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.

我目前的工作的目的是检查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体配合物）将蛋白质包装到膜上 在将囊泡发送到其目的地之前，在细胞内的绑定囊泡。因此，这个细胞器驻留在 分泌，溶酶体和内吞途径的交集；众所周知 在处理蛋白质以进行分泌。我们实验室的先前工作已经确定了多个缺陷 饮酒后的内吞作用，蛋白质运输和分泌，但直到现在我们还没有检查 在这些过程中改变高尔基体功能的作用。因为酒精性肝病的发生率更大 在退伍军人人口和全国弗吉尼亚州医疗中心的所有医疗入院中，有一半以上 与滥用酒精有关，我们正在集中精力识别潜在目标以干预 在慢性酒精给药后发生的进行性伤害期间，也许高尔基 成为这样的目标。对我们的研究至关重要的是小gtpase rab3d的作用，它涉及 在胞吐，分泌和囊泡运输中。我们已经表明Rab3d蛋白含量显着 饮酒后减少，最近我们获得了令人兴奋的新初步数据 受损的RAB3D功能在高尔基的混乱和碎片化中起着重要作用。研究 在此应用中提出的将扩展我们对乙醇如何改变肝细胞生物学的持续调查， 特别是在蛋白质加工方面，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形态重塑如何是高尔基的重大损害的概念 贩运，这导致了跨加利基膜形成自噬体的利用。为了这 工作，我们提出以下假设：乙醇暴露会导致高尔基的混乱 碎裂和自噬介导的高尔基膜裂解，导致内吞和 外生蛋白运输。小gtpase rab3d的分布和功能改变了关键 在这些改变中的作用。为了审查我们的假设，我们提出了三个具体目标。在目标1中，我们将 表征在ETOH给药之前和之后，体外和体内Rab3d的分布。 AIM 2提出的研究将在生理相关的肝脏运输中确立RAB3D的作用 蛋白质。这些研究将在AIM 3提出的实验之后进行，我们将确定是否eTOH- 诱发的高尔基混乱和碎片化有助于自噬体形成以及Rab3d的改变如何 功能会影响肝细胞自噬。总之，这些目标的成功完成将表征 ETOH对高尔基混合的影响，并在此过程中建立了改变Rab3d的作用。我们会的 能够将酒精介导的肝细胞运输障碍的机制与高尔基的功能相关联 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，我加入了三位出色的共同研究员（Petrosyan博士，Thomes和Rasineni），后者 两名是位于奥马哈弗吉尼亚州的年轻调查员。我也与多个VA合作 正如我的研究计划中概述的那样，调查人员的生产力很高。这 探索酒精如何损害重要细胞器的功能，例如高尔基和脂质液滴 为开发酒精和非酒精性脂肪的治疗干预措施提供新的途径 肝病。我们的贡献很重要，因为这是为提供翻译知识的关键步骤 开发脂肪肝病的疗法。此外，我们的发现还将对 其他以肝损伤为特征的肝病。有了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，并能够为改进做出贡献 通过鉴定酒精肝损伤的机制，为退伍军人患者提供医疗保健。