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状况，其是X综合征（代谢综合征，In-D）的基础。 胰岛素抵抗综合征）。X综合征与肥胖和2型糖尿病密切相关， 在美国，持续的肥胖症使这种异常的情况更加普遍。 本项目的一个主要主题是慢性炎症在hu的病因学中起着核心作用， 男性胰岛素抵抗状态。此外，我们提出巨噬细胞可能是这种炎症的启动者， 引起的胰岛素抵抗。支持这一概念的有力数据已经来自最近的研究 在巨噬细胞敲除的动物中，我们发现， 巨噬细胞导致小鼠整体胰岛素敏感性的状态，而过度激活（或去抑制） 巨噬细胞内的炎症途径导致葡萄糖耐受不良、高胰岛素血症和 胰岛素抵抗在这个提议中，我们计划进行一系列广泛的体外和体内研究， 旨在确定巨噬细胞在炎症诱导的胰岛素抵抗中的全部作用， derlying机制.我们将使用基因阵列的方法来阐明由PPARy调控的基因网络 以及sTs-Li脂肪细胞中的两种共阻遏物N-CoR和SMRT。我们已经开发了体外试验， 允许我们解剖出巨噬细胞行程的各个步骤，包括内皮细胞迁移， 趋化性和巨噬细胞与脂肪细胞的直接结合。此外，我们还建立了一个共同的文化， 在sTs-Li脂肪细胞中加入巨噬细胞引起细胞胰岛素抵抗的系统中， 提出了一系列的实验来确定这种效应的基本机制。最后，我们将大力 通过使用一些新的基因敲除模型和新的策略来进行我们的小鼠研究。在这个美联社- 我们将大量使用骨髓移植从一个给定的KO动物到辐射 C57 bl/6宿主以相对高的通量规模创建功能性巨噬细胞KO模型。我们还将 利用我们的lentisiRNA方法，用靶向特异性的lentisiRNA载体治疗正常骨髓， 巨噬细胞内的特异性炎症途径组分。这就造成了一个骨髓敲除， 然后这些细胞可以被移植并植入到受辐射的宿主小鼠中。这 进一步提高了我们方法的通量，并且还允许多重敲除的可能性， 评估组合效应。因此，这些结果应该确定相关的机制， 在胰岛素靶组织中引起整体胰岛素抵抗，也应该更好地阐明 目前可用的TZD的作用机制和作用的组织部位。