Identification of the adipogenic genes of Adenovirus 36

描述（由申请人提供）：肥胖在美国已经成为一个主要的健康问题。虽然许多肥胖的可能原因正在深入研究中，但感染因子的作用尚未得到很好的研究。人类36型腺病毒（Ad-36）在几种动物模型（小鼠、狨猴、鸡）中已被证明可导致体重过度增加和肥胖，并在人类血清流行病学研究中与肥胖有关。在细胞培养中，Ad-36还能加速3T3-L1前脂肪细胞向脂肪细胞的分化。本研究的目的是确定Ad-36基因（或基因）所需的这种效果。在Aim 1中，逆转录病毒载体将用于产生组成性表达各种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Aim 2中，将确定受脂肪生成Ad-36基因影响的调控途径。Western blot分析将用于确定蛋白水平和磷酸化状态。荧光显微镜将用于确定蛋白质的定位。在Aim 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.