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: 10475911
• 负责人: Jennifer Y. Chen
• 金额: $ 13.77万
• 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/10475911
• 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.

项目概要/摘要 这是 Jennifer Y. Chen 博士的 K08 奖项申请，她是该大学的胃肠病学家和肝病学家 麻省总医院（MGH）陈博士的长期职业目标是成为一名独立的医生。 受资助的医师科学家，投入超过 75% 的时间来建立和维护基础科学 该 K08 奖项将为陈博士提供必要的支持 实现她的短期目标：1）开展分子生物学方面的额外培训，包括研究蛋白质 相互作用和诱导稳定的基因表达；2）获得全基因组转录分析的专业知识； 3) 精通转基因小鼠工作和肝纤维化小鼠模型；4) 生成数据； 候选人在之前的工作中已经制定了成功 R01 申请所需的和发表记录。 一种小分子筛选，用于鉴定使肝星状细胞（HSC）（原代细胞）失活的化合物 通过分析，她发现鞘脂神经酰胺可以导致肝纤维化。 深度抑制 HSC 的激活效应子表型 她的工作已确定了潜在的抗纤维化作用。 用于抑制酸性神经酰胺酶（aCDase），该酶负责神经酰胺代谢。 该候选人试图阐明神经酰胺积累使 HSC 失活的机制，并定义 aCDase 在纤维化进展中的作用 该研究的具体目标是：1) 确定神经酰胺如何发挥作用。 介导 HSC 失活；2) 确定 aCDase 消耗和抑制对纤维化的影响 在第一个目标下，申请人将利用包括以下在内的技术组合。 免疫印迹、小分子抑制和激活、RNA 干扰以及组成型表达 激活突变蛋白以了解神经酰胺如何使 HSC 失活全基因组表达分析。 将进行以阐明神经酰胺的转录靶点 根据第二个目标，申请人将进行。 产生条件性成纤维细胞特异性 aCDase 缺失的小鼠，并将确定其程度 他们在两个独立的肝纤维化模型中经历了纤维化减少，她还将确定 aCDase 小分子抑制剂的体内抗纤维化作用作为该提案的一个组成部分， 候选人的职业发展将通过参与高级课程和研究得到补充 研讨会以发展分子生物学、全基因组转录分析和体内模型方面的专业知识 正式的指导委员会和咨询团队将提供监督、指导和指导。 协助候选人实现其目标的研究环境，其中包括 MGH GI 部门， 哈佛纤维化网络、哈佛大学学院、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，以确保候选人取得成功。 获奖者将通过为以下领域做出贡献而成为一名独立的基础科学研究者 了解鞘脂神经酰胺的作用及其对 HSC 体外和体内失活的作用。