Regulation of hepatic stellate cells in development and alcoholic liver injury

描述（由申请人提供）：这是加州大学弗朗西斯科分校博士后尹春跃博士的K99/R 00独立之路奖申请书。尹博士是酒精性肝病（ALD）研究领域的年轻研究者。该K99/R 00奖项将为Yin博士提供必要的支持，以实现以下目标：1）获得肝星状细胞（HSC）和酒精性肝损伤的专业知识; 2）开发研究斑马鱼HSC的新工具; 3）发展独立的研究生涯。为了实现这些目标，Yin博士组建了一个指导团队，由主要导师，UCSF生物化学和生物物理学教授Didier Mackier博士（斑马鱼肝脏发育专家）和共同导师，UCSF医学教授Jacquelyn Maher博士（酒精性肝损伤专家）组成。ALD是酒精相关发病率和死亡率的主要原因之一。肝星状细胞的激活是酒精性肝损伤的关键事件，但目前对肝星状细胞在酒精性肝损伤中的调控机制的认识还很有限。尹博士的长期目标是阐明酒精性肝损伤中HSC的细胞反应。本申请的总体目标是通过使用斑马鱼模型来了解HSC与邻近的窦状隙内皮细胞（SEC）在肝脏发育和急性酒精损伤中的相互作用。中心假设是HSC和SEC之间的旁分泌信号是HSC发育所必需的，并调节它们对酒精的反应。Yin博士将通过追求三个具体目标来实现该提案的目标：1）了解SEC在HSC发育中的作用; 2）确定HSC和SEC对急性酒精霍利奇性肝损伤的反应; 3）了解HSC-SEC相互作用对急性酒精霍利奇性肝损伤的分子基础。在目标1中，她假设在斑马鱼中HSC和SEC不共享共同的前体，但SEC对于HSC发育是必不可少的。她将通过谱系追踪实验和在发育过程中操纵HSC和SEC之间的相互作用来验证这一假设。在目标2中，Yin博士将通过实时成像实验来表征HSC和SEC对急性酒精暴露的细胞反应。在目标3中，她将进行基因谱分析，以表征HSC和SEC对急性酒精治疗反应的分子机制。她还将测试血小板衍生生长因子在调节HSC-SEC相互作用中的作用。这项研究具有创新性，因为它建立了一种新的斑马鱼模型，用于研究酒精性肝损伤中HSC与SEC的相互作用。这项研究也很有意义，因为它是一系列研究的第一步，有望阐明酒精性肝损伤中HSC活化的机制。拟议研究的基本原理是，对发育和急性酒精暴露中HSC的全面表征将为我们对ALD中HSC的理解提供新的见解，并可能转化为新的治疗靶点。 公共卫生相关性：拟议的研究与公共卫生有关，因为对肝星状细胞生物学的深入了解将使我们能够开发调节其激活的工具，这可能导致酒精性肝病的预防或逆转。因此，拟议中的研究与NIH的使命的一部分有关，该使命涉及发展有助于延长健康生命和减少疾病和残疾负担的基础知识。

DESCRIPTION (provided by applicant): This is an application for the K99/R00 Pathway to Independence Award for Dr. Chunyue Yin, a post- doctoral fellow at the University of California, San Francisco. Dr. Yin is establishing herself as a young investi- gator in the research of alcoholic liver disease (ALD). This K99/R00 award will provide Dr. Yin with the support necessary to accomplish the following goals: 1) to gain expertise in hepatic stellate cells (HSC) and alcoholic liver injury; 2) to develop new tools for studying HSC in zebrafish; and 3) to develop an independent research career. To achieve these goals, Dr. Yin has assembled a mentoring team comprised of a primary mentor, Dr. Didier Stainier, Professor of Biochemistry and Biophysics at UCSF, who is an expert in zebrafish liver development, and a co-mentor, Dr. Jacquelyn Maher, Professor of Medicine at UCSF, who is an expert in al- coholic liver injury. ALD is one of the leading causes of alcohol-related morbidity and mortality. Activation of HSC is the key event in ALD, but our understanding of the regulation of HSC in alcoholic liver injury is limited. Dr. Yin's long-term goal is to elucidate the cellular responses of HSC in alcoholic liver injury. The overall objective of this application is to understand the interactions between HSC and neighboring sinusoidal endothelial cells (SEC) in liver development and acute alcoholic injury by using the zebrafish model. The central hypothesis is that paracrine signals between HSC and SEC are required for HSC development and regulate their behaviors in response to alcohol. Dr. Yin will achieve the objective of the proposal by pursuing three specific aims: 1) Understand the roles of SEC in HSC development; 2) Determine the responses of HSC and SEC to acute al- coholic liver injury; and 3) Understand the molecular basis of HSC-SEC interactions in response to acute alco- holic liver injury. In Aim 1, she hypothesizes that in zebrafish HSC and SEC do not share a common precur- sor, yet SEC are essential for HSC development. She will test this hypothesis by lineage-tracing experiments and by manipulating the interactions between HSC and SEC during development. In Aim 2, Dr. Yin will char- acterize the cellular responses of HSC and SEC to acute alcohol exposure by time-lapse live imaging experi- ments. In Aim 3, she will perform gene-profiling analyses to characterize the molecular mechanisms underly- ing the responses of HSC and SEC to acute alcohol treatment. She will also test the role of Platelet-Derived Growth Factor in regulating HSC-SEC interactions. The proposed research is innovative because it estab- lishes a novel zebrafish model for studying HSC-SEC interactions in alcoholic liver injury. The proposed re- search is also significant because it is the first step in a continuum of research that is expected to elucidate the mechanisms of HSC activation in alcoholic liver injury. The rationale for the proposed research is that a com- prehensive characterization of HSC in development and acute alcohol exposure will provide novel insights into our understanding of HSC in ALD, and may translate into new targets for therapy. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because a thorough understanding of hepatic stellate cell biology will enable us to develop tools to modulate their activation, which may potentially lead to prevention or reversion of alcoholic liver disease. Thus, the proposed research is relevant to the part of NIH's mission that pertains to developing fundamental knowledge that will help to extend healthy life and reduce the burdens of illness and disability.