rAAV-Mediated Metabolic Engineering in Vivo

rAAV 介导的体内代谢工程

• 批准号: 7190528
• 负责人: SERGEI ZOLOTUKHIN
• 金额: $ 24.83万
• 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/7190528
• 关键词: Adipocytes; Adipose tissue; Apoptosis; Body Weight; Breeding; Capsid; Ciliary Neurotrophic Factor; Complementary DNA; DNA Microarray Chip; Diabetes Mellitus; Diet; Ectopic Expression; Engineering; Fatty Liver; Fatty acid glycerol esters; Fibrosis; Gene Expression; Genes; Genetic; Histopathology; Hormones; Inflammatory; Injection of therapeutic agent; Injury; Insulin; Insulin Receptor; Insulin Resistance; Intervention; Intramuscular; Leptin; Link; Liver; Liver Cirrhosis; Longevity; Mediating; Messenger RNA; Metabolic; Metabolic Diseases; Modality; Modeling; Monitor; Morbid Obesity; Mus; Muscle; Mutation; Nonesterified Fatty Acids; Northern Blotting; Obesity; Organ; Pattern; Peripheral; Plasma; Portal vein structure; Rate; Rattus; Receptor Signaling; Recombinant adeno-associated virus (rAAV); Resistance; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Rodent Model; Route; Series; Serotyping; Signal Transduction; Signaling Pathway Gene; Sprague-Dawley Rats; Stress; Syndrome; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Transgenes; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Viral Vector; Weight; Weight Gain; Zucker Rats; adeno-associated viral vector; adiponectin; cytokine; diabetic; fibrogenesis; gene therapy; human TNF protein; improved; in vivo; indexing; inhibitor/antagonist; insulin sensitivity; insulin signaling; leukemia inhibitory factor; lipid metabolism; lipid transport; novel; promoter; response; vector; vector control

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）： 本项目的目的如下：（1）检测新型脂肪细胞激素脂联素通过重组腺相关病毒（rAAV）外周给药后，是否能够在具有特定遗传缺陷的肥胖啮齿类动物和饮食诱导肥胖（DIO）胰岛素抵抗大鼠中控制体重（BW）增加并纠正伴随的代谢紊乱。（2）确定脂联素的组成型表达是否增加肝脏的脂肪生成能力、增强胰岛素敏感性并诱导对脂肪萎缩性损伤和纤维形成的抗性;（3）证实脂联素的异位外周表达调节增强脂质转运、脂肪燃烧和消散的基因的表达，从而引起肌肉和肝脏脂质代谢的深刻变化。具体地，将在强组成型启动子控制下携带小鼠脂联素cDNA的一系列rAAV载体包装到三种不同AAV血清型的衣壳中：AAV 1、AAV 2和AAV 5。将在饮食诱导肥胖（DIO）Sprague-Dawley（SD）大鼠和Zucker糖尿病肥胖大鼠（fa/fa，ZDF）中检测这些载体。早先我们已经表明，编码脂肪抑制激素瘦素或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可以成功地用于在集中施用时抑制瘦SD大鼠的BW增加。在这里，我们建议在几种肥胖啮齿动物模型中使用外周途径的载体递送来评估脂联素的促氧化作用。主要读数将是间接监测血浆中的BW增量和胰岛素抵抗评估。为了鉴定脂联素异位表达调控的基因和信号通路，我们将使用从肝组织分离的mRNA的DNA微阵列芯片分析。这些研究的结果将不仅确定基因治疗是否是控制BW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找的联系。

项目成果

Metabolic hormones in saliva: origins and functions.

• DOI: 10.1111/odi.12015
• 发表时间: 2013-04
• 期刊: Oral diseases
• 影响因子: 3.8
• 作者: Zolotukhin S

Distribution of Y-receptors in murine lingual epithelia.

• DOI: 10.1371/journal.pone.0046358
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Hurtado MD;Acosta A;Riveros PP;Baum BJ;Ukhanov K;Brown AR;Dotson CD;Herzog H;Zolotukhin S
• 通讯作者: Zolotukhin S

Salivary PYY: a putative bypass to satiety.

• DOI: 10.1371/journal.pone.0026137
• 发表时间: 2011
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Acosta A;Hurtado MD;Gorbatyuk O;La Sala M;Duncan D;Aslanidi G;Campbell-Thompson M;Zhang L;Herzog H;Voutetakis A;Baum BJ;Zolotukhin S
• 通讯作者: Zolotukhin S

Salivary peptide tyrosine-tyrosine 3-36 modulates ingestive behavior without inducing taste aversion.

唾液肽酪氨酸-酪氨酸 3-36 调节摄取行为而不引起味觉厌恶。

• DOI: 10.1523/jneurosci.1064-13.2013
• 发表时间: 2013
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Hurtado,MariaD;Sergeyev,ValeriyG;Acosta,Andres;Spegele,Michael;LaSala,Michael;Waler,NickolasJ;Chiriboga-Hurtado,Juan;Currlin,SethW;Herzog,Herbert;Dotson,CedrickD;Gorbatyuk,OlegS;Zolotukhin,Sergei
• 通讯作者: Zolotukhin,Sergei