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PTPN22 R619W in NOD mice

NOD 小鼠中的 PTPN22 R619W

基本信息

批准号：
9307520
负责人：
LINDA A SHERMAN
金额：
$ 76.27万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9307520
关键词：
Adoptive Transfer Affect Alleles Amino Acids Antigens Appearance Autoantibodies Autoimmune Diseases Autoimmune Process Autoimmunity Automobile Driving B-Lymphocytes Behavior Birth Bone Marrow C57BL/6 Mouse CD4 Positive T Lymphocytes CRISPR/Cas technology Candidate Disease Gene Cell physiology Cells Child Data Defect Development Diabetes Mellitus Discrimination Disease Disease susceptibility Drug Targeting Environmental Risk Factor Enzymes European Exhibits Frequencies Gene Expression Gene Expression Profiling Genes Genetic Genetic Polymorphism Goals Graves&apos Disease Haplotypes Human Immune Immune Tolerance Immune system Inbred NOD Mice Incidence Insulin Insulin Antibodies Insulin-Dependent Diabetes Mellitus Islets of Langerhans Label Lymphocyte Modeling Molecular Monitor Mus Mutate Mutation Myelogenous Myeloid Cells Non obese Orthologous Gene Pancreas Pathway interactions Penetrance Peptides Phenotype Population Predisposition Preventive therapy Production Property Protein Tyrosine Phosphatase Proteins Retroviral Vector Rheumatoid Arthritis Risk Risk Factors Role Single Nucleotide Polymorphism Structure of beta Cell of islet Structure of germinal center of lymph node Susceptibility Gene System Systemic Lupus Erythematosus T-Lymphocyte Techniques Testing Transgenic Mice Transgenic Organisms Vitiligo anergy base cell type differential expression disorder risk experimental study genetic risk factor genome editing high risk human disease insulin tolerance interest islet knock-down lymph nodes mouse model prevent reconstitution seroconversion small hairpin RNA therapy development transcriptome sequencing

项目摘要

Project Summary: Type 1 diabetes (T1D) results from a breakdown in immune tolerance that progresses to the destruction of the insulin producing beta cells of the pancreatic islets. A large number of genetic and environmental factors contribute to the incidence of disease. One of the strongest non-MHC genetic risk factors is a single nucleotide polymorphism in the gene that encodes the tyrosine phosphatase non-receptor type 22, PTPN22, that results in a single amino acid change in the protein, R620W. PTPN22 is expressed in many bone marrow derived cells, including lymphocytes and myeloid derived cells. Although R620W increases the risk of many autoimmune diseases associated with production of autoantibodies, including T1D, rheumatoid arthritis, systemic lupus erythematosus, Graves disease, vitiligo, and others, little is know about how it enhances disease. In part, this is because it functions in many different types of cells that affect the immune system. In order to produce a mouse model that can be used to identify its mechanism of action, we used Crispr/Cas9 genome editing to introduce the mouse ortholog of the pro-autoimmune allele of PTPN22 (619W) into the non- obese diabetes (NOD) mouse that spontaneously develops T1D. NOD mice harboring this mutation demonstrate accelerated production of anti-insulin antibodies and accelerated T1D of higher penetrance, which is the same phenotype exhibited by humans expressing R620W. This proposal will use the 619W NOD model to reveal the mechanism responsible for enhanced disease by pursuing the following specific aims: Aim1 will identify immune cells in which endogenous expression of 619W enhances IAA production and T1D. This entails adoptive transfer experiments to produce mice expressing the 619W mutation of PTPN22 in key immune cells. Aim 2 will explore the mechanistic basis for enhanced IAA and T1D in 619W mice. The types of immune cells identified under Aim 1 will be probed to identify genes affected in their level of expression by allelic difference at 619. Genes and pathways will be identified by either RNAseq or by analyzing gene expression at the single cell level. Candidate genes will be tested for function by altering level of expression with shRNAs delivered by retroviral vectors. Aim 3 will examine tolerance of insulin specific B lymphocytes in R619W mice. Aim 4 will use TCR transgenic mice and single cell expression analysis to examine the fate of islet antigen specific T cells as they become activated in either 619W or 619R expressing mice. Successful completion of these studies will have relevance to development of treatments that may prevent the many different autoimmune disease associated with PTPN22.

项目总结: