Regulation of IDDM by Proinflammatory and Th1-cytokines

促炎细胞因子和 Th1 细胞因子对 IDDM 的调节

• 批准号: 6984786
• 负责人: TORU MIYAZAKI
• 金额: $ 18.38万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6984786
• 关键词: NOD mouse; animal breeding; apoptosis; cell communication molecule; cell line; cytokine; dendritic cells; genetic library; genetically modified animals; helper T lymphocyte; inflammation; inhibitor /antagonist; insulin dependent diabetes mellitus; macrophage; pathologic process; toll like receptor

DESCRIPTION (provided by applicant): Insulin-dependent diabetes mellitus (IDDM) is characterized by the infiltration of T-lymphocytes into the islets of Langerhans of the pancreas (insulitis), followed by selective destruction of insulin-secreting beta cells leading to overt diabetes. Preliminarily, we observed the important association of IDDM with AIM factor (a secretion molecule we initially identified as an apoptosis inhibitory factor), which are: (1) AIM [-/-] mice backcrossed to non-obese diabetes (NOD) background showed complete prevention of IDDM; (2) AIM is expressed by infiltrating macrophages in the pancreatic islets from the very early stage of the disease; (3) AIM strongly induces TNF-alpha, IL-1 beta, IL-6 and IL-12 in macrophages and dendritic cells (DCs). Based on these, a hypothesis has emerged that AIM may accelerate IDDM by inducing pro-inflammatory- and type I- cytokines in initially infiltrating macrophages and DCs in the islets at the onset stage of the disease. The Specific Aim 1 and 2 are focused on establishing the propriety of the hypothesis by adding back the AIM expression in macrophages in AIM-null NOD mice expecting the disease recurrence (aim 1), and testing the impact of the cytokines downstream of AIM signaling on the disease acceleration by assessing whether the induction of the cytokines in macrophages and DCs via the Tet-inducible transgenic system may overcome the disease prevention in AIM-null NOD mice (aim 2). In addition, our recent result that AIM mediates Toll-signaling to induce the cytokines provoked an idea that the putative AIM-receptor may be a TolI/IL-1 receptor family member. In the Specific Aim 3, we will purify and characterize the AIM-receptor by expression screening of a cDNA library generated from macrophage cells. We also plan to create knockout mice of the AIM-receptor by disrupting the gene in the NOD-derived ES cells, as we generated AIM[-/-]/-NOD by using the cells. Our proposed studies will clarify the precise picture of the IDDM pathogenesis in the context of AIM, in particular, during the early stage of the disease, and thus will contribute to development of a new therapy via suppression of AIM. In addition, identification of AIM-receptor will shed light on the precise molecular machinery of AIM functions, as well as a new aspect of physiological function of the Toll-family.

描述（由申请人提供）：胰岛素依赖性糖尿病（IDDM）的特征是T淋巴细胞浸润到胰腺的朗格汉斯岛（胰岛炎），随后选择性破坏胰岛素分泌β细胞，导致明显的糖尿病。初步观察到IDDM与AIM因子（我们最初鉴定为凋亡抑制因子的分泌分子）之间存在重要关联，即：（1）与非肥胖糖尿病（NOD）背景回交的AIM[-/-]小鼠显示出完全预防IDDM； (2) AIM从疾病的早期阶段就由胰岛中浸润的巨噬细胞表达； (3) AIM强烈诱导巨噬细胞和树突状细胞(DC)中的TNF-α、IL-1β、IL-6和IL-12。基于这些，出现了一个假设，即在疾病发作阶段，AIM 可能通过在胰岛中最初浸润的巨噬细胞和 DC 中诱导促炎细胞因子和 I 型细胞因子来加速 IDDM。具体目标 1 和 2 的重点是通过在预期疾病复发的 AIM-null NOD 小鼠巨噬细胞中添加 AIM 表达来建立假设的正确性（目标 1），并通过评估通过 Tet 诱导的转基因系统在巨噬细胞和 DC 中诱导细胞因子是否可以克服疾病预防来测试 AIM 信号下游细胞因子对疾病加速的影响。 在 AIM 无效的 NOD 小鼠中（目标 2）。此外，我们最近的结果表明 AIM 介导 Toll 信号传导以诱导细胞因子，这引发了一个想法，即假定的 AIM 受体可能是 TolI/IL-1 受体家族成员。在具体目标 3 中，我们将通过对巨噬细胞生成的 cDNA 文库进行表达筛选来纯化和表征 AIM 受体。我们还计划通过破坏 NOD 衍生的 ES 细胞中的基因来创建 AIM 受体的敲除小鼠，因为我们使用这些细胞生成了 AIM[-/-]/-NOD。我们提出的研究将阐明 AIM 背景下 IDDM 发病机制的精确情况，特别是在疾病的早期阶段，从而有助于通过抑制 AIM 开发新疗法。此外，AIM受体的鉴定将揭示AIM功能的精确分子机制，以及Toll家族生理功能的新方面。