Hepatocyte-derived MIF: a key contributor to Alcoholic Liver Disease

摘要 项目名称：肝细胞衍生的MIF：酒精性肝病的关键因素 酒精性肝病（ALD）是全球可预防的发病率和死亡率的重要原因。酒精- 相关的发病率和死亡率在过去几十年中保持不变，发达国家的饮酒率 国家继续增加。ALD是一种谱系障碍，包括脂肪变性、酒精性肝炎（AH）， 肝硬化甚至发展为肝癌。ALD的治疗，特别是AH，在40年内没有变化。 年以及无效的一个显着比例的ALD患者。识别ALD的新生物标志物 靶向治疗的进展和发展对公共健康和福利具有显著的意义。 巨噬细胞移动抑制因子（MIF），一种广泛表达的先天性免疫调节因子， 趋化因子和类胡萝卜素活性，与人类和酒精喂养模型中的ALD相关。 动物在AH和肝硬化患者中，MIF在循环中增加， 在AH患者的肝细胞中，MIF可预测AH的肝脏发病率和患者死亡率。在动物 模型的乙醇喂养，MIF缺乏保护乙醇诱导的肝损伤和正常化乙醇- 诱导炎症和肝细胞ER应激。总之，这些数据确定肝细胞可能是 异常MIF表达和ALD下游活性的组织来源和作用部位。的目的 这项研究的目的是检验慢性过量酒精增加肝细胞MIF释放的假设， 作为酒精性肝病进展的有效上游调节剂，通过控制 趋化因子表达、趋化因子包装和肝脏ER应激。本文提出的工作将利用 组织特异性MIF敲除、MIF受体敲除和一种新型小分子MIF抑制剂， 肝细胞衍生的MIF在动物多种酒精喂养模型中的作用， 肝细胞培养物和人ALD患者组织中。此外，MIF缺陷型中的保护性表型 小鼠与肝脏和循环细胞外基质中趋化因子表达的深刻变化有关。 囊泡（EV）。EV是关键的细胞内通讯介质，并且EV中的趋化因子货物是 在乙醇和MIF依赖性机制中都非常活跃。这项建议的另一个目的是， 过渡，以探索导致ALD中肝脏MIF表达增加的原因。数据库搜索 来自ALD患者肝脏的microRNA（GEO系列GSE 59492）揭示了MIF的有效负调节因子 miR-451的表达沿着ALD谱降低，并且对酒精性病因具有选择性， 疾病然后，该项目将扩展到研究EV作为ALD治疗转移的新机制， 提出的研究将通过恢复miR-451表达以及通过增加miR-451的表达来使MIF表达正常化。 选择性抑制肝脏中的MIF。

ABSTRACT Project Title: Hepatocyte-derived MIF: a key contributor to Alcoholic Liver Disease Alcoholic liver disease (ALD) is a significant cause of preventable morbidity and mortality worldwide. Alcohol- related morbidity and mortality has remained constant over the past decades, and drinking rates in developed countries continue to increase. ALD is a spectrum disorder encompassing steatosis, alcoholic hepatitis (AH), cirrhosis and even progressing to liver cancer. Therapies for ALD, and specifically AH, are unchanged for 40 years as well as ineffective for a significant proportion of ALD patients. Identifying novel biomarkers of ALD progression and the development of targeted therapies is of marked interest to public health and welfare. Macrophage Migration Inhibitory Factor (MIF), a ubiquitously expressed regulator of innate immunity with chemokine- and cytokine-like activities, is associated with ALD in humans and in ethanol feeding models in animals. MIF is increased in circulation in AH and cirrhosis patients, MIF protein expression is robustly increased in hepatocytes of AH patients, and MIF is predictive of liver morbidity and patient mortality in AH. In animal models of ethanol feeding, MIF deficiency protects from ethanol-induced liver injury and normalizes ethanol- induced inflammation and hepatocyte ER stress. Taken together, these data identified the hepatocyte as a likely tissue source and site of action for aberrant MIF expression and downstream activity in ALD. The aim of this proposal is to test the hypothesis that that chronic, excessive alcohol increases hepatocyte MIF release that acts as a potent upstream regulator of Alcoholic Liver Disease progression through control of chemokine expression, chemokine packaging, and liver ER stress. The work proposed herein will utilize tissue-specific MIF knockouts, MIF receptor knockouts and a novel small-molecule MIF inhibitor to interrogate the role of hepatocyte-derived MIF in multiple alcohol feeding models in animals, ethanol exposure in primary hepatocyte cultures, and in human ALD patient tissues. Furthermore, the protective phenotype in MIF-deficient mice is associated with profound changes in chemokine expression in the liver and in circulating extracellular vesicles (EV). EVs are pivotal intracellular communication mediators, and the chemokine cargoes in EVs are quite dynamic in both ethanol- and MIF-dependent mechanisms. The other aim of this proposal will then transition to explore what leads to this increased hepatic MIF expression in ALD. A database search of mircoRNAs from ALD patient livers (GEO series GSE59492) revealed that a potent negative regulator of MIF expression, miR-451, is decreased along the spectrum of ALD and was selective to an alcoholic etiology of disease. The project will then expand to study EVs as a novel mechanism of therapeutic transfer in ALD, as the proposed studies will look to normalize MIF expression through restoring miR-451 expression as well as selectively inhibit MIF in the liver.