Immune Dysregulation in Pediatric SLE Pathogenesis

儿童 SLE 发病机制中的免疫失调

• 批准号: 9902184
• 负责人: WEN-YUAN E HSIEH
• 金额: $ 16.97万
• 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/9902184
• 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; CCL2 gene; 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; Morbidity - disease rate; Organ; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pattern; Pediatric Hospitals; Pediatric cohort; 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.

项目摘要 系统性红斑狼疮（SLE）是一种多器官风湿性疾病， 失调和异质性病程，在儿科患者中具有升高的疾病严重程度。 常规的临床和实验室参数对于检测持续的 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，驱动不可预测的疾病活动的爆发。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一个重大障碍 致力于开发准确的预后疾病生物标志物和选择性治疗剂。我们长久以来- 长期目标是鉴定免疫细胞和分子介质， 干预这项建议的目的是阐明血清循环因子的机制， 包括外泌体调节儿童SLE发病机制中的免疫细胞和细胞因子紊乱。的 这项研究的基本原理是，了解这些机制可以识别新的疾病， 生物标志物允许准确的预后并提供选择性治疗靶点。 为了实现这一目标，Hsieh博士将使用一个高维的质谱细胞仪平台，该平台提供单细胞 分析超过40个参数，利用稀土金属同位素代替荧光团作为结合到 抗体的在一项初步研究中，Hsieh博士建立了一种定量和可重复的质谱细胞计数法， 允许精确监测表型（表面标志物）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。这项研究的数据构成了 谢博士的中心假设，即儿童SLE患者共享一个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（包括外来体）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进行测试：1）确定细胞因子标签（和其他细胞因子标签）与细胞因子之间的关联。 免疫变化）与SLE疾病活动性的关系，通过整合质谱免疫谱数据和临床 来自前瞻性纵向儿科SLE患者队列的参数; 2）阐明免疫调节 SLE血清分离的外泌体的性质，通过评估患病血清外泌体诱导 细胞因子特征，并鉴定这种活性所必需的外泌体蛋白质组组分;以及3） 通过以下方式确定鲁索替尼（JAK 1/2抑制剂）消除细胞因子标签表达的能力： 评价鲁索利替尼对免疫细胞的离体免疫抑制作用（包括脱靶作用） 在儿童SLE发病机制中观察到的紊乱。 博士谢是科罗拉多大学丹佛分校/科罗拉多儿童医院的助理教授， 免疫学、微生物学和儿科学系。作为一名临床研究员，她在加里医生 诺兰的实验室（斯坦福大学），被公认为世界一流的应用大量细胞计数单细胞 分析，在那里她获得了这项技术的专业知识。谢博士组建了一个指导小组 由具有基础科学免疫学研究专长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;生物统计学。她的主要导师约翰·坎比尔博士和共同导师V·迈克尔·霍勒斯博士都很好- 建立科学家与合作的重点是自身免疫性疾病和免疫系统。他们的 她的科学咨询委员会的专门知识将补充她对受训人员的广泛经验， 共同导师Michael Graner博士，国际知名的外泌体生物学领域专家， 顾问/合作者Jennifer Soep博士，儿科风湿病学家，专注于SLE的临床研究， 博士Debashis Ghosh，生物统计学主席，擅长高通量数据分析。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， 知识基础，以提高她的能力，研究免疫紊乱的儿童系统性红斑狼疮患者。谢博士 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 实施; 3）外泌体生物学;以及4）生物统计学和计算生物学。 申请人认为，拟议的办法是创新的，因为它通过以下方式改变了现状： 阐明了SLE中炎性扰动的具体机制（例如，血清外泌体介导 免疫调节），并通过表征这些机制，因为它们发生在多种免疫调节的背景下， 细胞表型、细胞内细胞因子网络和缓解/复发性疾病活动，单细胞水平 分辨率这项拟议中的研究意义重大，因为它可能会确定候选疾病生物标志物， 的预后和治疗目的，并可能描绘新的治疗靶点， 血清外泌体及其蛋白质组成分。获得的知识将有广泛的翻译 通过将这种系统免疫学方法应用于其他系统性风湿病 紊乱