Mechanisms of T Cell IFNy and IL-17 Production in Juvenile Idiopathic Arthritis

项目摘要/摘要 幼年特发性关节炎(JIA)是一种以慢性关节为特征的儿童自身免疫性关节炎 发炎。辅助性T细胞1型(Th1)、Th17和病理性Th17.1细胞及炎性细胞因子 由这些细胞产生的干扰素γ和白细胞介素17(IL-17)与JIA有关 发病机制。一个主要的知识空白是了解这些炎性Th细胞和细胞因子是如何 发展。该提案概述了一项研究和培训计划，重点研究IL-17和双重干扰素γ-IL-1如何- 17产生细胞在JIA Th1分化过程中发育不当。这些研究的一个目标是确定 可用于改进诊断和治疗决策的生物因素。调查员进行了 研究描述：1)经过Th1分化的多关节JIA循环细胞产生高 IL-17和干扰素γ水平及双重干扰素γ-IL-17产生细胞2)1例少见功能丧失的强直性脊柱炎患者 GATA3突变表现出这种表型的夸大形式，3)额外的JIA患者携带罕见的 对Th1分化至关重要的基因中的蛋白质编码突变。该提案的中心假设是 贾氏Th1分化不当产生IL-17和Th1.17细胞罕见基因突变导致 这种表型。提出的具体目标验证了这一假设。目标1确定新基因的作用 乙型病毒性肝炎患者Th1分化过程中产生IL-17、干扰素γ和Th1.17细胞的突变。目标2 识别导致IL-17和Th1.17细胞产生的细胞因子和STAT信号通路 在多关节JIA Th1分化过程中。这项建议涉及对人类细胞的翻译研究，使用 先进的细胞学、分子生物学和下一代测序。这项提议的一个主要焦点是 支持帕特里克医生作为内科科学家的发展。她的职业目标是研究JIA的发病机制 在基础研究计划中，确定产生新的治疗靶点和改善的生物因素 诊断和治疗决策。她将通过职业目标实现这一目标，成为一名 免疫学专家，获得免疫技术方面的高级专业知识，熟练使用和 分析下一代测序，并获得成为独立首席研究员的技能。 该提案的研究环境非常突出。帕特里克医生有完整的部门和机构 支持她的研究项目的发展。她的导师都是免疫学家，在 有计划的先进技术。范德比尔特拥有一流的设施和共享的资源来进行这些方面的培训 技巧。她的指导团队包括免疫学、基因调控和风湿病方面的专家 指导她的独立研究项目的发展。这个K08将支持数据的生成和 导致出版物提供对JIA的发病机制和遗传学的洞察，并支持成功的 R01应用程序。

PROJECT SUMMARY/ABSTRACT Juvenile idiopathic arthritis (JIA) is an autoimmune arthritis in children characterized by chronic joint inflammation. T helper type 1 (Th1), Th17, and pathologic Th17.1 cells and the inflammatory cytokines produced by these cells, interferon gamma (IFNγ) and interleukin-17 (IL-17), are implicated in JIA pathogenesis. A major knowledge gap is to understand how these inflammatory Th cells and cytokines develop. This proposal outlines a research and training plan focused on studying how IL-17 and dual IFNγ-IL- 17 producing cells inappropriately develop during JIA Th1 differentiation. A goal for these studies is to identify biologic factors that can be used to improve diagnostic and therapeutic decisions. The investigator conducted studies that described: 1) polyarticular JIA circulating cells that underwent Th1 differentiation produced high levels of IL-17 and IFNγ and dual IFNγ-IL-17 producing cells, 2) a JIA patient with a rare loss-of-function GATA3 mutation exhibited an exaggerated form of this phenotype, and 3) additional JIA patients carry rare protein-coding mutations in genes important for Th1 differentiation. The proposal’s central hypothesis is that JIA Th1 differentiation inappropriately produces IL-17 and Th1.17 cells and rare genetic mutations contribute to this phenotype. The proposed Specific Aims test this hypothesis. Aim 1 identifies the role of novel genetic mutations from JIA patients in production of IL-17, IFNγ, and Th1.17 cells during Th1 differentiation. Aim 2 identifies the cytokine and STAT signaling pathways that lead to the production of IL-17 and Th1.17 cells during polyarticular JIA Th1 differentiation. This proposal involves translational studies in human cells using advanced cytometry, molecular biology, and next generation sequencing. A major focus of this proposal is to support Dr. Patrick’s development as a physician-scientist. Her career goal is to study the pathogenesis of JIA in a basic research program and identify biologic factors that generate novel therapeutic targets and improve diagnostic and therapeutic decisions. She will accomplish this goal through career aims to become an immunology expert, gain advanced expertise in immunologic techniques, establish proficiency in the use and analysis of next-generation sequencing, and acquire skillsets to become an independent principal investigator. The research environment for the proposal is outstanding. Dr. Patrick has full departmental and institutional support for the development of her research program. Her mentors are immunologists with expertise in the planned advanced techniques. Vanderbilt has excellent facilities and shared resources for training in these techniques. Her mentoring team includes experts in immunology, gene regulation, and rheumatologic disease to guide development of her independent research program. This K08 will support the generation of data and lead to publications providing insight into the pathogenesis and genetics of JIA and in support of a successful R01 application.