Immune Dysregulation in Pediatric SLE Pathogenesis

儿童 SLE 发病机制中的免疫失调

• 批准号: 10155413
• 负责人: WEN-YUAN E HSIEH
• 金额: $ 16.93万
• 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 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10155413
• 关键词: Adaptive Immune System; Adult; Advisory Committees; Antibodies; Autoantigens; Autoimmune; Autoimmune Diseases; 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; Prognosis; 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; 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 全身性红斑狼疮（SLE）是一种以免疫为特征的多器官风湿病 失调和异质性疾病病程，儿科患者的疾病严重程度升高。 常规的临床和实验室参数不足以检测正在进行的 疾病活动或对特定免疫调节剂的反应，强调了更好地描绘 潜在的免疫变化驱动了疾病活动的不可预测的爆发。缺乏知识 关于导致SLE病的精确免疫细胞和分子事件，构成了明显的障碍 为了开发准确的预后疾病生物标志物和选择性治疗剂。我们的长期 术语目标是鉴定可以提供治疗靶标的免疫细胞和分子介质 干涉。该建议的目的是阐明血清交流因子的机制 包括外泌体调节小儿SLE发病机理中的免疫细胞和细胞因子扰动。这 拟议研究的理由是，了解这些机制可以识别新型疾病 允许准确预后并提供选择性治疗靶标的生物标志物。 为了实现这一目标，HSIEH博士将使用高维质量细胞仪平台，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而不是荧光团作为结合的标签 抗体。在一项试点研究中，HSIEH博士建立了一个定量且可再现的质量细胞术测定法，该测定法 允许精确监测表型（表面标记）和功能性（细胞因子/趋化因子）免疫 在小儿SLE患者的外周血中可检测到的危险。这项研究的数据构成了基础 Hsieh博士的中心假设，即儿科SLE患者具有独特的多参数单核细胞 细胞因子特征，可驱动疾病活动，并由血清因子（包括外泌体）传播 通过激活JAK/STAT信号通路。在强大的初步数据的指导下，该假设将 通过追求三个特定目标来测试：1）确定细胞因子签名的关联（以及其他 免疫变化）通过整合质量细胞术免疫谱数据和临床 来自前瞻性纵向小儿SLE患者队列的参数； 2）阐明免疫调节 通过评估患病血清外泌体诱导的能力，SLE血清分离的外泌体的特性 细胞因子特征，并鉴定出对这种活性必不可少的外泌体蛋白质组学成分； 3） 通过确定鲁索利替尼（JAK1/2抑制剂）通过消除细胞因子特征表达的能力， 评估ruxolitinib对免疫的体内免疫抑制作用（包括靶向效应） 在小儿SLE发病机理中观察到的危险。 Hsieh博士是科罗拉多大学丹佛分校/科罗拉多州科罗拉多州科罗拉多州科罗拉多大学的助理教授 免疫学和微生物学和儿科部门。作为临床研究员，她在加里博士接受了培训 诺兰的实验室（斯坦福），被公认为在质量细胞术中的世界一流 分析，她获得了这项技术的专业知识。 Hsieh博士组建了一个指导团队 由具有基础科学免疫学研究专业知识的多学科科学家组成， 自身免疫性的免疫信号改变；临床研究设计和实施；外泌体 生物学;和生物统计学。她的主要导师约翰·坎比尔（John Cambier）博士和同事V.迈克尔·霍尔斯（V. Michael Holers）博士很好 建立的科学家合作着重于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业知识将为受训者提供丰富的经验 联合注册者迈克尔·格兰纳（Michael Graner）博士，国际外部生物学领域的国际知名专家， 顾问/合作者Jennifer Suep博士，儿科风湿病学家临床研究的重点是SLE和 生物统计学主席Debashis Ghosh博士，具有高通量数据分析的专业知识。共同 该团队将提供出色的培训环境，以填补她的工具箱中的关键空白和 知识库以增强她研究小儿SLE患者免疫危险的能力。 Hsieh博士有 制定了一项职业发展计划，将使她成功过渡到独立， 在：1）基础科学免疫学中纳入教学和指导培训； 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，拟议的方法是创新的，因为它偏离了现状 阐明SLE中炎症扰动的基本特定机制（例如，血清外泌体介导的 免疫调制）并通过表征这些机制，因为它们在多种免疫的背景下发生 单细胞水平 解决。拟议的研究很重要，因为它可能会确定候选疾病生物标志物 SLE中的预后和治疗目的 血清外泌体及其蛋白质组学成分。获得的知识将具有广泛的翻译 通过将这种系统免疫学方法应用于其他系统性风湿病的重要性 疾病。

项目成果

A great disturbance in the force: IL-2 receptor defects disrupt immune homeostasis.

• DOI: 10.1097/mop.0000000000001181
• 发表时间: 2022-12-01
• 期刊: CURRENT OPINION IN PEDIATRICS
• 影响因子: 3.6
• 作者: Hernandez, Joseph D.;Hsieh, Elena W. Y.
• 通讯作者: Hsieh, Elena W. Y.

Expansion of extrafollicular B and T cell subsets in childhood-onset systemic lupus erythematosus.

儿童期发病的系统性红斑狼疮中叶外 B 细胞和 T 细胞亚群的扩增。

• DOI: 10.3389/fimmu.2023.1208282
• 发表时间: 2023
• 期刊: Frontiers in immunology
• 影响因子: 7.3

Compositional Data Analysis using Kernels in mass cytometry data.

• DOI: 10.1093/bioadv/vbac003
• 发表时间: 2022
• 期刊: Bioinformatics advances
• 作者: Rudra P;Baxter R;Hsieh EWY;Ghosh D
• 通讯作者: Ghosh D

Peripheral immunophenotyping of AITD subjects reveals alterations in immune cells in pediatric vs adult-onset AITD.

• DOI: 10.1016/j.isci.2021.103626
• 发表时间: 2022-01-21
• 期刊: iScience
• 影响因子: 5.8
• 作者: Stensland ZC;Coleman BM;Rihanek M;Baxter RM;Gottlieb PA;Hsieh EWY;Sarapura VD;Simmons KM;Cambier JC;Smith MJ
• 通讯作者: Smith MJ

Proteomic profiling identifies key differences between inter-stage infants with single ventricle heart disease and healthy controls.

• DOI: 10.1016/j.trsl.2020.10.001
• 发表时间: 2021-03
• 期刊: Translational research : the journal of laboratory and clinical medicine
• 作者: Frank BS;Khailova L;Silveira L;Mitchell MB;Morgan GJ;Hsieh EWY;DiMaria MV;Twite M;Klawitter J;Davidson JA
• 通讯作者: Davidson JA