Functional Char. of Pro-Inflammatory Pathways Influencing Insulin Influencing Ins

功能特性

• 批准号: 7249790
• 负责人: jerrold Michael OLEFSKY
• 金额: $ 43.79万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7249790
• 关键词: Adhesions; Adipocytes; Adipose tissue; Affinity; Animals; Binding; Bone Marrow; Candidate Disease Gene; Cells; Chemotaxis; Chronic; Coculture Techniques; Collaborations; Condition; Data; Etiology; Gap Junctions; Gene Expression Profiling; Gene Targeting; Gene Transfer Techniques; Generations; Genes; Glucose Intolerance; Hyperinsulinism; Inflammation; Inflammatory; Insulin; Insulin Resistance; Knock-out; Knockout Mice; Ligands; Macrophage Activation; Mediating; Metabolic; Metabolic syndrome; Modeling; Molecular; Mus; Nexus (resin cement); Non-Insulin-Dependent Diabetes Mellitus; Nuclear Receptors; Numbers; Obesity; Pathogenesis; Pathway interactions; Phenotype; Play; Process; Production; Recruitment Activity; Repression; Resistance; Role; Series; Site; Specific qualifier value; Stem cells; Subfamily lentivirinae; Syndrome; System; Testing; Tissues; Transactivation; Transgenic Mice; Transplantation; United States; bone; cell type; combinatorial; concept; cytokine; desire; human NCOR1 protein; in vitro Assay; in vivo; insight; insulin sensitivity; knock-down; lentiviral-mediated; macrophage; man; migration; monocyte; mutant; novel; promoter; receptor; research study; rosiglitazone; small hairpin RNA

Insulin resistance is a common metabolic condition which underlies Syndrome X(Metabolic Syndrome, In- sulin Resistance Syndrome). Syndrome Xis strongly associated with obesity and Type 2 diabetes and the ongoing obesity epidemicin the United States makes this constellation ofabnormalities even more prevalent. An overarchingtheme in this project is that chronic inflammationplays a central role in the etiology of hu- man insulin resistant states. Further, wepropose that the macrophage canbe an initiator ofthis inflamma- tion-induced insulin resistance. Powerful data to support this concept has already come from recent studies of macrophageknockout animals in whichwe showed that disabling the inflammatorypathwaywithin macrophages leads to a state ofglobal insulin sensitivity in mice,whereas, hyperactivating (or derepressing) the inflammatory pathway within macrophages leads to a state ofglucose intolerance, hyperinsulinemia, and insulin resistance. In this proposal, we plan to conduct an extensive series ofin vitro and in vivo studies aimed at identifying the full role ofthe macrophagein inflammation-induced insulin resistance and the un- derlying mechanisms. We will use a gene array approachto elucidate the gene network regulated by PPARy and the two co-repressors N-CoRand SMRT in sTs-Li adipocytes. Wehave developed in vitro assays which allow us to dissect out the various steps ofthe macrophageitinerary, including endothelial transmigration, chemotaxis, and direct binding of macrophages to adipocytes. In addition, we have established a co-culture system in which addition ofmacrophages causes cellular insulin resistance in sTs-Li adipocytes, and a num- ber of experiments are proposed to identify the basic mechanisms ofthis effect. Finally, we willvigorously pursue our mouse studies by using a number ofnewknockout models as well as new strategies. In this ap- proach, we will make heavyuse ofbone marrowtransplantation from a given KOanimal into irradiated C57bl/6 hosts to create functional macrophage KO models on a relativelyhigh throughput scale. Wewill also utilize our lentisiRNAmethod oftreating normal bone marrow with lentisiRNAvectors targeted against spe- cific inflammatory pathway components within macrophages. This creates a bone marrowknockdownof the desired target, and these cells can then be transplanted and engraftedinto an irradiated host mouse. This further enhances the throughput of our approach and also allows the possibility of multiplexing knockouts to assess combinatorial effects. Assuch, these results should identify relevant mechanisms whereby activation of macrophages causes global insulin resistance in insulin target tissues and should also better elucidate the mechanisms of action, and tissue sites of action of currently available TZDs.

胰岛素抵抗是一种常见的代谢疾病，它是综合征X的基础（代谢综合征 苏林抵抗综合征）。综合征XIS与肥胖和2型糖尿病密切相关， 持续的肥胖症流行病使美国这一卑鄙的态度更加普遍。 该项目的总体列表是慢性炎症在HU-病因中起着核心作用 人类胰岛素抵抗状态。此外，巨噬细胞可以成为这种发炎的发起者 - TION诱导的胰岛素抵抗。支持这一概念的强大数据已经来自最近的研究 巨噬细胞销售动物，其中我们表明使炎症性行为无效 巨噬细胞导致小鼠全球胰岛素敏感性，而过度激活（或解压缩） 巨噬细胞内的炎症途径导致葡萄糖不耐症，高胰岛素血症和 胰岛素抵抗。在此提案中，我们计划进行大量的体外研究和体内研究 旨在确定巨噬细胞炎症引起的胰岛素抵抗的全部作用，并 衍生机制。我们将使用一种基因阵列方法来阐明由PPARY调节的基因网络 以及STS-LI脂肪细胞中的两个共抑制剂n-Corand SMRT。 Wehave开发了体外测定法 允许我们剖析巨噬细胞的各个步骤，包括内皮转移， 趋化性和巨噬细胞与脂肪细胞的直接结合。此外，我们已经建立了共同文化 在STS-LI脂肪细胞中导致细胞胰岛素耐药性的系统，并且数字 提出了一系列实验来确定这种效应的基本机制。最后，我们会进行审查 通过使用多种新销量模型以及新策略进行鼠标研究。在这个ap- proach，我们将从给定的koanimal中进行重复的骨髓术 C57BL/6主机以相对高的吞吐量量表创建功能性巨噬细胞KO模型。我们也将 利用我们的宽叶态小径治疗正常的骨髓，用针对Spe- 巨噬细胞中的可见炎症途径成分。这会产生骨髓敲击 所需的靶标，然后可以将这些细胞移植并植入辐照的宿主小鼠。这 进一步增强了我们方法的吞吐量，还可以使多路复用敲除可能 评估组合效应。 Assuch，这些结果应确定相关机制，以使激活 巨噬细胞在胰岛素靶组织中引起全球胰岛素抵抗，还应更好地阐明 作用机理和当前可用TZD的组织作用位点。