A new genetic approach to identify the Idd9.3 type 1 diabetes susceptibility gene

鉴定 Idd9.3 1 型糖尿病易感基因的新遗传学方法

• 批准号: 9303282
• 负责人: Yi-Guang Chen
• 金额: $ 19万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-23 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9303282
• 关键词: Affect; Alleles; Animal Model; Autoimmune Diseases; Autoimmune Process; Backcrossings; Biological Models; Breeding; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Candidate Disease Gene; Chromosome Mapping; Chromosomes, Human, Pair 4; Code; Complex; Congenic Mice; Congenic Strain; Development; Diabetes Mellitus; Disease; Distal; FOXP3 gene; Frequencies; Generations; Genes; Genetic; Genetic Determinism; Genetic Engineering; Genetic Polymorphism; Genetic study; Genome; Germ Lines; Goals; Human; Human Genetics; Human Genome; Hybrids; Immune; Immune Tolerance; Immunosuppressive Agents; Inbred NOD Mice; Inbred Strain; Insulin-Dependent Diabetes Mellitus; Knock-out; Link; Maps; Mediating; Methods; Modeling; Mouse Strains; Mus; Mutagenesis; Names; Non obese; Other Genetics; Pathway interactions; Phenotype; Predisposition; Proteins; Regulatory T-Lymphocyte; Reporting; Resistance; Rodent Model; Role; Scheme; Serum; Susceptibility Gene; Technology; Testing; Therapeutic; Untranslated RNA; Variant; base; cell type; design; diabetic; diabetogenic; embryonic stem cell; genetic approach; genome wide association study; human disease; insight; mouse model; novel; nuclease; trait; zinc finger nuclease

PROJECT SUMMARY More than 30 autoimmune type 1 diabetes (T1D) susceptibility loci (termed Idd) have been identified in the nonobese diabetic (NOD) mouse, a spontaneous animal model for the human disease. Among those, the Idd9.3 locus has been mapped to a 1.22 Mb region on Chromosome 4 containing 17 protein-coding and 12 non-coding genes, of which Tnfrsf9 (encoding CD137) is the top candidate. The C57BL/10 (B10)-derived Idd9.3 confers T1D resistance, whereas the NOD-derived interval contributes to disease development. Due to the tight linkage of the region, congenic strains that can be used to further dissect the Idd9.3 locus have not been made available to more precisely determine if Tnfrsf9 is the underlying gene. Human genome wide association studies have linked TNFRSF9 to several autoimmune diseases, and CD137 functionally interacts in immune pathways with known human T1D genes (e.g., TNFAIP3). Thus, it is important to further study the role of CD137 in T1D. It has been previously reported that NOD-derived CD137 is hypofunctional compared to B10 CD137. On the other hand, we demonstrate that CD137-deficient NOD mice, generated by zinc-finger nuclease (ZFN) mediated mutagenesis, are resistant to T1D. We further show here that CD137 expression in CD4 and CD8 T-cells respectively suppresses and promotes T1D development in NOD mice, but CD137 deficiency dominantly affects CD8 T-cells leading to disease protection. Thus, T1D development in NOD mice carrying different alleles of Tnfrsf9 (B10, NOD, or the null allele) is influenced by the combined effects of its distinct functions in different cell types. A better approach is needed to conclusively determine if Tnfrsf9 is the Idd9.3 gene and to compare the function of NOD and B10 alleles in CD4 versus CD8 T-cells. The ability of the ZFN technology to specifically knock out a gene in mouse strains lacking germ-line transmittable embryonic stem cells has also allowed us to target Tnfrsf9 in the NOD mice congenic for the B10 Idd9.3 region. Our goal in this application is to definitively determine if Tnfrsf9 is the Idd9.3 underlying gene by establishing a pair of genetically identical strains where only one parental allele of Tnfrsf9 (NOD or B10) is expressed, and to further examine whether allelic variation of CD137 changes its dominant mode of action in T1D.

项目摘要 已经确定 非肥胖糖尿病（NOD）小鼠，一种自发性动物模型。其中， IDD9.3位点已映射到4个染色体上的1.22 MB区域，其中包含17个蛋白质编码和12个 非编码基因，其中TNFRSF9（编码CD137）是顶级候选者。 C57BL/10（B10）衍生 IDD9.3赋予了T1D抗性，而点头衍生的间隔则有助于疾病的发展。由于 该地区的紧密联系，可用于进一步剖析IDD9.3基因座的先天性菌株尚未 可以更精确地确定TNFRSF9是否是基因。人类基因组宽 协会研究将TNFRSF9与几种自身免疫性疾病联系起来，CD137在功能上相互作用 在具有已知人类T1D基因的免疫途径中（例如TNFAIP3）。因此，重要的是要进一步研究 CD137在T1D中的作用。以前已经报道了点点衍生的CD137与 B10 CD137。另一方面，我们证明了CD137缺陷的点头小鼠，由锌指生成 核酸酶（ZFN）介导的诱变对T1D具有抗性。我们在这里进一步显示CD137在 CD4和CD8 T细胞分别抑制并促进了NOD小鼠的T1D发育，但是CD137 缺乏症主要影响CD8 T细胞，导致疾病保护。因此，点头小鼠的T1D发育 携带TNFRSF9的不同等位基因（B10，点头或无效等位基因）受其综合作用的影响 不同的单元格类型中的不同功能。需要一种更好的方法来最终确定TNFRSF9是否是 IDD9.3基因并比较CD4与CD8 T细胞中NOD和B10等位基因的功能。能力 ZFN技术要专门拆除缺乏可传输胚胎的种系菌株中的小鼠菌株基因 干细胞还使我们能够针对B10 IDD9.3区域的NOD小鼠中的TNFRSF9靶向。我们的目标 在此应用中，要明确确定TNFRSF9是否是IDD9.3潜在的基因 在遗传上相同的菌株，其中仅表示TNFRSF9（点头或B10）的父母等位基因，并进一步 检查CD137的等位基因变化是否改变了其在T1D中的主要作用方式。