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 酒精相关性肝病（ALD）是近一半的肝病死亡原因。尽管 这种疾病的严重程度，仍然没有FDA批准的治疗方法用于疾病的任何阶段， 强调迫切需要阐明ALD进展的潜在机制，以开发新的 治疗丝裂原活化蛋白激酶磷酸酶1（MKP-1）是双磷酸酶的原型成员。 特异性磷酸酶在器官炎症和损伤中起关键作用，包括肝脏。早期工作 显示MKP-1水平的降低与c-Jun N-末端激酶（JNK）的激活有关， 其触发导致酒精诱导的肝损伤的凋亡级联。然而， 在ALD中MKP-1下调如何导致肝细胞损伤和/或炎症是已知的。而且 MKP-1在巨噬细胞中的作用和MAPK驱动的炎症在ALD中的作用从未被研究过。我们 初步研究表明，肝细胞中MKP-1的缺失增加了酒精诱导的肝损伤， 慢性酒精中毒加酗酒模型。数据还表明，肝细胞中的MKP-1缺失使其对以下物质敏感： TNFα诱导的毒性。作为一个有抱负的科学家，他的目标是从事与酒精有关的多器官 研究ALD发病机制的新参与者不仅有助于满足这种医学需求， 作为一个很好的平台，从中学习如何成为一个成功的科学家。的帮助和 在我的导师Gobejishvili博士（酒精性肝脏病基础研究专家）和McClain博士（酒精性肝脏病基础研究专家）的监督下 疾病的临床方面）以及几个知识渊博的合作者，我将调查的作用， 酒精相关的MKP-1在ALD发病机制中的下调。本研究的第一个目的是进一步 在两个实验中，使用肝细胞特异性MKP-1敲除小鼠来表征MKP-1缺失在肝细胞中的作用。 不同的ALD小鼠模型。我将从这些模型中分离肝细胞和免疫细胞， MKP-1缺失对促炎趋化因子产生和免疫细胞表型影响 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 特别是在使用RAW 264.7巨噬细胞系、原代库普弗细胞和巨噬细胞的巨噬细胞中- 特异性基因敲除小鼠为了完成这项研究，我将遵循我的合作伙伴为我制定的详细培训计划。 导师和追求我的职业目标。这些措施包括，除其他外，继续我的教学培训领域 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。总的来说， 这里提出的不仅将填补ALD治疗发展的重大需求，而且还将促进我的研究。 从一个年轻的科学家发展到一个成功的，独立的研究人员在酒精诱导的领域， 器官疾病