Identification of the adipogenic genes of Adenovirus 36

描述(申请人提供)：肥胖已成为美国的一个主要健康问题。虽然许多可能导致肥胖的原因正在深入调查中，但感染性因素的作用还没有得到很好的研究。人类36型腺病毒(Ad-36)已被证明在多种动物模型(小鼠、绒猴、鸡)中引起过度体重增加和肥胖，并在人类的血清流行病学研究中与肥胖有关。在细胞培养中，Ad-36还能促进3T3-L1前脂肪细胞向脂肪细胞分化。本研究的目的是确定这一效应所需的一个或多个Ad-36基因。在目标1中，逆转录病毒载体将被用于构建表达各种Ad-36开放阅读框的3T3-L1细胞系。这些Ad-36基因对脂肪细胞分化的影响将被确定。在目标2中，将确定成脂Ad-36基因影响的调控途径。蛋白质印迹分析将被用来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质的定位。在目的3中，将表达成脂基因Ad-36的慢病毒载体导入人前脂肪细胞，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.