Immune Dysregulation in Pediatric SLE Pathogenesis

儿童 SLE 发病机制中的免疫失调

• 批准号: 9223553
• 负责人: WEN-YUAN E HSIEH
• 金额: $ 17.06万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-07 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9223553
• 关键词: Adaptive Immune System; Adult; Advisory Committees; Antibodies; Autoantigens; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Basic Science; Biological; Biological Assay; Biological Markers; Biology; Biometry; Blood donor; Cells; Chairperson; Child; Childhood; Clinical; Clinical Research; Collaborations; Colorado; Complement; Computational Biology; Cytokine Network Pathway; Cytometry; Data; Data Analyses; Development Plans; Diagnosis; Diagnostic; Disease; Environment; Event; Exhibits; Exposure to; Flare; Goals; Human; Immune; Immune signaling; Immune system; Immunology; Immunomodulators; Immunosuppression; Inflammation; Inflammatory; Innate Immune System; International; Isotopes; JAK1 gene; Knowledge; Laboratories; Lipids; Liquid substance; Longitudinal cohort; Lymphocyte Activation; Macrophage Inflammatory Proteins; Mediating; Mediator of activation protein; Mentors; Microbiology; Mission; Molecular; Monitor; Monocyte Chemoattractant Protein-1; Morbidity - disease rate; Organ; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pattern; Pediatric Hospitals; Pediatrics; Phenotype; Pilot Projects; Play; Production; Prognostic Marker; Property; Proteins; Proteomics; Public Health; Rare Earth Metals; Recurrent disease; Reproducibility; Research; Research Design; Resolution; Rheumatism; Role; Scientist; Serum; Severity of illness; Signal Pathway; Surface; System; Systemic Lupus Erythematosus; Technology; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Toll-like receptors; Training; United States National Institutes of Health; Universities; Variant; anakinra; base; career development; chemokine; cohort; cytokine; exosome; experience; extracellular vesicles; fluorophore; high dimensionality; immunoregulation; inhibitor/antagonist; innovation; knowledge base; monocyte; mortality; multidisciplinary; new therapeutic target; novel; outcome forecast; pediatric patients; peripheral blood; professor; prognostic; prospective; response; rheumatologist; single cell analysis; targeted treatment; therapeutic target; translational impact

PROJECT SUMMARY Systemic lupus erythematosus (SLE) is a multi-organ rheumatologic disease characterized by immune dysregulation and a heterogeneous disease course, with heightened disease severity in pediatric patients. Conventional clinical and laboratory parameters are not sufficiently sensitive or specific for detecting ongoing disease activity or response to specific immunomodulators, underscoring a need to better delineate the underlying immune changes that drive unpredictable flare-ups of disease activity. The lack of knowledge regarding the precise immune cellular and molecular events leading to SLE disease poses a significant hurdle in the effort to develop accurate prognostic disease biomarkers and selective therapeutic agents. Our long- term goal is to identify immune cellular and molecular mediators that could provide targets for therapeutic intervention. The objective of this proposal is to elucidate the mechanisms by which serum-circulating factors including exosomes modulate immune cellular and cytokine derangements in pediatric SLE pathogenesis. The rationale for the proposed research is that understanding these mechanisms could identify novel disease biomarkers that permit accurate prognosis and provide selective therapeutic targets. To achieve this goal, Dr. Hsieh will use a high-dimensional mass cytometry platform, which offers single-cell analysis of over 40 parameters, utilizing rare earth metal isotopes instead of fluorophores as tags bound to antibodies. In a pilot study, Dr. Hsieh established a quantitative and reproducible mass cytometry assay that allows precise monitoring of phenotypic (surface markers) and functional (cytokines/chemokines) immune derangements detectable in peripheral blood of pediatric SLE patients. The data from this study form the basis of Dr. Hsieh's central hypothesis that pediatric SLE patients share a unique multi-parametric monocyte cytokine signature, which drives disease activity and is propagated by serum factors (including exosomes) through activation of the JAK/STAT signaling pathway. Guided by strong preliminary data, this hypothesis will be tested by pursuing three specific aims: 1) Determine the association of the cytokine signature (and other immune changes) with SLE disease activity, by integrating mass cytometry immune profile data and clinical parameters from a prospective longitudinal pediatric SLE patient cohort; 2) Elucidate immunomodulatory properties of SLE serum-isolated exosomes, by evaluating the ability of diseased serum exosomes to induce the cytokine signature, and identifying exosome proteomic components essential for such activity; and 3) Determine the capacity of ruxolitinib (JAK1/2 inhibitor) to abrogate expression of the cytokine signature, by evaluating ex vivo immunosuppressive effects (including off-target effects) of ruxolitinib on immune derangements observed in pediatric SLE pathogenesis. Dr. Hsieh is an Assistant Professor at the University of Colorado Denver/Children's Hospital Colorado in the Departments of Immunology and Microbiology, and Pediatrics. As a clinical fellow, she trained in Dr. Garry Nolan's laboratory (Stanford), acknowledged as world-class in the application of mass cytometry to single-cell analysis, where she acquired expertise in this technology. Dr. Hsieh has assembled a mentoring team composed of a multidisciplinary group of scientists with research expertise in basic science immunology, particularly immune signaling alterations in autoimmunity; clinical study design and implementation; exosome biology; and biostatistics. Her primary mentor Dr. John Cambier and co-mentor Dr. V. Michael Holers are well- established scientists with collaborations focused on autoimmune disorders and the immune system. Their extensive experience with trainees will be complemented by the expertise of her scientific advisory committee, co-mentor Dr. Michael Graner, internationally known expert in the field of exosome biology, advisors/collaborators Dr. Jennifer Soep, pediatric rheumatologist with a clinical research focus on SLE, and Dr. Debashis Ghosh, Chairman of Biostatistics with expertise on high-throughput data analysis. Collectively, this team will provide an outstanding training environment that will fill critical gaps in her toolbox and knowledge base to enhance her ability to study immune derangements in pediatric SLE patients. Dr. Hsieh has created a career development plan that will prepare her for successful transition to independence, incorporating didactic and mentored training in: 1) basic science immunology; 2) clinical study design and implementation; 3) exosome biology; and 4) biostatistics and computational biology. The proposed approach is innovative, in the applicant's opinion, because it departs from the status quo by elucidating specific mechanisms underlying inflammatory perturbations in SLE (e.g., serum exosome-mediated immune modulation) and by characterizing these mechanisms as they occur in the context of multiple immune cell phenotypes, intracellular cytokine networks, and remitting/relapsing disease activity, with single-cell level resolution. The proposed research is significant because it will likely identify candidate disease biomarkers for prognostic and therapeutic purposes in SLE, and will likely delineate novel therapeutic targets derived from serum exosomes and their proteomic components. The knowledge gained will have broad translational importance through application of this systems immunology approach to other systemic rheumatologic disorders.

项目概要 系统性红斑狼疮（SLE）是一种以免疫系统损害为特征的多器官风湿病 失调和异质性病程，儿科患者的疾病严重程度更高。 传统的临床和实验室参数对于检测持续存在的情况不够敏感或特异。 疾病活动或对特定免疫调节剂的反应，强调需要更好地描述 潜在的免疫变化会导致不可预测的疾病活动突然爆发。缺乏知识 关于导致 SLE 疾病的精确免疫细胞和分子事件构成了重大障碍 努力开发准确的预后疾病生物标志物和选择性治疗药物。我们的长期 长期目标是确定可以为治疗提供靶点的免疫细胞和分子介质 干涉。该提案的目的是阐明血清循环因子的机制 包括外泌体在儿科 SLE 发病机制中调节免疫细胞和细胞因子紊乱。这 拟议研究的基本原理是了解这些机制可以识别新疾病 允许准确预后并提供选择性治疗靶点的生物标志物。 为了实现这一目标，谢博士将使用高维质谱流式分析平台，该平台提供单细胞分析 使用稀土金属同位素而不是荧光团作为结合到的标签来分析 40 多个参数 抗体。在一项试点研究中，Hsieh 博士建立了一种定量且可重复的质谱流式分析方法， 允许精确监测表型（表面标记）和功能（细胞因子/趋化因子）免疫 儿童 SLE 患者外周血中可检测到的紊乱。本研究的数据构成基础 谢博士的中心假设是，儿童系统性红斑狼疮患者拥有独特的多参数单核细胞 细胞因子特征，驱动疾病活动并通过血清因子（包括外泌体）传播 通过激活 JAK/STAT 信号通路。在强有力的初步数据的指导下，这一假设将 通过追求三个具体目标来进行测试：1）确定细胞因子特征（和其他 通过整合质谱流式细胞仪免疫特征数据和临床数据，将免疫变化与 SLE 疾病活动相关 来自前瞻性纵向儿科 SLE 患者队列的参数； 2) 阐明免疫调节作用 通过评估患病血清外泌体诱导 SLE 血清分离的外泌体的特性 细胞因子特征，并鉴定此类活性所必需的外泌体蛋白质组成分；和 3) 确定鲁索替尼（JAK1/2 抑制剂）消除细胞因子特征表达的能力，方法是 评估鲁索替尼对免疫的离体免疫抑制作用（包括脱靶作用） 儿童 SLE 发病机制中观察到的紊乱。 谢博士是科罗拉多大学丹佛分校/科罗拉多州儿童医院的助理教授 免疫学和微生物学系以及儿科。作为一名临床研究员，她接受了加里博士的培训 诺兰实验室（斯坦福），在单细胞质谱应用方面被公认为世界一流 分析，她在那里获得了这项技术的专业知识。谢博士组建了导师团队 由具有基础科学免疫学研究专业知识的多学科科学家小组组成， 特别是自身免疫中的免疫信号改变；临床研究设计和实施；外泌体 生物学;和生物统计学。她的主要导师 John Cambier 博士和副导师 V. Michael Holers 博士非常好—— 建立了专注于自身免疫性疾病和免疫系统的科学家合作。他们的 她的科学咨询委员会的专业知识将补充与受训者的丰富经验， 共同导师Michael Graner博士，外泌体生物学领域国际知名专家， 顾问/合作者 Jennifer Soep 博士，儿科风湿病学家，专注于 SLE 临床研究，以及 Debashis Ghosh 博士，生物统计学主席，拥有高通量数据分析方面的专业知识。总的来说， 该团队将提供出色的培训环境，填补她工具箱中的关键空白， 知识库，以提高她研究儿童 SLE 患者免疫紊乱的能力。谢博士有 制定了职业发展计划，为她成功过渡到独立做好准备， 将教学和指导培训纳入： 1) 基础科学免疫学； 2) 临床研究设计和 执行; 3）外泌体生物学； 4）生物统计学和计算生物学。 申请人认为，所提出的方法具有创新性，因为它通过以下方式偏离了现状： 阐明 SLE 炎症扰动的具体机制（例如血清外泌体介导的 免疫调节）并通过表征这些机制，因为它们发生在多重免疫的背景下 单细胞水平的细胞表型、细胞内细胞因子网络和缓解/复发疾病活动 解决。拟议的研究意义重大，因为它可能会确定候选疾病生物标志物 SLE 的预后和治疗目的，并可能描绘出源自 SLE 的新治疗靶点 血清外泌体及其蛋白质组成分。所获得的知识将具有广泛的转化 通过将该系统免疫学方法应用于其他系统性风湿病的重要性 失调。