Identification of the adipogenic genes of Adenovirus 36

描述（由申请人提供）：肥胖已成为美国的主要健康问题。虽然肥胖的许多可能原因正在深入调查中，但感染因子的作用尚未得到充分研究。人腺病毒36型（Ad-36）已显示在几种动物模型（小鼠、绒猴、鸡）中引起过度体重增加和肥胖，并且在人的血清流行病学研究中与肥胖有关。Ad-36还在细胞培养中加速3 T3-L1前脂肪细胞向脂肪细胞的分化。本研究的目的是鉴定这种效应所需的Ad-36基因（或基因）。在目的1中，逆转录病毒载体将用于产生组成型表达各种Ad-36开放阅读框的3 T3-L1细胞系。将确定这些Ad-36基因对脂肪细胞分化的影响。在目标2中，将鉴定受成脂Ad-36基因影响的调控途径。蛋白质印迹分析将用于确定蛋白质水平和磷酸化状态。荧光显微镜将用于确定蛋白定位。在目标3中，将用表达成脂Ad-36基因的慢病毒载体转导人前脂肪细胞，以确定其对人前脂肪细胞分化的影响。

DESCRIPTION (provided by applicant): Obesity has become a major health problem in the United States. Although many of the possible causes of obesity are under intensive investigation, the role of infectious agents has been not been well studied. Human adenovirus type 36 (Ad-36) has been shown to cause excessive weight gain and adiposity in several animal models (mice, marmosets, chickens) and is linked to obesity in seroepidemiological studies of humans. Ad-36 also accelerates the differentiation of 3T3-L1 preadipocytes to adipocytes in cell culture. The purpose of this study is to identify the Ad-36 gene (or genes) required for this effect. In Aim 1, retroviral vectors will be used to produce 3T3-L1 cell lines constitutively expressing various Ad-36 open reading frames. The effect of these Ad-36 genes on adipocytic differentiation will be determined. In Aim 2, the regulatory pathways affected by the adipogenic Ad-36 gene will be identified. Western blot analysis will be used to determine protein levels and phosphorylation state. Fluorescence microscopy will be used to determine protein localization. In Aim 3, human preadipocytes will be transduced with lentiviral vectors expressing the adipogenic Ad-36 gene to determine its effects on human preadipocyte differentiation.