Application of Genetics and Physiological Genomics to Dissect Resistance to T1D

应用遗传学和生理基因组学剖析 T1D 抗性

• 批准号: 6990084
• 负责人: ANNE E. KWITEK
• 金额: $ 15.65万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6990084
• 关键词: diabetes mellitus genetics; functional /structural genomics; gene expression; gene expression profiling; genetic mapping; genetic susceptibility; genetically modified animals; insulin dependent diabetes mellitus; laboratory rat; lymphopenia; major histocompatibility complex; microarray technology

Type 1 diabetes (T1D) is one of the prototypical organ-specific autoimmune endocrinopathies that results in life-long dependency on daily insulin injection. Molecular genetic approaches have identified regions of the human, mouse, rat genomes that play a role in susceptibility to Type 1 Diabetes (T1D). While the major histocompatibility complex (MHC), the equivalent of the human leukocyte antigen (HLA), is the major locus responsible for susceptibility to T1D, there is a separate set of genes that contribute to susceptibility and end organ damage. We have been studying the genetic factors contributing to T1D in the BBDP rat, a model of spontaneous disease concurrent with lymphopenia. We have identified the location of five loci involved in T1D: Iddm2/lyp, lddm1/MHC, Iddm3, Iddm19, and a fifth locus on chromosome 4 (Iddm4?). Given the identification of the Ian5 mutation as the gene responsible for Iddm2/lyp, we now focus on the identification of the additional factors playing a role in T1D in the BB rat. Our mapping studies have identified four diabetogenic loci in addition to the MHC, which Projects 3 specifically addresses, in close collaboration with Project 1. The focus of Project 3 is to generate consomic and derived congenic strains for three additional factors, including Iddm3, Iddm19, and the susceptibility locus on chromosome 4 nearby Ian5 (Project 1) and to evaluate these new models for their role in T1D using gene expression profiling, comparative genomics, and sequence analysis. Furthermore, we are developing the technology to knock genes out in the rat for validation of positionally cloned genes. Specifically we will: 1. Continue positional cloning efforts for Iddm3 to identify sequence variants contributing to resistance to autoimmune diabetes. 2. Follow up a second resistance factor on chromosome 15. 3. Use novel 70-mer oligonucleotide and cDNA microarray platforms to evaluate genes and pathways affected by the diabetogenic factors identified in the BB rat. 4. Generate rat ENU-induced knockout strains to validate genes involved in autoimmune diabetes in the BB rat.

1 型糖尿病 (T1D) 是一种典型的器官特异性自身免疫性内分泌病，导致终生依赖每日胰岛素注射。分子遗传学方法已经确定了人类、小鼠和大鼠基因组中与 1 型糖尿病 (T1D) 易感性有关的区域。虽然主要组织相容性复合体 (MHC)（相当于人类白细胞抗原 (HLA)）是导致 T1D 易感性的主要基因座，但还有一组单独的基因导致易感性和终末器官损伤。我们一直在 BBDP 大鼠中研究导致 T1D 的遗传因素，BBDP 大鼠是一种伴有淋巴细胞减少的自发性疾病模型。我们已经确定了涉及的五个位点的位置 T1D：Iddm2/lyp、lddm1/MHC、Iddm3、Iddm19 和 4 号染色体上的第五个基因座（Iddm4？）。鉴于 Ian5 突变被鉴定为负责 Iddm2/lyp 的基因，我们现在重点鉴定在 BB 大鼠 T1D 中发挥作用的其他因素。我们的绘图研究已经确定了除 MHC 之外的四个糖尿病基因位点，项目 3 与项目 1 密切合作专门解决了这些基因座。项目 3 的重点是为另外三个因子生成 consomic 和衍生的同源菌株，包括 Iddm3、Iddm19、以及 Ian5 附近 4 号染色体上的易感位点（项目 1），并使用基因表达谱、比较基因组学和序列来评估这些新模型在 T1D 中的作用 分析。此外，我们正在开发敲除老鼠基因的技术 定位克隆基因的验证。具体来说，我们将： 1. 继续 Iddm3 的定位克隆工作，以确定有助于抵抗自身免疫糖尿病的序列变异。 2. 追踪 15 号染色体上的第二个耐药因子。 3. 使用新型 70 聚体寡核苷酸和 cDNA 微阵列平台来评估受 BB 大鼠中发现的糖尿病致病因素影响的基因和途径。 4. 生成大鼠 ENU 诱导的敲除菌株，以验证 BB 大鼠中与自身免疫性糖尿病有关的基因。