The role of MKP-1/MAPK in hepatocytes and macrophages in alcohol-associated liver disease pathogenesis

MKP-1/MAPK在肝细胞和巨噬细胞中在酒精相关性肝病发病机制中的作用

• 批准号: 10679716
• 负责人: Mary Nancy Walter
• 金额: $ 3.65万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679716
• 关键词: Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcoholic beverage heavy drinker; Alcohols; Apoptotic; Area; Basic Science; Cell Count; Cell Line; Cell model; Cells; Cessation of life; Chronic; Cirrhosis; Clinical; Data; Development; Disease; Disease Progression; Down-Regulation; Endotoxins; Ethanol; FDA approved; Fostering; Galactosamine; Goals; Health; Hepatic; Hepatitis; Hepatocyte; Immune; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Injury; Intestinal permeability; Investigation; Knockout Mice; Kupffer Cells; Laboratories; Learning; Liver; Liver diseases; LoxP-flanked allele; MAPK1 gene; MAPK8 gene; Macrophage; Mediating; Medical; Mentors; Mitogen-Activated Protein Kinases; Modeling; Morbidity - disease rate; Mus; Myeloid Cells; N-terminal; National Institute on Alcohol Abuse and Alcoholism; Organ; Pathogenesis; Pathology; Phenotype; Phosphoric Monoester Hydrolases; Phosphotransferases; Play; Portal vein structure; Predisposition; Primary carcinoma of the liver cells; Production; Research; Research Personnel; Role; Scientist; Severity of illness; Signal Transduction; Specificity; Steatohepatitis; Supervision; TNF gene; Techniques; Testing; Tissues; Toxic effect; Training; Work; Writing; alcohol effect; alcohol exposure; career; career networking; chemokine; cytokine; experience; hepatocyte injury; in vivo; intrahepatic; liver inflammation; liver injury; member; microbial products; mortality; mouse model; new therapeutic target; novel; novel therapeutics; oral communication; p38 Mitogen Activated Protein Kinase; phosphatase-1 kinase; response; simple steatosis; skills; western diet

Project Summary Alcohol-associated liver disease (ALD) is responsible for nearly half of all deaths from liver disease. Despite the severity of this disease, there remain no FDA-approved treatments for any stage of the disease, highlighting the critical need to elucidate the underlying mechanisms of ALD progression to develop new therapies. Mitogen-activated protein kinase phosphatase 1 (MKP-1) is the archetypal member of the dual- specificity phosphatases with a pivotal role in organ inflammation and injury, including in the liver. Earlier work showed that decreased MKP-1 levels are associated with the activation of c-Jun N-terminal kinase (JNK), which triggers an apoptotic cascade leading to alcohol induced liver injury. However, the exact mechanisms of how MKP-1 downregulation leads to hepatocyte injury and/or inflammation in ALD are known. Moreover, the role of MKP-1 in macrophages and MAPK-driven inflammation in ALD has never been examined. Our preliminary studies show that MKP-1 deletion in hepatocytes augments alcohol-induced liver injury is a NIAAA chronic plus binge model of ALD. Data also suggest that MKP-1 deletion in hepatocytes sensitizes them to TNFα induced toxicity. As an aspiring scientist whose goal is to pursue a career in alcohol-related multi-organ pathology, investigating new players in ALD pathogenesis would not only help fill this medical need but also serve as an excellent platform from which to learn how to become a successful scientist. With the help and supervision of my mentors Dr. Gobejishvili (an expert in basic research on ALD) and Dr. McClain (an expert on clinical aspects of the disease) as well as several knowledgeable collaborators, I will investigate the role of alcohol-associated downregulation of MKP-1 in the pathogenesis of ALD. The first aim of this study is to further characterize the role of MKP-1 deletion in hepatocytes using hepatocyte specific MKP-1 knpockout mice in two different mouse models of ALD. I will isolate hepatocytes and immune cells from these models, examine the effect of MKP-1 deletion on pro-inflammatory chemokine production and immune cell phenotype and describe changes in hepatocyte and immune cell health and function. The second aim will determine the role of MKP-1 specifically in macrophages using RAW 264.7 macrophage cell line, primary Kupffer cells and macrophage- specific knockout mice. To complete this research, I will follow a detailed training plan laid out for me by my co- mentors and pursue my career goals. These include, among others, continuing my didactic training in areas relevant to my research, gaining experience in new laboratory techniques, developing effective written and oral communication skills, and building a professional network of scientists. Collectively, the training and research proposed here will not only fill a significant need for the development of ALD treatments but will also foster my development from a young scientist to a successful, independent investigator in the field of alcohol-induced organ disease.

项目摘要 酒精相关的肝病（ALD）造成肝病所有死亡的几乎一半。尽管 这种疾病的严重程度，对于任何疾病的任何阶段，尚无FDA批准的治疗方法， 强调阐明ALD进展的潜在机制以发展新的临界需求 疗法。有丝分裂原激活的蛋白激酶磷酸酶1（MKP-1）是双重型的原型成员 特异性磷酸酶在器官感染和损伤中具有关键作用，包括肝脏。较早的工作 表明改善的MKP-1水平与C-JUN N末端激酶（JNK）的激活有关 这会触发凋亡的级联反应，导致酒精引起肝损伤。但是，确切机制 MKP-1下调如何导致ALD中的肝细胞损伤和/或注射。而且， MKP-1在巨噬细胞中的作用和MAPK驱动的ALD中的作用从未被检查。我们的 初步研究表明，肝细胞中的MKP-1缺失增加了酒精诱导的肝损伤是NIAAA ALD的慢性加狂饮模型。数据还表明，肝细胞中的MKP-1删除会使它们感知到 TNFα诱导的毒性。作为一个有抱负的科学家，其目标是从事与酒精有关的多器官的职业 病理学，调查ALD发病机理中的新参与者不仅有助于满足这种医疗需求，还可以 作为一个出色的平台，可以从中学习如何成为一名成功的科学家。在帮助下 我的导师Gobejishvili博士（ALD基础研究专家）和McClain博士的监督（专家 该疾病的临床方面以及几个知识渊博的合作者，我将研究 ALD发病机理中MKP-1的酒精相关下调。这项研究的第一个目的是进一步 用肝细胞特异性MKP-1 knopockout小鼠在两只中表征MKP-1缺失在肝细胞中的作用 ALD的不同小鼠模型。我将从这些模型中分离出肝细胞和免疫电池，检查 MKP-1缺失对促炎性趋化因子产生和免疫细胞表型的影响并描述 肝细胞和免疫细胞健康和功能的变化。第二个目标将决定MKP-1的作用 特别是使用RAW 264.7巨噬细胞系，原代库普弗细胞和巨噬细胞 - 特定的淘汰小鼠。为了完成这项研究，我将遵循我的共同培训计划的详细培训计划 指导并追求我的职业目标。其中包括继续我在区域进行教学培训 与我的研究相关，获得新实验室技术的经验，开发有效的书面和口头 沟通技巧，并建立科学家的专业网络。共同培训和研究 这里提出的不仅会满足对ALD治疗的发展的重要需求，还将促进我的 从年轻科学家到酒精引起的成功独立研究者的发展 器官疾病。