Role and regulation of the splicing factor ASF/SF2 in human T lymphocyte physiolo

剪接因子ASF/SF2在人T淋巴细胞生理中的作用及调控

• 批准号: 8240884
• 负责人: Vaishali Moulton
• 金额: $ 12.41万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8240884
• 关键词: Activities of Daily Living; Address; Affect; Alternative Splicing; American; Arginine; Arthritis; Autoantibodies; Autoimmune Diseases; B-Lymphocytes; Biochemical; Biological Markers; Brain; CD3 Antigens; CD44 gene; CREM protein; Cause of Death; Cell Adhesion Molecules; Cell Line; Cell physiology; Cellular Immunology; Characteristics; Child; Chronic Disease; Complex; Defect; Disease; Disease Management; Disease Marker; Disease remission; Environment; Event; Exhibits; Family member; Functional disorder; Gene Expression Regulation; Gene Silencing; Generations; Genes; Goals; Half-Life; Human; IL2 gene; Immune; Immunotherapeutic agent; Individual; Infection; Infiltration; Israel; Kidney; Longitudinal Studies; Lymphocyte Function; Medical center; Messenger RNA; Methods; Molecular; Molecular Immunology; Organ; Pain; Pathogenesis; Pathology; Patients; Phase; Phosphorylation; Physiological; Physiology; Production; Protein Binding; Protein Isoforms; Protein Splicing; Proteins; Publishing; RNA Splicing; Regulation; Relapse; Research; Resources; Role; Serine; Signal Transduction; Signaling Molecule; System; Systemic Lupus Erythematosus; T-Cell Activation; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic immunosuppression; Transducers; Variant; Woman; base; body system; child bearing; cytokine; design; medical schools; migration; molecular marker; novel; peripheral blood; programs; research study; skills; skin disorder; transcription factor

DESCRIPTION (provided by applicant): This K01 application is designed to prepare the applicant with the skills necessary to establish an independent research program on the study of systemic lupus erythematosus (SLE). SLE is an autoimmune disease of unknown cause which mainly afflicts women in their child bearing years and affects multiple organ systems of the body. T cell dysfunction due to abnormally altered intracellular signaling events is thought to be central in the pathogenesis of this disease. A critical signaling molecule known as the CD3 zeta chain is expressed in abnormally low amounts in the T cells of SLE patients, and this defect contributes to the poor functional capacity of the SLE T cells. Expression of an abnormal alternatively spliced (AS) form of the CD3 zeta mRNA with poor half life is a contributor to the reduced expression of zeta chain in SLE T cells. Aberrant splicing is a common feature observed in SLE and involves other critical genes involved in T cell signaling, migration, and cytokine production and the regulation of these genes has not been characterized. To understand the regulation of the CD3 zeta alternative splicing, the applicant used a discovery approach and identified several proteins binding to the CD3 zeta mRNA. Among these proteins, a promising candidate is the serine arginine (SR) family member Alternative Splicing Factor/Splicing Factor 2 (ASF/SF2). The applicant published that in T cells from healthy individuals, ASF/SF2 regulates CD3 zeta chain expression by suppressing production of the AS isoform. Examination of T cells from SLE patients revealed that patients had reduced expression of ASF/SF2 suggesting that aberrant ASF/SF2 expression may contribute to their defective T cell function. Additionally, the ASF/SF2 expression levels appeared to inversely correlate with the disease activity of SLE patients. The applicant's preliminary studies have shown a novel role for the splicing regulator ASF/SF2 in T cell physiology with a potential role in SLE pathophysiology. Based on the preliminary evidence, the hypothesis of this project is that ASF/SF2 regulates human T cell function and may represent a contributor to the SLE T cell defect. Toward the hypothesis, this project is aimed to understand the role of ASF/SF2 in human T lymphocytes and to determine whether ASF/SF2 may represent a disease marker in SLE. Methods will include biochemical analyses of human T cells and the development of a cell line to enable controlled expression of ASF/SF2. The specific aims of the project are 1) To determine whether TCR stimulation regulates ASF/SF2 expression and activity in human T lymphocytes 2) To determine how ASF/SF2 regulates T cell function and 3) To determine whether ASF/SF2 expression levels in SLE T cells may represent a disease marker. To accomplish these aims, the applicant will need to expand her technical and intellectual skills in cellular and molecular immunology and become proficient in the design, interpretation and analysis of studies in SLE patients. The research environment at Beth Israel Deaconess Medical Center and the other affiliates of the Harvard Medical School system will provide the applicant with the resources to reach her goals within 5 years. PUBLIC HEALTH RELEVANCE: Systemic lupus erythematosus is a chronic disease affecting over 1 million Americans. It afflicts mainly women in their child-bearing years and causes painful arthritis, with potentially fatal complications in the kidneys. The proposed project seeks to understand the potential role of an important protein ASF/SF2 in the T cells of SLE patients to assess its usefulness in disease management.

描述（由申请人提供）：此K01申请旨在为申请人准备建立有关系统性狼疮红斑eRythematosus（SLE）的独立研究计划所需的技能。 SLE是一种不明原因的自身免疫性疾病，主要使妇女在子女身上折磨，并影响人体的多个器官系统。由于异常改变的细胞内信号传导事件引起的T细胞功能障碍被认为是该疾病发病机理的核心。在SLE患者的T细胞中，称为CD3 Zeta链的临界信号分子被称为CD3 ZETA链，这种缺陷有助于SLE T细胞的功能能力较差。异常的表达脱粒（AS）的CD3 Zeta mRNA具有较差的半寿命的表达是Zeta链在SLE T细胞中的表达降低的原因。异常剪接是在SLE中观察到的常见特征，涉及与T细胞信号传导，迁移和细胞因子产生的其他关键基因，并且这些基因的调节尚未被表征。为了了解CD3 Zeta替代剪接的调节，申请人使用了发现方法，并鉴定了几种与CD3 Zeta mRNA结合的蛋白质。在这些蛋白质中，有希望的候选者是丝氨酸精氨酸（SR）家族成员替代剪接因子/剪接因子2（ASF/SF2）。申请人发表了在健康个体的T细胞中，ASF/SF2通过抑制AS同工型的产生来调节CD3 Zeta链表达。对SLE患者的T细胞的检查表明，患者的ASF/SF2表达降低，这表明异常ASF/SF2表达可能有助于其缺陷的T细胞功能。此外，ASF/SF2表达水平似乎与SLE患者的疾病活性成反比。申请人的初步研究表明，剪接调节剂ASF/SF2在T细胞生理学中具有新颖的作用 SLE病理生理学。基于初步证据，该项目的假设是ASF/SF2调节人类T细胞功能，可能代表SLE T细胞缺陷的贡献者。为了假设，该项目的目的是了解ASF/SF2在人类T淋巴细胞中的作用，并确定ASF/SF2是否可以代表SLE中的疾病标志物。方法将包括对人T细胞的生化分析和细胞系的发展，以启用ASF/SF2的控制表达。该项目的具体目的是1）确定人类T淋巴细胞中TCR刺激是否调节ASF/SF2表达和活性2）确定ASF/SF2如何调节T细胞功能和3）确定SLE T细胞中ASF/SF2表达水平是否可以代表疾病标记。为了实现这些目标，申请人将需要扩大她在细胞和分子免疫学方面的技术和智力技能，并精通SLE患者研究的设计，解释和分析。贝丝以色列执事医疗中心和哈佛医学院系统的其他分支机构的研究环境将为申请人提供5年内实现其目标的资源。 公共卫生相关性：全身性红斑狼疮是一种慢性疾病，影响了100万美国人。它主要使妇女在育儿年中受苦，并引起疼痛关节炎，并可能在肾脏中发生致命并发症。拟议的项目旨在了解重要的蛋白ASF/SF2在SLE患者T细胞中的潜在作用，以评估其在疾病管理中的有用性。