BLR&D Research Career Scientist Award

BLR

• 批准号: 10047244
• 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/10047244
• 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; Fatty Liver; 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; 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酒精暴露如何导致高尔基体功能受损 仪器。高尔基体(也称为高尔基体或高尔基复合体)将蛋白质包裹到膜上。 在囊泡被送到它们的目的地之前，在细胞内结合囊泡。因此，这个细胞器驻留在 分泌途径、溶酶体途径和内吞途径的交叉点；众所周知它具有特殊的重要性 在处理蛋白质以供分泌时。我们实验室以前的工作已经发现了多个缺陷 酒精摄入后的内吞、蛋白质运输和分泌，但我们到目前为止还没有检查 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高 在退伍军人和全国退伍军人医疗中心超过一半的医疗入院人数中 与酗酒有关，我们正在集中努力确定干预的潜在目标 在长期饮酒后发生的进行性损伤期间，也许高尔基人会证明 成为这样的靶子。对我们的研究来说，最重要的是一个小的GTP酶Rab3D的作用，它参与了 在胞吐、分泌和囊泡运输中。我们已经证明了Rab3D的蛋白质含量显著 在酒精注射后减少，最近我们获得了令人兴奋的新的初步数据，乙醇- Rab3D功能受损在高尔基体的解体和碎裂中起重要作用。这些研究 在这项申请中提出的建议将扩展我们正在进行的乙醇如何改变肝细胞生物学的研究， 特别是在蛋白质加工方面，以检查其在通过高尔基体运输中的作用。我们为您提供一个 酒精诱导的高尔基体形态重塑是高尔基体后损伤的概念 贩运，这导致利用跨高尔基膜形成自噬小体。为了这个 工作中，我们提出了以下假设：酒精暴露通过其 碎裂和自噬介导的高尔基体膜裂解，导致受损的内吞和 胞外蛋白运输。小分子GTP酶Rab3D的分布和功能改变起着至关重要的作用 在这些变化中扮演的角色。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 研究乙醇给药前后Rab3D在体内和体外肝细胞中的分布。 为AIM 2提出的研究将确定Rab3D在生理相关的肝脏运输中的作用 蛋白质。在这些研究之后，将为目标3建议进行实验，在那里我们将确定Etoh- 诱导高尔基体解体和碎片化有助于自噬小体的形成及其如何改变Rab3D 功能影响肝细胞自噬。总之，成功完成这些目标将是 乙醇对高尔基体解体的影响，以及改变的Rab3D在这一过程中的作用。我们会的 能够将酒精介导的肝细胞转运受损的机制与高尔基体功能受损联系起来 并提供可能导致旨在减少或消除肝脏损伤的治疗策略的关键信息。 在这项工作中，与我一起工作的还有三位杰出的合作调查员(彼得罗相博士、托姆斯博士和拉西尼尼博士)，后者 其中两人是奥马哈退伍军人管理局的年轻调查人员。我还与多个退伍军人管理局进行了协作 研究人员，正如我的研究计划中概述的那样，这些合作一直很有成效。这个 酒精如何损害高尔基体和脂滴等重要细胞器的功能 为酒精性和非酒精性脂肪的治疗干预提供新的途径 肝病。我们的贡献意义重大，因为这是为 脂肪肝治疗方法的发展。此外，我们的发现还将对以下方面产生更广泛的影响 以肝损伤为特征的其他肝病。凭借我的专业知识和协作 我们预计，我们将在这些研究中取得成功，并能够为改进 通过确定酒精性肝损伤的相关机制，为退伍军人患者提供医疗保健。