IRF5-TNPO3 locus: Inclusion of TNPO3 as a unique regulator of IRF5 expression an

IRF5-TNPO3 基因座：将 TNPO3 作为 IRF5 表达的独特调节因子纳入其中

• 批准号: 8664081
• 负责人: Betsy J Barnes
• 金额: $ 20.99万
• 依托单位: RBHS-NEW JERSEY MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8664081
• 关键词: Affect; Alternative Splicing; American; Arginine; Autoantibodies; Autoimmune Diseases; B-Lymphocytes; Blood Cells; Cell Nucleus; Cells; Chromosomes, Human, Pair 7; Clinical; Complex; Data; Diabetes Mellitus; Disease; Face; Family; Flow Cytometry; Gene Expression; Genes; Genetic; Genetic Polymorphism; Genetic Risk; Genetic Variation; Genotype; Goals; Haplotypes; Health; Heart Diseases; Heterogeneity; Human Genetics; IL6 gene; Image; Immune; Immune response; Immunity; Immunoblot Analysis; Immunologics; Immunoprecipitation; Importins; Interferons; Interleukin-10; Interleukin-12; Joints; Karyopherins; Knowledge; Lupus; Malignant Neoplasms; Mediating; Nuclear; Nuclear Antigens; Nuclear Import; Nuclear Localization Signal; Nuclear Pore Complex Proteins; Pathogenesis; Pathology; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Play; Population; Predisposition; Prevalence; Process; Production; Protein Import; Proteins; Publishing; RNA Splicing; Regulation; Research Proposals; Reverse Transcriptase Polymerase Chain Reaction; Rheumatoid Arthritis; Risk; Role; Sampling; Self Tolerance; Serine; Serum; Single Nucleotide Polymorphism; Systemic Lupus Erythematosus; Technology; Testing; Transcript; Tumor Necrosis Factor-alpha; United States National Institutes of Health; Validation; Work; base; chemokine; chemokine receptor; cohort; cytokine; design; disease phenotype; disorder risk; drug discovery; fluorescence imaging; genetic variant; genome wide association study; human ARMET protein; insight; lymphoblastoid cell line; member; next generation sequencing; novel; overexpression; protein expression; protein function; prototype; receptor; receptor expression; statistics; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): Project Summary The NIH estimates up to 23.5 million Americans suffer from autoimmune diseases and that the prevalence is rising; this statistic is higher than the number of Americans having cancer (9 million) or heart disease (22 million). Systemic lupus erythematosus (SLE) is the third most frequent autoimmune disease affecting approximately 1 in 2000 people. It is the prototype of complex autoimmune diseases and is characterized by extreme breakdown of self-tolerance resulting in multiple immunologic abnormalities. Cures are not yet available for SLE and patients face a lifetime of illness and treatment. Given the clinical heterogeneity of this disease, combined with limited knowledge of pathogenesis, designing new treatment options has been difficult. Recent findings from joint linkage and genome-wide association studies (GWAS) have provided significant insight into the genes and pathways contributing to disease pathogenesis. SLE is highly heritable with a strong genetic component that includes the interferon regulatory factor 5 (IRF5)-transportin 3 (TNPO3) region. Genetic variants in the IRF5-TNPO3 locus have been associated with SLE risk in every published study testing this region and in every ethnic population. Although some effort has recently gone into studying the functional consequence of genetic variants in IRF5, few studies have included an analysis of TNPO3 expression and function in SLE. The primary goal of this research proposal is to determine how the IRF5-TNPO3 locus alters SLE disease risk by experimentally bridging the gap(s) in knowledge between identification of this causal locus and its actual functional influence on SLE risk. IRF5 encodes a transcription factor central to the innate and adaptive immune response through its regulation of key processes including cytokine production, chemokine and chemokine receptor expression, and autoantibody production. TNPO3 is a member of the importin family and encodes a nuclear import receptor that mediates nuclear entry of serine/arginine (SR)-rich proteins such as splicing factors SFRS1 and SFRS2. Based on recent experimental findings from our lab, we hypothesize that the genetic risk coming from the IRF5-TNPO3 locus is not solely from IRF5 and directly involves TNPO3 function via its ability to regulate novel mechanisms controlling IRF5 alternative splicing and nuclear localization. This hypothesis will be tested in the following Specific Aims: 1) Determine the level of IRF5 and TNPO3 transcript and protein expression in immune cells from healthy donors and SLE patients carrying the risk and non-risk haplotypes at IRF5-TNPO3 SNPs, 2) Determine whether TNPO3 controls IRF5 transcript expression through its ability to regulate the import of SR proteins to the nucleus, and 3) Determine whether IRF5, TNPO3 and NUP62 differentially interact in genotyped healthy donor and SLE immune cells resulting in altered IRF5 nuclear localization. Results from this study will bridge the gap between disease- associated genetic variation, gene expression, protein function, and disease phenotype, and identify novel mechanisms that can be therapeutically targeted to regulate IRF5 expression and activation in SLE.

描述（由申请人提供）： 项目摘要 NIH 估计多达 2350 万美国人患有自身免疫性疾病，而且患病率正在上升；这一统计数字高于患有癌症（900 万）或心脏病（2200 万）的美国人人数。系统性红斑狼疮 (SLE) 是第三大最常见的自身免疫性疾病，约有 2000 人中就有 1 人受到影响。它是复杂自身免疫性疾病的原型，其特征是自我耐受性的极度崩溃，导致多种免疫异常。 SLE 尚无治愈方法，患者面临终生的疾病和治疗。鉴于这种疾病的临床异质性，加上对发病机制的了解有限，设计新的治疗方案一直很困难。联合连锁和全基因组关联研究（GWAS）的最新发现为了解疾病发病机制的基因和途径提供了重要的见解。 SLE 具有高度遗传性，具有强大的遗传成分，其中包括干扰素调节因子 5 (IRF5)-转运蛋白 3 (TNPO3) 区域。在测试该地区和每个种族人群的每一项已发表的研究中，IRF5-TNPO3 位点的遗传变异都与 SLE 风险相关。尽管最近在研究 IRF5 遗传变异的功能后果方面做出了一些努力，但很少有研究包括对 SLE 中 TNPO3 表达和功能的分析。本研究计划的主要目标是通过实验弥合该因果基因座的识别与其对 SLE 风险的实际功能影响之间的知识差距，确定 IRF5-TNPO3 基因座如何改变 SLE 疾病风险。 IRF5 通过调节细胞因子产生、趋化因子和趋化因子受体表达以及自身抗体产生等关键过程，编码对先天性和适应性免疫反应至关重要的转录因子。 TNPO3 是输入蛋白家族的成员，编码一种核输入受体，可介导富含丝氨酸/精氨酸 (SR) 的蛋白质（例如剪接因子 SFRS1 和 SFRS2）进入核。根据我们实验室最近的实验结果，我们假设来自 IRF5-TNPO3 基因座的遗传风险不仅仅来自 IRF5，还通过其调节控制 IRF5 选择性剪接和核定位的新机制的能力直接涉及 TNPO3 功能。该假设将在以下具体目标中进行测试：1) 确定来自携带 IRF5-TNPO3 SNP 风险和非风险单倍型的健康捐赠者和 SLE 患者的免疫细胞中 IRF5 和 TNPO3 转录物和蛋白质表达的水平，2) 确定TNPO3 是否通过其调节 SR 蛋白向细胞核输入的能力来控制 IRF5 转录本表达，以及 3) 确定 IRF5、TNPO3 和 NUP62 是否基因分型的健康供体和 SLE 免疫细胞之间存在差异性相互作用，导致 IRF5 核定位发生改变。这项研究的结果将弥合疾病相关遗传变异、基因表达、蛋白质功能和疾病表型之间的差距，并确定可在治疗上靶向调节 SLE 中 IRF5 表达和激活的新机制。