Monocyte regulation in human alcoholic hepatitis.

人类酒精性肝炎中的单核细胞调节。

基本信息

批准号：
9373415
负责人：
STEVEN A WEINMAN
金额：
$ 21.99万
依托单位：
UNIVERSITY OF KANSAS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9373415
关键词：
Abnormal Monocyte Acetylation Acute Acute Alcoholic Hepatitis Alcohol consumption Alcoholic Hepatitis Alcoholic Liver Diseases Alcoholic beverage heavy drinker Alcohols Anti-Inflammatory Agents Anti-inflammatory Apoptosis Apoptotic Autopsy Binding Cell Culture Techniques Cessation of life Data Deacetylase Defect Development Disease FOXO3A gene Failure Future Goals Heavy Drinking Hepatic Human In Vitro Individual Inflammation Inflammatory Inflammatory Response Intervention Kupffer Cells Lead Liver Liver Failure Liver diseases MAPK8 gene Minority Modification Mus Nature Pathogenesis Pathway interactions Patients Penetrance Peripheral Blood Mononuclear Cell Phenotype Phosphorylation Play Predisposition Production Protein Isoforms Proteins Regulation Risk Role Sepsis Series Serine Specificity Specimen Steatohepatitis Stimulus System Testing Therapeutic Time Up-Regulation Variant Work acute liver disease alcohol response binge drinking cytokine disease phenotype experimental study human disease human subject immunoregulation individual patient insight liver injury macrophage monocyte mortality mouse model novel novel therapeutic intervention prevent problem drinker promoter response therapeutic target transcription factor

项目摘要

PROJECT DESCRIPTION/SUMMARY Acute alcoholic hepatitis (AH) is a severe inflammatory liver disease triggered by binge drinking. It has high mortality and treatment options are only poorly effective. AH only occurs in a small minority of heavy drinkers and most individuals appear to be protected from this form of liver injury. Previous work from our lab has identified the transcription factor FOXO3 as a possible protection factor and more recently we have determined that circulating monocytes from patients with AH have lost the ability to phosphorylate FOXO3 at S574 thus failing to generate a form of the protein (pFOXO3) that is normally induced by alcohol and appears to have protective, anti-inflammatory effects. This defect appears to result from abnormally high expression of the deacetylase SIRT7. It has long been known that macrophages and monocytes play an important role in the pathogenesis of AH. Blood monocyte counts are elevated in AH and both monocytes and macrophages have elevated inflammatory cytokine production in response to stimuli. This application will specifically examine blood monocytes and liver sections from patients with AH and heavy alcohol-consuming individuals who have never developed liver disease to determine the mechanisms that lead to abnormal FOXO3 phosphorylation in monocytes, the consequences of phosphorylation defects to abnormal monocyte function in AH, and whether or not abnormalities in the FOXO3 pathway serve as a susceptibility factor for the development of AH. We seek to test the hypothesis that alcohol consumption induces the formation of pFOXO3 in both monocytes and macrophages and pFOXO3 subsequently induces apoptosis and suppresses inflammatory cytokine production thus limiting alcohol-induced inflammation. We further hypothesize that high monocyte expression of SIRT7 prevents pFOXO3 formation and could serve as a potential therapeutic target. Finally, we propose to test whether variable ability to form pFOXO3 as a result of variations of SIRT7 expression plays a role in determining the intrinsic susceptibility of different individuals to alcoholic liver disease. We will test these hypotheses with two specific aims. Aim 1 will determine the relationship between pFOXO3, SIRT7 and inflammatory responses in circulating monocytes from patients with acute alcoholic hepatitis. These experiments will involve obtaining blood monocytes from patients with alcoholic hepatitis and appropriate controls, and examining the relationship between FOXO3 phosphorylation and in vitro apoptosis and cytokine responses. Aim 2 will examine the role of pFOXO3 in susceptibility to AH. These will examine blood monocytes from alcohol-consuming non-acutely ill patients with varying risk of development of AH, and liver sections from autopsy specimens from patients who died from alcohol-related MVAs. Overall, this proposal will use human disease specimens to rigorously test a novel mechanistic hypothesis of the pathogenesis of alcoholic hepatitis. and will extend the work from cell culture to mouse models to human patients with the ultimate goal of developing novel therapeutic approaches for this disease.

项目描述/摘要急性酒精性肝炎（AH）是一种由酗酒引发的严重炎症性肝病。它具有高死亡率和治疗方案效果不佳。 AH 仅发生在少数酗酒者中大多数人似乎都免受这种形式的肝损伤。我们实验室之前的工作有确定转录因子 FOXO3 作为可能的保护因子，最近我们确定 AH 患者的循环单核细胞失去了在 S574 位点磷酸化 FOXO3 的能力，因此未能生成通常由酒精诱导的蛋白质（pFOXO3）形式，并且似乎具有保护、抗炎作用。该缺陷似乎是由于异常高表达的脱乙酰酶 SIRT7。人们早就知道巨噬细胞和单核细胞在 AH 的发病机制。 AH 中血单核细胞计数升高，单核细胞和巨噬细胞均增加响应刺激而增加炎症细胞因子的产生。该应用程序将专门检查 AH 患者和酗酒者的血单核细胞和肝切片从未患过肝病以确定导致 FOXO3 磷酸化异常的机制单核细胞，磷酸化缺陷对 AH 中单核细胞功能异常的影响，以及是否 FOXO3 通路的异常或不异常是 AH 发生的易感因素。我们试图检验以下假设：饮酒会诱导单核细胞和单核细胞中 pFOXO3 的形成巨噬细胞和 pFOXO3 随后诱导细胞凋亡并抑制炎症细胞因子的产生从而限制酒精引起的炎症。我们进一步假设 SIRT7 的单核细胞高表达阻止 pFOXO3 的形成并可作为潜在的治疗靶点。最后，我们建议测试一下由于 SIRT7 表达的变化而形成 pFOXO3 的可变能力是否在确定不同个体对酒精性肝病的内在易感性。我们将测试这些具有两个特定目标的假设。目标 1 将确定 pFOXO3、SIRT7 和急性酒精性肝炎患者循环单核细胞的炎症反应。这些实验将涉及从酒精性肝炎患者身上获取血液单核细胞和适当的对照，并检查 FOXO3 磷酸化与体外细胞凋亡和细胞因子之间的关系回应。目标 2 将检查 pFOXO3 在 AH 易感性中的作用。这些将检查血液来自具有不同 AH 和肝脏发展风险的饮酒非急性疾病患者的单核细胞死于酒精相关 MVA 的患者尸检标本的切片。总体而言，该提案将使用人类疾病标本严格测试发病机制的新机制假设酒精性肝炎。并将把工作从细胞培养扩展到小鼠模型，再到患有该疾病的人类患者最终目标是开发针对这种疾病的新治疗方法。