sEH Inhibition in Alcoholic Liver Disease: A Novel Therapeutic Strategy

项目摘要 酒精性肝病（ALD）是最常见的肝脏疾病，占所有死亡人数的近一半 影响这个器官的疾病尽管如此，仍然没有FDA批准的治疗任何阶段的 疾病，强调需要发现新的治疗方法。作为一个有抱负的科学家， 在酒精相关的多器官病理学的职业生涯中，研究ALD治疗的新疗法不仅 帮助满足这一医疗需求，但也作为一个很好的平台，从中学习如何成为一个成功的 科学家在我的共同导师Kirpich博士（ALD基础研究专家）的帮助和监督下， 博士麦克莱恩（对疾病的临床方面的专家）以及几个知识渊博的合作者，我将 研究可溶性环氧化物水解酶（sEH）抑制作为治疗ALD的新治疗策略 使用临床前小鼠模型。这种方法已被应用于其他炎症性疾病，但没有 ALD，我们小组先前的工作显示的一种疾病，其特征是抗炎性 环氧化脂质（环氧化物），部分原因是它们被sEH快速降解。我已经为我们的公司 在ALD的慢性狂饮小鼠模型中口服sEH抑制剂的初步数据， 概括了人类ALD。通过本申请中提出的两个相关但独立的具体目标，我将 进一步评估sEH抑制的功效并探索其作用机制。目标1将使用药理学 和sEH的基因消除以进一步表征sEH抑制在ALD小鼠模型中的作用。目的2 将描述单个抗炎环氧化物对肝细胞变化的机制作用， 免疫细胞的健康和功能来解释体内观察到的变化。为了完成这项研究，我将遵循详细的 我会按照我的导师为我制定的培训计划，追求我为自己设定的职业目标。这些 包括，除其他外，继续我在与我的研究相关的领域的教学培训，获得经验， 新的实验室技术，发展有效的书面和口头沟通技巧，并建立一个 科学家的专业网络例如，为了评估肝损伤和细胞功能的变化，我将学习 阅读组织学肝载玻片，使用酶测定，建立ELISA，分离和培养原代肝细胞 和巨噬细胞，并学习流式细胞术。路易斯维尔大学的研究环境- 最先进的临床和转化研究大楼生产力很高，配备了专家教师， 研究人员愿意帮助我学习这些技术，并在我追求培训和职业生涯时为我提供建议 目标.我的学术部门，药理学与毒理学系，同样能够支持 我在课堂上的培训，并提供许多资源（如旅行基金，职业发展研讨会， #21453;的新课程，以确保我的成功。总的来说，这里提出的培训和研究不仅将填补一个空白， 这是对ALD治疗发展的重大需求，但也将促进我从一个年轻的发展， 科学家成为酒精引起的器官疾病领域成功的独立调查员。

Project Summary Alcoholic liver disease (ALD) is the most common liver disease and is responsible for nearly half of all deaths from diseases affecting this organ. Despite this, there remain no FDA-approved treatments for any stage of the disease, highlighting the need for the discovery of new therapies. As an aspiring scientist whose goal is to pursue a career in alcohol-related multi-organ pathology, investigating new therapies for ALD treatment 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 co-mentors Dr. Kirpich (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 soluble epoxide hydrolase (sEH) inhibition as a novel therapeutic strategy for the treatment of ALD using pre-clinical mouse models. This approach has been applied to other inflammatory diseases, but not to ALD, a disease shown by previous work in our group to be characterized by a loss of anti-inflammatory epoxygenated lipids (epoxides), in part due to their rapid degradation by sEH. I have generated substantial preliminary data with an orally-administered sEH inhibitor in a chronic-binge mouse model of ALD that recapitulates human ALD. Through two related, but independent, specific aims proposed in this application I will further assess the efficacy of sEH inhibition and explore its mechanisms of action. Aim 1 will use pharmacological and genetic ablation of sEH to further characterize the effects of sEH inhibition in mouse models of ALD. Aim 2 will describe the mechanistic effects of individual anti-inflammatory epoxides on changes in hepatocyte and immune cell health and function to explain changes seen in vivo. To complete this research, I will follow a detailed training plan laid out for me by my co-mentors and pursue the career goals that I have set for myself. 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. For example, to assess liver injury and changes in cellular function I will learn to read histological liver slides, use enzymatic assays, set up ELISAs, isolate and culture primary hepatocytes and macrophages, and learn flow cytometry. The research environment at the University of Louisville’s state-of- the-art Clinical and Translational Research Building is highly productive and staffed with expert faculty and research personnel willing to help me learn these techniques and advise me as I pursue my training and career goals. My academic department, the Department of Pharmacology & Toxicology, is equally positioned to support my training in the classroom and provide many resources (e.g. travel funds, career development seminars, and new courses) to ensure my success. 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.