipid ceramide can profoundly inhibit the activated effector phenotype of HSCs. Her work has identified a potential antifibrotic role for inhibition of acid ceramidase (aCDase), the enzyme responsible for ceramide metabolism. In this proposal, the candidate seeks to elucidate the mechanism by which ceramide accumulation inactivates HSCs and define the role of aCDase in fibrosis progression. The specific goals of the study are to: 1) determine how ceramide mediates HSC inactivation; and 2) define the impact of aCDase depletion and inhibition on fibrosis development in vivo. Under the first aim, the applicant will utilize a combination of techniques including immunoblotting, small molecule inhibition and activation, RNA interference, and expression of constitutively activated mutant proteins to understand how ceramide inactivates HSCs. Genome-wide expression analysis will be performed to elucidate transcriptional targets of ceramide. Under the second aim, the applicant will generate mice with a conditional fibroblast-specific deletion of aCDase, and will determine the extent to which they experience reduced fibrosis in two independent models of hepatic fibrosis. She will also determine the antifibrotic effects of a small molecule inhibitor of aCDase in vivo. As an integral part of this proposal, the candidate’s career development will be complemented by participation in advanced coursework and research seminars to develop expertise in molecular biology, genome-wide transcriptional analysis, and in vivo models of hepatic fibrosis. A formal mentorship committee and advisory team will provide supervision, guidance, and assistance for the candidate to achieve her goals. The research environment, which includes the MGH GI Unit, Harvard Fibrosis Network, Harvard College, Harvard Medical School, and the Harvard School of Public Health, will provide a rich, collaborative, and supportive atmosphere to ensure the candidate’s success. Through this award, the candidate will become an independent basic science investigator by contributing to the understanding of the role of the sphingolipid ceramide and its inactivation of HSCs ex vivo and in vivo.

项目摘要/摘要 这是詹妮弗·乔（Jennifer Y. 马萨诸塞州综合医院（MGH）。陈博士的长期职业目标是成为独立的 资助的物理科学家，将超过75％的时间用于建立和维护基础科学 肝纤维化研究计划。该K08奖将为Chen博士提供必要的支持 实现她的短期目标：1）在分子生物学方面开发额外的培训，包括研究蛋白质 稳定基因表达的相互作用和诱导； 2）在全基因组转录分析方面获得专业知识； 3）精通转基因小鼠的工作和肝纤维化的小鼠模型； 4）产生数据 和成功的R01应用程序所需的出版记录。在先前的工作中，候选人已经发展 一个小的分子筛选，以识别灭活肝星状细胞（HSC）的化合物（主要细胞） 负责肝纤维化的类型。通过对命中的分析，她确定鞘脂神经酰胺可以 深刻抑制HSC的活化效应子表型。她的工作已经确定了潜在的抗纤维化角色 为了抑制酸神经酰胺酶（ACDASE），负责神经酰胺代谢的酶。在此提案中， 候选人旨在阐明神经酰胺积累灭活HSC并定义的机制 ACDase在纤维化进展中的作用。研究的具体目标是：1）确定神经酰胺 介导HSC失活； 2）定义ACDase部署和抑制对纤维化的影响 体内发展。在第一个目标下，申请人将利用包括 免疫印迹，小分子抑制和激活，RNA干扰和组成性的表达 活化的突变蛋白了解神经酰胺如何使HSC失活。全基因组表达分析 将执行以阐明神经酰胺的转录靶标。在第二个目标下，适用的将 用条件成纤维细胞特异性删除ACDase产生小鼠，并将确定多大程度 在两个独立的肝纤维化模型中，它们的纤维化降低。她还将确定 体内ACDase的小分子抑制剂的抗纤维化作用。作为本提案不可或缺的一部分， 候选人的职业发展将通过参与高级课程和研究完成 培养分子生物学，全基因组转录分析和体内模型方面的专业知识 肝纤维化。正式的攻表委员会和咨询小组将提供监督，指导和 候选人实现她的目标的帮助。研究环境，包括MGH GI单元， 哈佛大学纤维化网络，哈佛大学，哈佛医学院和哈佛公共卫生学院， 将提供丰富，协作和支持的氛围，以确保候选人的成功。通过这个 奖项，候选人将通过为 了解鞘脂神经酰胺的作用及其对HSC的灭活，体内和体内的作用。