Mutant PTPN22 in autoimmune mice

自身免疫小鼠中的突变 PTPN22

• 批准号: 9095202
• 负责人: LINDA A SHERMAN
• 金额: $ 14.44万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9095202
• 关键词: Affect; Alleles; American; Autoimmune Diabetes; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Biochemical; CRISPR/Cas technology; Cells; Cytotoxic T-Lymphocyte-Associated Protein 4; Development; Diabetes Mellitus; Disease; Disease Progression; Embryo; Experimental Models; Female; Frameshift Mutation; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Germ Lines; Germ-Line Mutation; Goals; Health; Homeostasis; Human; Immune; Inbred NOD Mice; Inbreeding; Incidence; Infiltration; Insulin; Insulin-Dependent Diabetes Mellitus; Lead; Link; Lymphocyte Function; Lymphoid Cell; Modeling; Mus; Mutate; Mutation; Myeloid Cells; Non obese; Orthologous Gene; PTPN22 gene; Pathway interactions; Phosphoric Monoester Hydrolases; Population; Prevention therapy; Protein Tyrosine Phosphatase; Proteins; Publications; Relative Risks; Rheumatoid Arthritis; Role; Single Nucleotide Polymorphism; Systemic Lupus Erythematosus; Systemic Scleroderma; Technology; Time; Variant; base; cell type; diabetic; diabetic patient; genetic variant; genome wide association study; immune function; innovation; insertion/deletion mutation; insulin dependent diabetes mellitus onset; islet; male; mouse model; mutant; novel therapeutics; prevent; protein expression; transmission process

 DESCRIPTION (provided by applicant): Autoimmunity is a multigenic disease that generally involves the presence of the pro- autoimmune allele at numerous genetic loci. There is a strong association between a single nucleotide polymorphism (SNP) in the PEST-domain containing intracellular protein tyrosine phosphatase, PTPN22 R620W, and numerous autoimmune diseases, including systemic lupus erythematosus, systemic sclerosis, rheumatoid arthritis, Graves diseases and Type 1 Diabetes. It is not understood how this allele contributes to these diseases. Type 1 diabetes (T1D) is increased in incidence by 2-4 fold when this allele is present. We propose to use the non-obese diabetic (NOD) mouse model of spontaneous type 1 diabetes (T1D) to determine how the mouse orthologue of R620W, R619W, contributes to disease. This model is ideal as many of the genes identified by genome- wide association studies in diabetic patients are the same or on the same pathway as those that contribute to T1D in NOD mice, including MHC, insulin, CTLA-4, and the IL- 2R pathway. The presence of the pro-autoimmune alleles may be critical to assess the impact of the R619W allele on T1D. We have successfully used the Crispr-Cas9 technology to mutate NOD fertilized embryos and produce 2 independent lines of NOD mice that express the R619W variant. We have also obtained several lines that incorporated insertions or deletions (indels) that caused frameshift mutations in this gene. We have ascertained all mutations are germline transmitted, and now propose to determine how they affect PTPN22 expression, immune cell development and homeostasis, and most importantly, disease progression. To this end we will pursue 2 Specific Aims. Aim1 will assess the level of expression of the mutant PTPN22 in various immune cell types from the independently derived NOD murine lines. The primary goal of Aim 2 is to determine whether the R619W and the indel mutants alter the incidence and progression of T1D. We will also assess its effect on immune cell development, function and homeostasis in NOD mice.

 描述（由申请人提供）：自身免疫是一种多基因疾病，通常涉及在许多遗传位点存在促自身免疫等位基因。含有细胞内蛋白酪氨酸磷酸酶 PTPN22 R620W 的 PEST 结构域中的单核苷酸多态性 (SNP) 与许多自身免疫性疾病（包括系统性红斑狼疮、系统性硬化症、类风湿性关节炎、格雷夫斯病和 1 型糖尿病）之间存在密切关联。目前尚不清楚该等位基因如何导致这些疾病。当该等位基因存在时，1 型糖尿病 (T1D) 的发病率会增加 2-4 倍。我们建议使用自发性 1 型糖尿病 (T1D) 的非肥胖糖尿病 (NOD) 小鼠模型来确定 R620W、R619W 的小鼠直系同源物如何导致疾病。该模型是理想的，因为糖尿病患者的全基因组关联研究鉴定出的许多基因与 NOD 小鼠中导致 T1D 的基因相同或位于相同的途径，包括 MHC、胰岛素、CTLA-4 和 IL-2R 途径。促自身免疫等位基因的存在对于评估 R619W 等位基因对 T1D 的影响可能至关重要。我们成功地利用 Crispr-Cas9 技术对 NOD 受精胚胎进行突变，并产生了 2 个表达 R619W 变体的独立 NOD 小鼠品系。我们还获得了几个包含导致该基因移码突变的插入或缺失（indels）的品系。我们已经确定所有突变都是种系传播的，现在建议确定它们如何影响 PTPN22 表达、免疫细胞发育和体内平衡，以及最重要的是疾病进展。为此，我们将追求 2 个具体目标。 Aim1 将评估来自独立衍生的 NOD 小鼠系的各种免疫细胞类型中突变体 PTPN22 的表达水平。目标 2 的主要目标是确定 R619W 和插入缺失突变体是否会改变 T1D 的发病率和进展。我们还将评估其对 NOD 小鼠免疫细胞发育、功能和体内平衡的影响。

项目成果

CRISPR-Cas9-Mediated Modification of the NOD Mouse Genome With Ptpn22R619W Mutation Increases Autoimmune Diabetes.

• DOI: 10.2337/db16-0061
• 发表时间: 2016-08
• 期刊: Diabetes
• 影响因子: 7.7
• 作者: Lin X;Pelletier S;Gingras S;Rigaud S;Maine CJ;Marquardt K;Dai YD;Sauer K;Rodriguez AR;Martin G;Kupriyanov S;Jiang L;Yu L;Green DR;Sherman LA
• 通讯作者: Sherman LA