The Role of miR-122 in Alcoholic Liver Disease

miR-122 在酒精性肝病中的作用

• 批准号: 8527387
• 负责人: Abhishek Satishchandran
• 金额: $ 2.87万
• 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/8527387
• 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; 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; 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; liver injury; mortality; mouse model; outcome forecast; overexpression; prevent; problem drinker; public health relevance; 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）：酒精滥用占世界疾病负担的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。无论是否禁欲，5-15%的脂肪性肝炎患者仍进展为肝硬化和肝细胞癌（HCC）。研究已经证明miR-122作为肝代谢、细胞分化和HCC发展的介导剂的作用。我们的实验室在一个为期四周的慢性酒精小鼠模型中显示了miR-122的减少。miR-122在ALD中的作用尚不清楚。生物信息学miRNA靶点预测工具表明缺氧诱导因子1-（HIF 1）是miR-122的主要靶点。HIF 1对ALD的进展至关重要。具体目标1将研究miR-122在慢性酒精性肝炎发病机制中的作用。将使用嗜肝细胞腺相关病毒血清型8（rAAV）载体转染C57 B1/6小鼠，以分别通过坚韧诱饵（TuD）敲低或过表达miR-122或miR-122。小鼠将维持Lieber-DeCarli饮食28天。在第28天，将小鼠从酒精中戒断并给予150- mg/kg对乙酰氨基酚（APAP）。将在第30天处死小鼠以评估再生。将对肝脏切片的形态学变化、脂质蓄积和再生进行组织学分析。此外，我们将通过与ALD相关的炎性细胞因子和细胞周期调节因子的表达分析来确定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（LETF）网络相关。总的来说，这些LETF作为肝功能的主要调节剂发挥作用。具体来说，HNF 6已被证明在与miR-122的正反馈回路中发挥作用。Specific Aim 2将在慢性酒精模型中检查miR-122，其过表达和敲低对HNF 6的影响。我们还将研究miR-122是否可以通过增强HNF 6和LETF网络来减少ALD。上述rAAV 8载体将在体内递送抗HNF 6 shRNA或rHNF 6。将如上所述处理和测定小鼠。在该研究的第一年，我们的目标是使用基因治疗来检查miR-122对ALD发展严重程度的影响。我博士学位剩下的两年（资助第2和第3年）将用于研究miR-122和HNF 6调节LETF网络在ALD进展中的作用。在第四年和第五年的资助中，我将完成我的医学博士学位。培训，同时完成出版所需的任何工作。总的来说，我们建议研究两种潜在的途径，通过这两种途径，miR-122可以作为一种潜在的治疗方式。首先，是通过抑制HIF 1 <$而发生脂肪变性，其次是酒精和APAP诱导的肝损伤后的再生。