Regulation of hepatic stellate cells in development and alcoholic liver injury

描述(申请人提供)：这是为加州大学旧金山分校博士后尹春月博士申请K99/R00独立之路奖。殷博士在酒精性肝病(ALD)的研究中确立了自己的年轻研究者的地位。K99/R00奖项将为殷博士提供必要的支持，以实现以下目标：1)获得在肝星状细胞(HSC)和酒精性肝损伤方面的专业知识；2)开发研究斑马鱼HSC的新工具；以及3)发展独立的研究事业。为了实现这些目标，殷博士组建了一个导师团队，成员包括主要导师、加州大学旧金山分校生物化学和生物物理学教授、斑马鱼肝脏发育专家Didier Stainier博士和共同导师、加州大学旧金山分校医学教授、al-Coholic肝脏损伤专家Jacquelyn Maher博士。ALD是与酒精相关的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病的关键事件，但对肝星状细胞在酒精性肝损伤中的调节作用了解有限。殷博士的长期目标是阐明HSC在酒精性肝损伤中的细胞反应。这项应用的总体目标是通过斑马鱼模型了解HSC与邻近的肝窦内皮细胞(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，并调节其对酒精的反应行为。殷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发展中的作用；2)确定HSC和SEC对急性酒精性肝损伤的反应；以及3)了解HSC-SEC相互作用应对急性酒精性肝损伤的分子基础。在目标1中，她假设斑马鱼的HSC和SEC没有共同的前体，但SEC对HSC的发育是必不可少的。她将通过血统追踪实验和在发育过程中操纵HSC和SEC之间的相互作用来检验这一假说。在目标2中，殷博士将通过时间推移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，她将进行基因图谱分析，以表征HSC和SEC对急性酒精治疗反应不足的分子机制。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来研究HSC-SEC在酒精性肝损伤中的相互作用。这项拟议的研究也具有重要意义，因为这是一系列研究的第一步，有望阐明酒精性肝损伤中HSC激活的机制。这项拟议研究的基本原理是，全面描述HSC在发育和急性酒精暴露中的特征将为我们理解ALD中的HSC提供新的见解，并可能转化为新的治疗靶点。

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.