Alcohol & FAS-Mediated Apoptosis in Polarized Liver Cell

描述（由申请人提供）：Benita L. McVicker于1997年获得内布拉斯加州大学医学中心（UNMC）的博士学位，曾在博士实验室工作。迪安·图马和卡罗尔·凯西在位于内布拉斯加州奥马哈的退伍军人医疗中心的肝脏研究部门工作。肝脏研究小组在确定慢性乙醇摄入可导致多种病理后果方面发挥了重要作用，但需要进一步阐明乙醇导致肝毒性疾病的机制。最近，McVicker博士及其同事已经确定使用成纤维细胞/肝癌杂交细胞系（wi - b）作为研究乙醇对细胞过程影响的模型。产生这样一个生理上重要的极化模型（不能用常规分离的肝细胞完成）将能够评估酒精存在下的蛋白质运输事件（Fas/CD95死亡受体）与肝细胞极性改变和凋亡的关系。具体来说，McVicker博士在这项资助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；2)确定特异性的作用

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.