BLR&D Research Career Scientist Award

BLR

• 批准号: 10293582
• 负责人: Carol A. Casey
• 金额: --
• 依托单位: OMAHA VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-10-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10293582
• 关键词: Address; Admission activity; Affect; Alcohol abuse; Alcoholic Fatty Liver; Alcoholic Liver Diseases; Alcohols; American; Animal Model; Apoptotic; Area; Asialoglycoprotein Receptor; Autophagocytosis; Autophagosome; Award; Basic Science; Biochemical; Biology; Book Chapters; Carcinoembryonic Antigen; Cells; Cellular biology; Cessation of life; Chronic; Clinical; Collaborations; Country; Cytolysis; Data; Defect; Destinations; Development; Disease Progression; Endocytosis; Ensure; Ethanol; Ethanol Metabolism; Exocytosis; Family; Fibronectins; Functional disorder; General Population; Goals; Golgi Apparatus; Grant Review; Guanosine Triphosphate Phosphohydrolases; Healthcare; Heavy Drinking; Hepatic; Hepatocyte; Hepatotoxicity; Impairment; In Vitro; Incidence; Inflammatory Response; Injury; Investigation; Journals; Knowledge; Laboratories; Lead; Ligands; Link; Lipids; Liver; Liver Carbohydrate-Binding Protein; Liver diseases; Manuscripts; Mediating; Medical; Medical center; Membrane; Mentors; Military Personnel; Mission; Monitor; Monomeric GTP-Binding Proteins; Morphology; National Institute on Alcohol Abuse and Alcoholism; Organelles; Pathogenesis; Pathway interactions; Patients; Peer Review; Physiological; Play; Process; Program Development; Protein Secretion; Proteins; Publishing; Recycling; Research; Research Personnel; Role; Scientist; Seminal; Signal Transduction; Societies; Therapeutic; Therapeutic Intervention; Time; Toxic effect; Toxin; Training; United States National Institutes of Health; Vesicle; Veterans; Work; World Health; alcohol consequences; alcohol effect; alcohol exposure; base; career; chronic liver disease; design; experimental study; fatty liver disease; improved; in vivo; insight; liver injury; liver transplantation; member; military veteran; non-alcoholic; non-alcoholic fatty liver disease; novel; population health; problem drinker; programs; protein transport; receptor; receptor mediated endocytosis; receptor recycling; special interest group; symposium; therapeutic development; therapeutic target; therapeutically effective; therapy development; trafficking; trans-Golgi Network; undergraduate student; uptake

The goal of my current work is to examine how ethanol exposure results in impaired function of the Golgi apparatus. The Golgi apparatus (also called the Golgi body or Golgi complex) packages proteins into membrane bound vesicles inside the cell before the vesicles are sent to their destination. As such, this organelle resides at the intersection of the secretory, lysosomal, and endocytic pathways; it is known to be of particular importance in processing proteins for secretion. Previous work from our laboratory has identified multiple defects in endocytosis, protein trafficking, and secretion, after alcohol administration, but we have not until now, examined a role for altered Golgi function in these processes. Because the incidence of alcoholic liver disease is greater in the Veteran population and more than half of all medical admissions in VA Medical Centers across the country are linked to alcohol abuse, we are focusing efforts towards the identification of potential targets to intervene during the progressive injury which occurs after chronic alcohol administration, and perhaps the Golgi will prove to be such a target. Of central importance to our study is the role of a small GTPase, Rab3D, which is involved in exocytosis, secretion and vesicle trafficking. We have shown that Rab3D protein content was significantly decreased after alcohol administration, and recently we have obtained exciting new preliminary data that ethanol- impaired Rab3D function plays an important role in Golgi disorganization and fragmentation. The studies proposed in this application will extend our ongoing investigation of how ethanol alters hepatocyte biology, specifically in protein processing, to an examination of its role in transport through the Golgi. We provide a concept as to how alcohol-induced remodeling of Golgi morphology is a significant impairment of post-Golgi trafficking, and this leads to utilization of trans-Golgi membranes for the formation of autophagosomes. For this work, we present the following hypothesis: Ethanol exposure contributes to Golgi disorganization via its fragmentation and autophagy-mediated Golgi membrane lysis, leading to impaired endocytic and exocytic protein trafficking. Altered distribution and function of the small GTPase Rab3D plays a critical role in these alterations. To examine our hypothesis, we have proposed three specific aims; in Aim 1 we will characterize the distribution of Rab3D in vitro and in vivo in liver cells before and after EtOH administration. Studies proposed for Aim 2 will establish a role for Rab3D in the transport of physiologically relevant hepatic proteins. These studies will be followed by experiments proposed for Aim 3 where we will determine if EtOH- induced Golgi disorganization and fragmentation contribute to autophagosome formation and how altered Rab3D function affects hepatocyte autophagy. Altogether, successful completion of these aims will characterize the effect of EtOH on Golgi disorganization, and establish a role for altered Rab3D during this process. We will be able to correlate mechanisms of alcohol-mediated liver cell trafficking impairments with impaired Golgi function and provide key information that could lead to therapeutic strategies aimed at reducing or eliminating liver injury. In this work, I am joined by three outstanding co-investigators (Drs. Petrosyan, Thomes and Rasineni), the latter two are young investigators based at the Omaha VA. I also have collaborative effort with multiple VA investigators, as outlined in my research plan, and these collaborations have been highly productive. The exploration of how alcohol impairs function of important organelles such as the Golgi and lipid droplets will provide new avenues for the development of therapeutic interventions for both alcoholic and non-alcoholic fatty liver disease. Our contribution is significant since this is a critical step to provide translational knowledge for the development of therapies for fatty liver disease. Additionally, our findings will also have broader implications for other hepatic diseases characterized by hepatic injury. With the expertise and collaborations available to my group, we anticipate that we will be successful in these studies and will be able to contribute to improved healthcare for Veteran patients by the identification of mechanisms involved in the alcoholic liver injury.

我目前工作的目标是研究乙醇暴露如何导致高尔基体功能受损 设备。高尔基体（也称为高尔基体或高尔基复合体）将蛋白质包装到膜中 在囊泡被发送到目的地之前，将囊泡结合在细胞内。因此，该细胞器位于 分泌、溶酶体和内吞途径的交叉点；众所周知，它特别重要 处理蛋白质以供分泌。我们实验室之前的工作已经发现了多个缺陷 饮酒后的内吞作用、蛋白质运输和分泌，但我们直到现在才检查 高尔基体功能改变在这些过程中的作用。因为酒精性肝病的发病率更大 退伍军人人口以及全国退伍军人事务部医疗中心一半以上的就诊人数 与酗酒有关，我们正在重点努力确定潜在的干预目标 在长期饮酒后发生的进行性损伤期间，也许高尔基体会证明 成为这样的目标。对我们的研究来说最重要的是一种小型 GTPase Rab3D 的作用，它参与 胞吐作用、分泌和囊泡运输。我们已经证明 Rab3D 蛋白含量显着 饮酒后下降，最近我们获得了令人兴奋的新初步数据，即乙醇- Rab3D 功能受损在高尔基体解体和破碎中起着重要作用。研究 本申请中提出的建议将扩展我们正在进行的关于乙醇如何改变肝细胞生物学的研究， 特别是在蛋白质加工中，检查其在通过高尔基体运输中的作用。我们提供一个 关于酒精诱导的高尔基体形态重塑如何对后高尔基体造成显着损害的概念 贩运，这导致利用跨高尔基体膜来形成自噬体。为了这 工作中，我们提出以下假设：乙醇暴露通过其 碎片和自噬介导的高尔基体膜裂解，导致内吞和 胞吐蛋白运输。小 GTPase Rab3D 的分布和功能的改变起着至关重要的作用 在这些改变中发挥的作用。为了检验我们的假设，我们提出了三个具体目标；在目标 1 中，我们将 表征 EtOH 给药前后 Rab3D 在体外和体内肝细胞中的分布。 为目标 2 提出的研究将确定 Rab3D 在生理相关肝脏转运中的作用 蛋白质。这些研究之后将进行针对目标 3 提出的实验，我们将确定 EtOH- 诱导的高尔基体解构和断裂有助于自噬体形成以及 Rab3D 的改变 功能影响肝细胞自噬。总而言之，成功完成这些目标将成为 EtOH 对高尔基体解体的影响，并确定在此过程中改变的 Rab3D 的作用。我们将会 能够将酒精介导的肝细胞运输损伤的机制与受损的高尔基体功能相关联 并提供可能导致旨在减少或消除肝损伤的治疗策略的关键信息。 在这项工作中，我与三位杰出的联合研究员（Petrosyan、Thomes 和 Rasineni 博士）一起参与了工作，后者 其中两人是奥马哈退伍军人管理局的年轻调查员。我还与多个 VA 进行了合作 正如我的研究计划中所述，这些合作非常富有成效。这 探索酒精如何损害高尔基体和脂滴等重要细胞器的功能 为开发酒精性和非酒精性脂肪的治疗干预措施提供新途径 肝脏疾病。我们的贡献是重大的，因为这是为以下领域提供翻译知识的关键一步： 脂肪肝疾病治疗方法的开发。此外，我们的研究结果还将对以下领域产生更广泛的影响： 以肝损伤为特征的其他肝脏疾病。凭借我的专业知识和合作 小组，我们预计我们将在这些研究中取得成功，并将能够为改善 通过识别酒精性肝损伤的机制来为退伍军人患者提供医疗保健。