Role of RIPK1 and RIPK3 in liver injury.

RIPK1 和 RIPK3 在肝损伤中的作用。

基本信息

批准号：
10445150
负责人：
Lily Dara
金额：
$ 8.54万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2022-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10445150
关键词：
Acetaminophen Acute Acute Liver Failure Advisory Committees Albumins Apoptosis Area Award California Caspase Cell Death Cell Death Signaling Process Cell Survival Cell membrane Cells Cessation of life Chronic Clinical Collaborations Critiques Data Development Plans Education Endothelial Cells Endothelium Ensure Environment Equipment Experimental Models Extinction (Psychology)Fibrosis Funding Goals Hemorrhage Hepatic Hepatocyte Hepatotoxicity In Vitro Inflammation Injury Interruption Intervention K-Series Research Career Programs Knock-out Lead Lilium Liver Liver Failure Liver diseases Longitudinal Studies LoxP-flanked allele MAP Kinase Gene MAPK8 gene Mediating Mediator of activation protein Mentored Clinical Scientist Development Award (K08)Mentorship Modeling Molecular Mus N-acetyl-4-benzoquinoneimine National Institute of Diabetes and Digestive and Kidney Diseases Necrosis Operative Surgical Procedures Outcome Pathway interactions Pharmacology Phosphotransferases Physicians Predisposition Protein Biochemistry Protein Kinase Proteins RIPK1 gene RIPK3 gene Regulation Reperfusion Injury Research Resources Role Rupture Scientist Signal Pathway Signal Transduction Steatohepatitis Structure TNF gene TRAF2 gene Toxic effect Training Transgenic Mice Transgenic Organisms United States Universities Work acute liver injury base brain cell cadherin 5 career career development cell type chronic liver disease chronic liver injury clinical application clinically relevant endoplasmic reticulum stress experimental study genetic manipulation heart cell in vivo in vivo Model kidney cell knock-down knockout animal liver injury liver ischemia mouse model multidisciplinary mutant new therapeutic target non-alcoholic fatty liver disease novel problem drinker programs protein expression protein function public health relevance response scaffold therapeutic target

项目摘要

DESCRIPTION (provided by applicant): The Receptor Interacting Protein Kinases (RIPK1 and RIPK3) are proteins involved in cell death and survival. The role of RIPK1 in regulating these outcomes is determined by context and the kinase versus scaffolding function of the protein. Necroptosis is a form of regulated cell death dependent on the kinase function of RIPK1, activating the RIPK1-RIPK3 necrosome leading to the activation of MLKL and cell membrane rupture when caspases are inhibited. Using the acetaminophen (APAP) liver injury model, we found no role for necroptosis as RIPK3-/- and MLKL -/- mice are not protected. However, we identified a novel scaffolding function for RIPK1 in APAP toxicity. We have shown that knocking down RIPK1 protects mice from APAP hepatotoxicity upstream of JNK, a key mediator of APAP induced cell death. Interestingly, our preliminary studies reveal RIPK1 kinase-dead transgenic mice are not protected from APAP, strongly suggesting a scaffolding function of the protein in cell death signaling. In addition, we have found a cell specific distribution for hepatic RIPK3 protein, with abundant protein expression in the non-parenchymal cells, including liver sinusoidal endothelial cells (LSECs). These findings have lead to the following overarching hypothesis, which forms the basis of this career development proposal in two clinically relevant liver injury models: APAP hepatotoxicity, and ischemia reperfusion injury (IRI). RIPK1 regulates cell death in the liver through a scaffolding function in hepatocytes and a kinase function in LSECs. For the first two years of the award we will focus on aim1, which will systematically explore the scaffolding function of RIPK1 in hepatocytes. We will also determine the interacting proteins with RIPK1 and the role of RIPK1 in the signaling pathway that leads to JNK activation in APAP toxicity and other ER stress models. Subsequently, in the second aim of the proposal we will explore the effect of cell type specific deletion of RIPK1 and RIPK3 in models of liver injury by examining liver injury in mice with conditional hepatocyte or endothelial deletions. The proposed studies are the core components of the Mentored Clinical Scientist Development Award (K08) for Dr. Lily Dara, around which a structured program for Dr. Dara has been built. Dr. Dara has identified four target areas for additional training throughout the K Award period: protein biochemistry and signaling, transgenic mice models, LSEC pathobiology, and the liver ischemia reperfusion injury model. To achieve these goals, she has devised a 5-year career development plan and assembled a multidisciplinary advisory team of scientists specializing in each of these core components to guide her work and critique her findings. The University of Southern California has a long-standing tradition in excellence in liver research. The NIDDK- funded USC Research Center for Liver Disease is an excellent resource for the candidate, providing both practical facilities and equipment through its cores, and the opportunity for scientific exchange and collaboration through its large membership base. The immediate focus of this project in the next five years is the role of RIPK1 and RIPK3 in models of acute liver injury. After completion of these studies the long-term goal of Dr. Dara is to extend these findings to experimental models of chronic liver injury such as alcoholic and nonalcoholic fatty liver disease/steatohepatitis. The experiments planned in this proposal, the guidance of the mentorship advisory committee, and the ideal training environment at USC will ensure Dr. Dara's successful transition to an independent physician-scientist while advancing the understanding of the role of RIP kinases in liver disease.

描述（由申请人提供）：受体相互作用的蛋白激酶（RIPK1和RIPK3）是参与细胞死亡和存活的蛋白质。 RIPK1在调节这些结果中的作用取决于上下文以及蛋白质的激酶与脚手架功能。坏死性是根据RIPK1的激酶功能的调节细胞死亡形式，激活RIPK1-RIPK3坏死体，导致MLKL和细胞膜破裂的激活时，当caspase被抑制时。使用对乙酰氨基酚（APAP）肝损伤模型，我们没有发现坏死作用，因为RIPK3 - / - 和MLKL - / - 小鼠不受保护。但是，我们确定了RIPK1在APAP毒性中的新型脚手架功能。我们已经表明，击倒RIPK1可保护小鼠免受JNK上游的APAP肝毒性，这是APAP诱导细胞死亡的关键介体。有趣的是，我们的初步研究表明，RIPK1激酶已故转基因小鼠不受APAP的保护，强烈表明该蛋白质在细胞死亡信号传导中具有脚手架功能。此外，我们发现了肝RIPK3蛋白的细胞特异性分布，在非核苷细胞中具有丰富的蛋白质表达，包括肝脏正义性内皮细胞（LSEC）。这些发现导致了以下基本假设，这是两个临床相关的肝损伤模型中该职业发展建议的基础：APAP肝毒性和缺血再灌注损伤（IRI）。 RIPK1通过肝细胞中的脚手架功能和LSEC中的激酶功能调节肝脏中的细胞死亡。在奖项的头两年中，我们将重点关注AIM1，该AIM1将系统地探索RIPK1在肝细胞中的脚手架功能。我们还将确定与RIPK1的相互作用蛋白以及RIPK1在信号通路中的作用，从而导致APAP毒性和其他ER应力模型中JNK激活。随后，在提案的第二个目标中，我们将通过检查有条件性肝细胞或内皮缺失的小鼠肝损伤的肝损伤模型中RIPK1和RIPK3的细胞类型特异性缺失的影响。拟议的研究是莉莉·达拉（Lily Dara）博士的指导临床科学家发展奖（K08）的核心组成部分，围绕该博士为Dara博士建立了结构化计划。 Dara博士在整个K颁发期间确定了四个目标领域，以进行额外的培训：蛋白质生物化学和信号传导，转基因小鼠模型，LSEC病理生物学和肝缺血再灌注损伤模型。为了实现这些目标，她制定了一项为期5年的职业发展计划，并组建了一个专门研究这些核心组成部分的科学家的多学科咨询团队，以指导她的工作和批评她的发现。南加州大学在肝脏研究方面具有悠久的卓越传统。 NIDDK资助的USC肝病研究中心是候选人的绝佳资源，通过其核心提供实用的设施和设备，以及通过其大型会员基础进行科学交流和协作的机会。在未来五年中，该项目的直接重点是RIPK1和RIPK3在急性肝损伤模型中的作用。完成这些研究后，DARA博士的长期目标是将这些发现扩展到慢性肝损伤的实验模型，例如酒精和非酒精性脂肪肝病/脂肪性肝炎。在本提案中计划的实验，指导咨询委员会的指导以及USC的理想培训环境将确保Dara博士的成功过渡到独立的医生 - 科学家，同时促进对RIP激酶在肝病中的作用的理解。