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: 9356314
• 负责人: Maria Virginia Pascual
• 金额: $ 68.37万
• 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 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9356314
• 关键词: Affect; Antigen-Antibody Complex; Autoantibodies; Autoimmune Diseases; B-Lymphocytes; Biological Assay; Biological Markers; Blood; Blood Cells; Cells; Childhood; 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; Failure; Flare; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; Genomics; Goals; Hereditary Disease; Heterogeneity; Human; Immune system; Immunity; Immunophenotyping; In Vitro; Individual; Inflammation; Inflammatory; Interferon Type I; Interferons; Ligands; Lupus; Measures; Molecular; Molecular Profiling; Myelogenous; Myeloid Cells; Natural Immunity; Nuclear Antigens; Nucleic Acid Binding; Nucleic Acids; Pathogenesis; Pathogenicity; Pathway interactions; Patients; 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 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.

项目负责人/主要研究者（最后一名、第一名、中间名）：PD：Pascual，V. / PI：项目1 Pascual，V. 项目摘要 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 以及产生抗核抗原的自身抗体。SLE在临床上是异质性的， 多样化。这种异质性可能导致临床试验失败的高发生率，强调了 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。以试图 由于了解SLE的复杂性，我们建立了一个儿科队列，并在过去十年中对其进行了随访 使用经验证的临床疾病活动性（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流行 签名及其在队列水平上与DA的相关性。个性化免疫监测显示， 然而，在个体患者中，这些主要特征如何与DA相关的显著异质性 水平我们假设，解码这些签名的细胞和/或分子成分在良好的- 确定的患者组将能够开发用于分层的生物标志物和计算工具， 这将使临床试验设计合理。 为了实现这一目标，我们提出了两个目标：1）确定三个主要的起源和组成， 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晰度免疫表型分析和转录谱分析; 2） 确定分子DA标记物是否与细胞质和/或内体核酸的改变相关 (NA)在离体患者血细胞和体外测定中的感测途径。在此，我们首先提出， 一种灵敏和稳健的测定方法，用于定量cGAS（一种通用胞质DNA传感器）的内源性活性， 发作和缓解期间患者的PBMC。其次，我们将测试患者细胞对 使用多维读数在体外测定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传感器有助于导致SLE发作的免疫激活。了解SLE 异质性和开发工具来评估它在临床环境中将最终开辟新的途径， 个性化的治疗方法。