A PRECLINICAL MODEL OF ALCOHOLIC HEPATITIS

酒精性肝炎的临床前模型

• 批准号: 9792231
• 负责人: ARUN J SANYAL
• 金额: $ 19.34万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-25 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9792231
• 关键词: Acceleration; Alcohol Phenotype; Alcohol consumption; Alcohol withdrawal syndrome; Alcoholic Hepatitis; Alcoholic steatohepatitis; Alcohols; Animal Model; Bilirubin; Chronic; Data; Development; Diet; Disease; Disease Progression; Disease regression; Dose; Epigenetic Process; Fibrosis; Funding Mechanisms; Generations; Genes; Genetic Drift; Goals; High Fat Diet; Histology; Human; Inbreeding; Individual; Insulin Resistance; Knowledge; Liver; Malnutrition; Modeling; Morbidity - disease rate; Mouse Strains; Mus; Mutation; National Institute on Alcohol Abuse and Alcoholism; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Parents; Pathway interactions; Patients; Pattern; Phase; Pre-Clinical Model; Predisposition; Process; Protocols documentation; Public Health; Reagent; Research Personnel; Role; Safety; Sampling; Severities; Signal Pathway; Signal Transduction; Site; Steatohepatitis; Supervision; Testing; Therapeutic; Thinness; Translating; Validation; alcohol effect; alcohol exposure; base; clinically significant; dietary manipulation; drug testing; druggable target; feeding; high reward; high risk; human data; insight; liver function; liver injury; metabolome; microbiome; microbiome analysis; mortality; mouse model; mutant; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; obesogenic; offspring; prevent; response; therapeutic development; therapeutic target; transcriptome

Alcoholic Steatohepatitis (ASH) is a major cause of liver related mortality. Despite its public health significance, there has been limited therapeutic advances for ASH. A principal barrier to acceleration of therapeutics is the availability of a robust preclinical model of ASH which recapitulates human ASH. Such models are needed to better understand the role of specific genes and pathways in human ASH e.g. the PNPLA3 mutation which is associated with severe ASH and test promising compounds for both efficacy and safety. In this UH2/UH3 application, we propose to provide “proof of concept” that our recently validated diet-induced animal model of NAFLD (DIAMOND) can be leveraged as a model of ASH with specific alcohol feeding protocols and will test the following novel hypothesis: an inbred isogenic crossed C57Bl/6J and 129S1/SvlmJ (B6/S129) mouse strain will develop steatohepatitis that recapitulates the key features of human ASH with alcohol feeding. Furthermore, liver-specific expression of the human mutant I148M PNPLA3 in this mouse will accelerate the development of ASH upon alcohol feeding. As per RFA AA-18-006, the studies will be performed in two phases: UH2 Phase (yrs. 01-02): To demonstrate that alcohol feeding causes steatohepatitis resembling human ASH with respect to histology, markers of liver injury and function, and activation of signaling pathways in an isogenic strain of B6/S129 mice. Alcohol feeding will be done along with chow- or an obesogenic-diet. We will test the effect of the NIAAA alcohol model feeding strategy (to be performed at NIAAA under supervision by Dr. Gao) and an alternate strategy where we will perform single ascending dose (SAD) and multiple ascending dose (MAD) studies that will provide optimized alcohol feeding strategies including amounts given chronically along with binges to allow a phenotype of ASH to develop. We will also test the ability to accelerate development of ASH by liver-specific expression of the human I148M PNPLA3 mutant gene. These data are based on preliminary data indicating that NASH can be accelerated by this maneuver. A minimal requirement for development of ASH (steatohepatitis, increased AST and bilirubin) will be needed to proceed to the UH3 phase. UH3 Phase (yrs. 03-05): To further validate the model, define the course of disease progression and regression by modulation of alcohol intake, and the impact of ASH on the susceptibility to ASH in subsequent generations. The transcriptome, metabolome and microbiome of the model will be related to human data from the AlcHepNet consortium. Also, the effects of alcohol withdrawal after varying durations of exposure will be tested to define the “off-response”. We will also perform studies to determine the susceptibility of offspring of mice that have been allowed to develop ASH and then recover by withdrawing alcohol. Microbiome analyses will be done at the UCSD site of AlcHepNet. The investigators have the required expertise in ASH and mouse models of NASH and ASH. Together the studies will have a high impact by providing a mouse model of ASH. The project also meets the high-risk high reward criteria for the UH2/UH3 funding mechanism.

酒精性脂肪性肝炎（ASH）是肝脏相关死亡的主要原因。尽管它对公众健康很重要， 对于ASH的治疗进展有限。加速治疗的主要障碍是 一个强大的临床前模型的可用性，它概括了人类ASH。需要这些模型来 更好地了解特定基因和途径在人类ASH中的作用，例如PNPLA 3突变， 与严重ASH相关，并测试有希望的化合物的有效性和安全性。在此UH 2/UH 3 应用，我们建议提供“概念证明”，我们最近验证的饮食诱导的动物模型， NAFLD（DIAMOND）可用作具有特定酒精喂养方案的ASH模型，并将测试 以下新假设：近交系C57 B1/6 J和129 S1/SvlmJ（B6/S129）小鼠品系的同基因杂交 将发展脂肪性肝炎，其概括了人类酒精摄入ASH的关键特征。此外，委员会认为， 人突变体I148 M PNPLA 3在这只小鼠中的肝脏特异性表达将加速 酒精喂食后的灰烬。根据RFA AA-18-006，研究将分两个阶段进行：UH 2阶段 (yrs. 01-02）：为了证明酒精喂养导致类似于人类ASH的脂肪性肝炎， 组织学，肝损伤和功能的标志物，以及信号通路的激活，在一个同基因株的 B6/S129小鼠。酒精喂养将沿着食物或致肥胖饮食。我们将测试 NIAAA酒精模型喂养策略（将在高博士的监督下在NIAAA进行）和 我们将进行单次给药剂量递增（SAD）和多次给药剂量递增（MAD）的替代策略 研究将提供优化的酒精喂养策略，包括长期沿着 导致ASH的表型形成。我们还将测试加速ASH开发的能力 通过人I148 M PNPLA 3突变基因的肝脏特异性表达。这些数据是根据初步数据得出的 这表明NASH可以通过这种方法加速。ASH开发的最低要求 （脂肪性肝炎，AST和胆红素升高）将需要继续进行UH 3阶段。UH 3阶段（年 03-05）：为了进一步验证该模型，通过调节 酒精摄入量，以及ASH对后代对ASH易感性的影响。的 该模型的转录组、代谢组和微生物组将与来自AlcHepNet的人类数据相关 财团此外，将测试不同暴露时间后戒酒的影响，以确定 “关闭响应”。我们还将进行研究，以确定小鼠的后代的易感性， 被允许出现灰烬，然后通过戒酒来恢复。微生物组分析将在 AlcHepNet的UCSD网站。研究者在ASH和NASH小鼠模型方面具有所需的专业知识 还有灰烬这些研究将通过提供ASH的小鼠模型产生很大的影响。该项目还 符合UH 2/UH 3供资机制的高风险高回报标准。