Project 1: Understanding the molecular pathways in SLE pathogenesis

项目 1：了解 SLE 发病机制的分子途径

• 批准号: 9891960
• 负责人: Maria Virginia Pascual
• 金额: $ 61.06万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9891960
• 关键词: Affect; Antigen-Antibody Complex; Autoantibodies; Autoimmune Diseases; B-Lymphocytes; Biological Assay; Biological Markers; Blood; Blood Cells; Cells; Clinical; Clinical Trials; Clinical Trials Design; Complement; DNA; Data; Defect; Dendritic Cells; Dendritic cell activation; Development; Diagnostic; Differentiation Antigens; Disease; Disease Marker; Disease remission; Disease susceptibility; Expression Profiling; Failure; Flare; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; Genomics; Goals; Heterogeneity; Human; Immune system; Immunity; Immunologic Monitoring; Immunophenotyping; In Vitro; Individual; Inflammation; Inflammatory; Interferon Type I; Interferons; Ligands; Lupus; Measures; Molecular; Molecular Disease; Molecular Profiling; Myelogenous; Myeloid Cells; Natural Immunity; Nuclear Antigens; Nucleic Acid Binding; Nucleic Acids; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pediatric cohort; Peripheral Blood Mononuclear Cell; Plasma Cells; Plasmablast; Population; Prevalence; Principal Investigator; Production; Research; Resources; Sampling; Source; Stratification; Systemic Lupus Erythematosus; Testing; Therapeutic Intervention; Transcript; Transcriptional Activation; Virginia; activity marker; adaptive immunity; autoreactive B cell; biomarker development; cell type; cohort; complement deficiency; computerized tools; cytokine; genetic signature; genetic variant; genome wide association study; immune activation; in vitro Assay; individual patient; molecular marker; molecular subtypes; neutrophil; overexpression; patient response; patient stratification; personalized therapeutic; programs; response; sensor; tool

Program Director/Principal Investigator (Last, First, Middle): PD: Pascual, V. / PI: Project 1 Pascual, V. Project Summary Systemic Lupus Erythematosus (SLE) is an autoimmune disease characterized by widespread inflammation and development of autoantibodies against nuclear antigens. SLE is clinically heterogeneous and molecularly diverse. This heterogeneity might contribute to the high occurrence of clinical trial failures, underscoring the need for biomarkers to stratify patients according to individual pathogenic drivers of disease. In an attempt to understand the complexity of SLE, we established a pediatric cohort and have followed it for the past decade using validated clinical disease activity (DA) measures as well as blood gene expression profiles during flares and remissions. Our studies confirm the prevalence of IFN, neutrophil/myeloid and plasmablast gene signatures and their correlation with DA at the cohort level. Personalized immunomonitoring revealed, however, significant heterogeneity in how these major signatures correlate with DA at the individual patient level. We hypothesize that decoding the cellular and/or molecular components of these signatures in well- defined groups of patients will enable development of biomarkers and computational tools for stratification, which will enable rational clinical trial design. Towards this goal, we are proposing two aims: 1) to establish the origin and composition of three major SLE blood signatures at the single cell level. We will examine the cells that give rise to these signatures using high definition immunophenotyping and transcriptional profiling at the population and single cell levels; 2) to determine if molecular DA markers correlate with altered cytosolic and/or endosomal nucleic acid (NA) sensing pathways in ex vivo patient blood cells and in vitro assays. Here, we first propose to apply a sensitive and robust assay to quantify the endogenous activity of cGAS, a universal cytosolic DNA sensor, in PBMCs from patients during flares and remissions. Second, we will test the response of patient cells to relevant endosomal and cytosolic nucleic acid ligands in vitro using multi-dimensional readouts. Through the implementation of our aims, we will i) reveal the source of SLE molecular signatures; ii) understand the extent of heterogeneity of blood SLE myeloid cells and plasma cells; iii) determine which cell subsets/molecular pathways and/or NA sensors contribute to immune activation leading to SLE flares. Understanding SLE heterogeneity and developing tools to assess it in the clinical setting will ultimately open new paths towards personalized therapeutic approaches.

项目总监/首席研究员（最后、第一、中间）： PD：Pascual, V. / PI：项目 1 Pascual, V. 项目概要 系统性红斑狼疮 (SLE) 是一种以广泛炎症为特征的自身免疫性疾病 和针对核抗原的自身抗体的开发。 SLE 具有临床异质性和分子水平 各种各样的。这种异质性可能导致临床试验失败的高发生率，强调 需要生物标志物根据疾病的个体致病驱动因素对患者进行分层。试图 了解 SLE 的复杂性后，我们建立了一个儿科队列并在过去十年中对其进行了跟踪 使用经过验证的临床疾病活动 (DA) 测量以及发作期间的血液基因表达谱 和缓解。我们的研究证实了 IFN、中性粒细胞/髓细胞和浆母细胞基因的患病率 特征及其与队列水平 DA 的相关性。个性化免疫监测显示， 然而，这些主要特征与个体患者的 DA 的相关性存在显着的异质性 等级。我们假设解码这些特征的细胞和/或分子成分 定义的患者群体将能够开发用于分层的生物标志物和计算工具， 这将使合理的临床试验设计成为可能。 为实现这一目标，我们提出两个目标：1）确定三大领域的起源和构成。 单细胞水平的 SLE 血液特征。我们将检查产生这些特征的细胞 在群体和单细胞水平上使用高清免疫表型分析和转录分析； 2） 确定分子 DA 标记是否与改变的胞质和/或内体核酸相关 (NA) 离体患者血细胞和体外测定中的传感途径。在这里，我们首先建议申请 一种灵敏而稳健的测定法，用于量化 cGAS（一种通用胞质 DNA 传感器）的内源活性 发作和缓解期间患者的 PBMC。其次，我们将测试患者细胞的反应 使用多维读数体外相关的内体和胞质核酸配体。通过 为了实现我们的目标，我们将 i) 揭示 SLE 分子特征的来源； ii) 了解程度 血液 SLE 骨髓细胞和浆细胞的异质性； iii) 确定哪些细胞亚群/分子 通路和/或 NA 传感器有助于免疫激活，导致 SLE 发作。了解系统性红斑狼疮 异质性和开发在临床环境中评估异质性的工具最终将开辟新的道路 个性化的治疗方法。