The Role of miR-122 in Alcoholic Liver Disease

miR-122 在酒精性肝病中的作用

• 批准号: 9097481
• 负责人: Abhishek Satishchandran
• 金额: $ 3.47万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-17 至 2018-07-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9097481
• 关键词: Abstinence; Acetaminophen; Alcohol abuse; Alcohol consumption; Alcoholic Liver Diseases; Alcoholic liver damage; Alcohols; Biochemical Process; Bioinformatics; Biological; Biological Assay; C57BL/6 Mouse; Carrier Proteins; Cell Cycle; Cell Proliferation; Cell physiology; Chronic; Cirrhosis; Cyclin D1; Data; Dependovirus; Development; Diagnosis; Diet; Disease; Doctor of Medicine; Doctor of Philosophy; Equilibrium; Feedback; Fellowship; Fibrosis; Funding; HNF4A gene; Health; Hepatic; Hepatitis; Hepatocyte; Histologic; Homeostasis; Inflammation; Inflammatory; Knowledge; Laboratories; Lipids; Liver; Liver Cirrhosis; Liver diseases; Mediator of activation protein; Metabolism; MicroRNAs; Modality; Modeling; Molecular; Morbidity - disease rate; Mus; Natural regeneration; Neoplasm Metastasis; Nucleotides; Pathogenesis; Patients; Physiological Processes; Prevention; Primary carcinoma of the liver cells; Process; Publications; RNA; Recombinant adeno-associated virus (rAAV); Recovery; Regulation; Regulator Genes; Role; Serotyping; Serum; Severities; Signal Pathway; Signal Transduction; Steatohepatitis; System; Therapeutic; Toxin; Training; Work; burden of illness; carcinogenesis; chronic alcohol ingestion; cofactor; cytokine; feeding; gene function; gene therapy; hepatocyte nuclear factor; hypoxia inducible factor 1; in vivo; knock-down; liver injury; liver metabolism; mortality; mouse model; outcome forecast; overexpression; prevent; problem drinker; research study; small hairpin RNA; targeted delivery; tool; transcription factor; vector

DESCRIPTION (provided by applicant): Alcohol abuse has been attributed to 3.2% of the world's disease burden. Chronic abuse manifests as reversible steatosis, steatohepatitis and/or irreversible cirrhosis. Regardless abstinence, 5-15% of patients with steatohepatitis still progress to cirrhosis and hepatocellular carcinoma (HCC). Studies have demonstrated the role of miR-122 as a mediator of hepatic metabolism, cellular differentiation and the development of HCC. Our laboratory has shown decreased miR-122 in a four-week chronic-alcohol mouse model. The role of miR-122 in ALD is unknown. Bioinformatic miRNA target prediction tools suggest Hypoxia-Inducible Factor 1-� (HIF1�) is a primary target of miR-122. HIF1� is critical to the progression of ALD. Specific Aim 1 will study the role of miR-122 in the pathogenesis of chronic alcohol-induced hepatitis. C57Bl/6 mice will be transfected using a hepatocyte-tropic adeno-associated virus serotype 8 (rAAV) vector to either knockdown or overexpress miR-122 via tough decoy (TuD) or the miR-122 respectively. Mice will be maintained on Lieber-DeCarli diet for 28 days. On day 28 mice will be withdrawn from alcohol and given 150- mg/kg acetaminophen (APAP). Mice will be sacrificed on day 30 to assess regeneration. Livers sections will be analyzed histologically for morphological changes, lipid accumulation, and regeneration. In addition, we will determine the impact on miR-122 modulation on hepatic inflammation and differentiation through expression analysis of inflammatory cytokines and cell cycle regulators associated with ALD. MiR-122 has been correlated with a network of Liver Enriched Transcription Factors (LETFs). Collectively, these LETFs function as master regulators of hepatic function. HNF6�, specifically, has been shown to function in a positive feedback loop with miR-122. Specific Aim 2 will examine the effect of miR-122, its overexpression and knockdown, on HNF6� in a chronic-alcohol model. We will also examine if miR-122 can reduce ALD through enhancement of HNF6� and the LETF network. The rAAV8 vector stated above will deliver anti- HNF6� shRNA or rHNF6 in vivo. The mice will be treated and assayed as described above. During the first year of this fellowship we aim to examine the effect of miR-122 on the severity of ALD development using gene therapy. The remaining two years of my Ph.D. (funding yrs 2 & 3) will be spent studying the effect miR-122 and HNF6� regulation of the LETF network in the progression of ALD. During years 4 and 5 of funding I shall complete my M.D. training while finalizing any work required for publication. Collectively, we propose to examine two potential avenues by which miR-122 may serve as a potential therapeutic modality. First, is the development of steatosis though inhibition of HIF1� and secondly, is the regeneration after alcohol and APAP-induced liver injury.

描述（由申请人提供）：酒精滥用已被认为是世界疾病负担的3.2%。慢性滥用表现为可逆性脂肪变性、脂肪性肝炎和/或不可逆性肝硬化。无论是否戒酒，仍有5-15%的脂肪性肝炎患者进展为肝硬化和肝细胞癌（HCC）。研究表明，miR-122在肝脏代谢、细胞分化和HCC发生中的作用。我们的实验室在四周的慢性酒精小鼠模型中显示miR-122降低。miR-122在ALD中的作用尚不清楚。生物信息学miRNA靶标预测工具提示缺氧诱导因子1- α (HIF1 α)是miR-122的主要靶标。HIF1对ALD的进展至关重要。特异性目的1将研究miR-122在慢性酒精性肝炎发病机制中的作用。使用嗜肝细胞腺相关病毒血清型8 （rAAV）载体转染C57Bl/6小鼠，分别通过tough诱饵（TuD）或miR-122敲低或过表达miR-122。饲喂Lieber-DeCarli日粮28 d。第28天，小鼠停止饮酒，给予对乙酰氨基酚（APAP） 150 mg/kg。第30天处死小鼠，评估再生情况。肝脏切片将在组织学上分析形态学变化，脂质积累和再生。此外，我们将通过分析与ALD相关的炎症细胞因子和细胞周期调节因子的表达来确定miR-122调节对肝脏炎症和分化的影响。MiR-122与肝脏富集转录因子（LETFs）网络相关。总的来说，这些letf是肝功能的主要调节者。具体来说，HNF6 -已被证明与miR-122在一个正反馈回路中起作用。特异性目的2将在慢性酒精模型中检测miR-122及其过表达和敲低对HNF6的影响。我们还将研究miR-122是否可以通过增强HNF6 α和LETF网络来减少ALD。上述rAAV8载体将在体内传递抗HNF6 shRNA或rHNF6。小鼠将按照上述方法进行处理和分析。在这项研究的第一年，我们的目标是通过基因治疗检查miR-122对ALD发展严重程度的影响。博士剩余的两年（资助第2年和第3年）将用于研究miR-122和HNF6调控LETF网络在ALD进展中的作用。在资助的第四年和第五年，我将完成我的医学博士培训，同时完成出版所需的任何工作。总之，我们建议研究miR-122可能作为潜在治疗方式的两种潜在途径。首先是通过抑制HIF1 -而发生脂肪变性，其次是酒精和apap诱导的肝损伤后的肝再生。