Alcohol & FAS-Mediated Apoptosis in Polarized Liver Cell

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• 批准号: 7045785
• 负责人: BENITA L. MCVICKER
• 金额: $ 10.66万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7045785
• 关键词: CD95 molecule; JUN kinase; apoptosis; biological signal transduction; cell line; cellular polarity; chemosensitizing agent; confocal scanning microscopy; cytotoxicity; enzyme activity; ethanol; female; flow cytometry; hepatotoxin; immunocytochemistry; intracellular transport; laboratory mouse; liver cells; microinjections; phosphorylation; posttranslational modifications; protein kinase C; protein structure function; protein transport; tissue /cell culture; toxicology; zinc

DESCRIPTION (provided by applicant): Benita L. McVicker received her PhD from The University of Nebraska Medical Center (UNMC) in 1997 and has worked in the laboratories of Drs. Dean Tuma and Carol Casey in the Liver Study Unit at the VA Medical Center in Omaha, Nebraska since that time. The Liver Study Unit has been instrumental in establishing that chronic ethanol consumption can lead to a variety of pathological consequences, yet further clarification of the mechanism(s) by which ethanol causes hepatotoxic conditions is required. Recently, Dr. McVicker and co-workers have identified the use of a fibroblast/hepatoma hybrid cell line (WIF-B) as a model for studying the effects of ethanol on cellular processes. The generation of such a physiologically important polarized model (that cannot be accomplished with regular isolated hepatocytes) will enable the evaluation of protein trafficking events (Fas/CD95 death receptor) in the presence of alcohol in relation to alterations in hepatocyte polarity and apoptosis. Specifically, the goals of Dr. McVicker's research presented in this grant include 1) further characterization of ethanol's effects on Fas trafficking and 2) to identify the role of specific regulators that are involved in ethanol-induced Fas translocation and the correlation of these effects with Fas-induced apoptosis mechanisms. The study of death receptor trafficking and signaling as well as the use of the WIF-B model are new areas of interest to Dr. McVicker. For these studies, she has proposed to use several techniques (i.e. immunohistochemistry and microinjection assays) that will require additional training and specialized instruction. Dr. McVicker has access to the confocal and flow cytometry core labs at UNMC and has support for training consultations in those methods. In addition, she has support to learn specific techniques under the direction of Dr. Pamela Tuma (an expert in the use of WIF-B cells) at The Catholic University in Washington D.C. Overall, the completion of this proposed work will provide the mentored experience to progress to future independent study of liver function and survival following ethanol treatment.

描述(申请人提供)：Benita L.McVicker于1997年在内布拉斯加州大学医学中心(UNMC)获得博士学位，此后一直在内布拉斯加州奥马哈市退伍军人医学中心肝脏研究室的Dean Tuma博士和Carol Casey博士的实验室工作。肝脏研究小组在确定长期饮酒可导致多种病理后果方面发挥了重要作用，但仍需进一步澄清酒精导致肝毒性的机制(S)。最近，麦克维克博士和他的同事已经确定使用成纤维细胞/肝癌杂交细胞系(WIF-B)作为研究乙醇对细胞过程影响的模型。这种生理上重要的极化模型的建立(这不能用常规的分离肝细胞来完成)将能够评估在酒精存在下与肝细胞极性和凋亡变化相关的蛋白质转运事件(Fas/CD95死亡受体)。具体地说，麦克维克博士在这项研究中提出的目标包括1)进一步表征乙醇对Fas运输的影响，以及2)确定特定的 参与乙醇诱导Fas移位的调控因子及其与Fas诱导的细胞凋亡机制的相关性。对死亡受体交易和信号的研究以及WIF-B模型的使用是麦克维克博士感兴趣的新领域。对于这些研究，她建议使用几种技术(即免疫组织化学和微量注射分析)，这将需要额外的培训和专门指导。McVicker博士可以使用UNMC的共焦和流式细胞仪核心实验室，并为这些方法的培训咨询提供支持。此外，她还得到了在华盛顿天主教大学帕梅拉·图马博士(WIF-B细胞使用方面的专家)的指导下学习具体技术的支持。总的来说，这项拟议工作的完成将为未来乙醇治疗后肝功能和存活率的独立研究提供指导经验。