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（由申请人提供）：Benita L. McVicker 于 1997 年获得内布拉斯加大学医学中心 (UNMC) 博士学位，并曾在 Drs. 的实验室工作。从那时起，Dean Tuma 和 Carol Casey 就在内布拉斯加州奥马哈退伍军人管理局医疗中心的肝脏研究室工作。肝脏研究小组在确定长期乙醇消耗可导致各种病理后果方面发挥了重要作用，但需要进一步阐明乙醇引起肝毒性病症的机制。最近，McVicker 博士及其同事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（无法用常规分离的肝细胞完成）的生成将能够评估酒精存在下与肝细胞极性和细胞凋亡改变相关的蛋白质运输事件（Fas/CD95死亡受体）。具体来说，McVicker 博士在这笔赠款中提出的研究目标包括 1) 进一步表征乙醇对 Fas 运输的影响，以及 2) 确定特定物质的作用 参与乙醇诱导的 Fas 易位的调节因子以及这些效应与 Fas 诱导的细胞凋亡机制的相关性。死亡受体运输和信号传导的研究以及 WIF-B 模型的使用是 McVicker 博士感兴趣的新领域。对于这些研究，她建议使用几种需要额外培训和专门指导的技术（即免疫组织化学和显微注射测定）。 McVicker 博士可以使用 UNMC 的共聚焦和流式细胞术核心实验室，并为这些方法的培训咨询提供支持。此外，她还获得华盛顿特区天主教大学 Pamela Tuma 博士（WIF-B 细胞使用专家）指导下学习特定技术的支持。总体而言，完成这项拟议工作将提供指导经验进行未来乙醇治疗后肝功能和生存的独立研究。