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: 9163440
• 负责人: Yi-Guang Chen
• 金额: $ 22.8万
• 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 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9163440
• 关键词: Affect; Alleles; Animal Model; Autoimmune Diseases; Autoimmune Process; Backcrossings; Biological Models; Breeding; CD4 Positive T Lymphocytes; CD8B1 gene; Candidate Disease Gene; Chromosome Mapping; Chromosomes, Human, Pair 4; Code; Complex; 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; T-Lymphocyte; Technology; Testing; Therapeutic; Untranslated RNA; Variant; base; cell type; congenic; 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 在糖尿病患者中已经鉴定了30多个自身免疫性1型糖尿病（T1 D）易感基因座（称为Idd）。 非肥胖糖尿病（NOD）小鼠，一种人类疾病的自发动物模型。其中， Idd9.3基因定位于4号染色体上1.22Mb的区域，包含17个蛋白编码区和12个蛋白编码区。 非编码基因，其中Tnfrsf 9（编码CD 137）是最佳候选者。C57 BL/10（B10）衍生型 Idd9.3赋予T1 D抗性，而NOD衍生的间隔有助于疾病发展。由于 该区域的紧密连锁，可用于进一步剖析Idd9.3基因座的同类菌株没有 可以更精确地确定Tnfrsf 9是否是潜在的基因。全人类基因组 相关研究已经将TNFRSF 9与几种自身免疫性疾病联系起来，并且CD 137在功能上相互作用， 在具有已知人T1 D基因的免疫途径中（例如，TNFAIP3）。因此，有必要进一步研究 CD 137在T1 D中的作用先前已经报道，NOD衍生的CD 137与CD 137相比功能低下。 B10 CD137。另一方面，我们证明了由锌指产生的CD 137缺陷型NOD小鼠 核酸酶（ZFN）介导的诱变，对T1 D具有抗性。我们进一步表明，CD 137表达在 CD 4和CD 8 T细胞分别抑制和促进NOD小鼠的T1 D发展，但CD 137 缺乏主要影响CD 8 T细胞，导致疾病保护。因此，NOD小鼠中的T1 D发展 携带Tnfrsf 9的不同等位基因（B10、NOD或无效等位基因）的个体受到其 在不同的细胞类型中有不同的功能。需要一种更好的方法来最终确定Tnfrsf 9是否是 Idd9.3基因，并比较NOD和B10等位基因在CD 4与CD 8 T细胞中的功能。的能力 ZFN技术特异性敲除缺乏生殖系可传播胚胎的小鼠品系中的基因 干细胞也使我们能够在与B10Idd9.3区域同源的NOD小鼠中靶向Tnfrsf 9。我们的目标 在本申请中的目的是通过建立一对Tnfrsf 9基因，确定Tnfrsf 9是否是Idd9.3潜在基因。 遗传上相同的菌株，其中仅表达Tnfrsf 9的一个亲本等位基因（NOD或B10），并且进一步 检查CD 137的等位基因变异是否改变了其在T1 D中的主导作用模式。