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.

项目概要 酒精相关性肝病 (ALD) 导致近一半的肝病死亡。 由于这种疾病的严重程度，FDA 仍然没有批准针对该疾病任何阶段的治疗方法， 强调迫切需要阐明 ALD 进展的根本机制，以开发新的 丝裂原激活蛋白激酶磷酸酶 1 (MKP-1) 是双重疗法的典型成员。 特异性磷酸酶在器官炎症和损伤（包括肝脏）中发挥着关键作用。 显示 MKP-1 水平与 c-Jun N 末端激酶 (JNK) 的激活相关， 它会引发细胞凋亡级联反应，导致酒精引起的损伤，但其确切机制尚不清楚。 MKP-1 下调如何导致 ALD 中的肝细胞损伤和/或炎症是已知的。 MKP-1 在巨噬细胞和 ALD 中 MAPK 驱动的炎症中的作用尚未得到研究。 初步研究表明，肝细胞中 MKP-1 缺失会加重酒精引起的肝损伤，这是一种 NIAAA ALD 的慢性加暴饮暴食模型 数据还表明，肝细胞中 MKP-1 的缺失使它们变得敏感。 作为一名有抱负的科学家，他的目标是从事与酒精相关的多器官研究。 病理学方面，研究 ALD 发病机制中的新参与者不仅有助于满足这一医疗需求，而且 作为学习如何在帮助和帮助下成为一名成功科学家的绝佳平台。 我的导师 Gobejishvili 博士（ALD 基础研究专家）和 McClain 博士（ALD 基础研究专家）的监督 该疾病的临床方面）以及一些知识渊博的合作者，我将研究 酒精相关的 MKP-1 下调在 ALD 发病机制中的作用 本研究的首要目的是进一步研究。 使用肝细胞特异性 MKP-1 敲除小鼠在两个模型中表征 MKP-1 缺失在肝细胞中的作用 我将从这些模型中分离出肝细胞和免疫细胞，检查 MKP-1 缺失对促炎趋化因子产生和免疫细胞表型的影响并描述 第二个目标将确定 MKP-1 的作用。 特别是在巨噬细胞中，使用 RAW 264.7 巨噬细胞系、原代 Kupffer 细胞和巨噬细胞- 为了完成这项研究，我将遵循我的同事为我制定的详细训练计划。 导师并追求我的职业目标，其中包括继续我在各个领域的教学培训。 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，并建立一个专业的科学家网络。 这里提出的建议不仅将满足开发 ALD 治疗的重大需求，而且还将促进我的研究 从一名年轻科学家成长为酒精诱发领域成功的独立研究者 器官疾病。