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

摘要 项目标题：肝细胞来源的MIF：酒精性肝病的关键因素 酒精性肝病(ALD)是全世界可预防的发病率和死亡率的重要原因。酒精- 在过去的几十年里，相关的发病率和死亡率保持不变，发达国家的饮酒率 国家数量继续增加。ALD是一种包括脂肪变性、酒精性肝炎(AH)、 肝硬变，甚至发展为肝癌。ALD的治疗方法，特别是AH，40年来没有变化 而且对相当比例的ALD患者无效。寻找ALD的新生物标志物 靶向治疗的进展和发展对公共卫生和福利具有显著的利益。 巨噬细胞迁移抑制因子(MIF)是一种普遍表达的天然免疫调节因子，与 趋化因子和细胞因子样活性与人类ALD和酒精喂养模型有关 动物。急性肝炎和肝硬变患者循环中MIF增加，MIF蛋白表达显著增加 在AH患者的肝细胞中，MIF是预测AH患者肝脏发病率和患者死亡率的指标。在动物身上 酒精喂养模型，MIF缺乏对酒精性肝损伤有保护作用，并使乙醇正常化。 诱导炎症和肝细胞内质网应激。综上所述，这些数据确定了肝细胞可能是 ALD中MIF异常表达和下游活性的组织来源和作用部位。这样做的目的是 建议对长期过量饮酒增加肝细胞MIF释放的假设进行检验 作为酒精性肝病进展的有效上游调节因子，通过控制 趋化因子表达、趋化因子包装与肝脏内质网应激。本文提出的工作将利用 组织特异性MIF基因敲除、MIF受体基因敲除和一种新的小分子MIF抑制剂的研究 肝细胞来源的MIF在多种酒精喂养动物模型中的作用 肝细胞培养，以及在人类ALD患者组织中。此外，MIF缺乏的保护性表型 小鼠与肝脏和循环细胞外循环中趋化因子表达的深刻变化有关 小泡(EV)。电动汽车是关键的细胞内通讯媒介，电动汽车中的趋化因子货物是 在乙醇和MIF依赖的机制中都非常动态。到那时，这项提案的另一个目标是 过渡，以探讨是什么导致ALD肝脏MIF表达增加。对数据库进行搜索 ALD患者肝脏的microcoRNAs(GEO系列GSE59492)显示，MIF的一个强大的负调控因子 在ALD的谱系中，miR-451的表达降低，并且对酒精中毒的病因具有选择性。 疾病。然后，该项目将扩大研究EVS作为ALD治疗转移的新机制，因为 拟议的研究将寻求通过恢复miR-451的表达以及 选择性地抑制肝脏中的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.