rAAV-Mediated Metabolic Engineering in Vivo

rAAV 介导的体内代谢工程

• 批准号: 7023823
• 负责人: SERGEI ZOLOTUKHIN
• 金额: $ 25.57万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7023823
• 关键词: adeno associated virus group; anorexic agent; bioenergetics; collagen; complement; gene therapy; hormone regulation /control mechanism; hormone therapy; hormones; insulin sensitivity /resistance; laboratory rat; lipid metabolism; metabolism; microarray technology; nonhuman therapy evaluation; nutrition related tag; obesity; pathogenic diet; recombinant virus; transfection /expression vector; weight control; weight gain

DESCRIPTION (provided by applicant): The proposed project has the following objectives: (1) to test whether the novel adipocyte hormone adiponectin, when administered peripherally via recombinant adeno-associated virus (rAAV), can control body weight (BW) gain and correct the attendant metabolic disorders for extended periods of time in obese rodents with specific genetic defects and in diet-induced obese (DIO) insulin-resistant rats; (2) to determine whether constitutive expression of adiponectin increases lipogenic capacity of the liver, augments insulin sensitivity, and induces resistance to lipoatrophic injury and fibrogenesis; (3) to confirm that ectopic peripheral expression of adiponectin modulates expression of genes that enhance lipid transport, fat combustion and dissipation, thereby inducing profound changes in muscle and liver lipid metabolism. Specifically, a series of rAAV vectors carrying mouse adiponectin cDNA under the control of a strong constitutive promoter will be packaged into capsids of three different AAV serotypes: AAV1, AAV2 and AAV5. These vectors will be tested in diet-induced obese (DIO) Sprague-Dawley (SD) rats and Zucker diabetic fatty rats (fa/fa, ZDF). Earlier we have shown that a rAAV encoding either the lipostatic hormone leptin, or the pleiotropic cytokines ciliary neurotrophic factor (CNTF), or leukemia inhibitory factor (LIF) could be successfully used to curb the BW gain in lean SD rats when administered centrally. Here we propose to evaluate an anorexigenic effect of adiponectin using peripheral routes of vector delivery in several obese rodent models. The primary readouts will be assessments of BW gain and insulin resistance monitored indirectly in plasma. In order to identify genes and signaling pathways modulated in response to adiponectin ectopic expression we will use DNA microarray chip analysis of mRNA isolated from liver tissue. The results of these studies will not only determine if gene therapy is a viable long-term therapeutic strategy to control BW and insulin resistance, but may also establish that rAAV-delivered adiponectin is an effective therapeutic modality in treating obesity and diabetes caused by environmental (DIO) or genetic factors. Results of these studies should also elucidate the function of a novel white fat-derived hormone, adiponectin, as the long-sought link between obesity and insulin resistance.

描述(由申请人提供)： 该项目有以下目标：(1)测试新型脂肪细胞激素脂联素通过重组腺相关病毒(RAAV)外周给药是否能够控制具有特定遗传缺陷的肥胖大鼠和饮食诱导的肥胖(DIO)胰岛素抵抗大鼠长时间的体重增加和纠正伴随的代谢紊乱；(2)确定脂联素的结构性表达是否增加肝脏的成脂能力，增强胰岛素敏感性，并诱导对脂肪萎缩损伤和纤维化的抵抗；(3)证实脂联素的异位外周表达调控促进脂肪转运、脂肪燃烧和消散的基因表达，从而引起肌肉和肝脏脂肪代谢的深刻变化。具体地说，一系列携带小鼠脂联素基因的重组腺相关病毒载体将被包装成三种不同AAV型的衣壳：AAV1、AAV2和AAV5。这些载体将在饮食诱导肥胖(DIO)Spraogue-Dawley(SD)大鼠和Zucker糖尿病肥胖大鼠(FA/FA，ZDF)中进行测试。早些时候，我们已经证明，编码脂肪激素瘦素、多效性细胞因子睫状神经营养因子(CNTF)或白血病抑制因子(LIF)的rAAV在中枢给药时可以成功地抑制瘦SD大鼠的体重增加。在这里，我们建议评估脂联素在几种肥胖啮齿动物模型中使用载体传递的外围途径的厌食效应。主要读数将是间接监测血浆中体重增加和胰岛素抵抗的评估。为了找出脂联素异位表达调控的基因和信号通路，我们将利用DNA微阵列芯片分析从肝组织中分离出的mRNA。这些研究的结果不仅将确定基因疗法是否是控制体重和胰岛素抵抗的可行的长期治疗策略，而且还可能确定rAAV递送的脂联素是治疗由环境(DIO)或遗传因素引起的肥胖和糖尿病的有效治疗方式。这些研究的结果也应该阐明一种新的白色脂肪衍生激素--脂联素的功能，它是肥胖和胰岛素抵抗之间长期寻求的联系。