Project 1: Understanding the molecular pathways in SLE pathogenesis

项目主任/首席调查员(最后、第一、中间)：PD：帕斯夸尔，V./PI：项目1，帕斯夸尔，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 和发展针对核抗原的自身抗体。系统性红斑狼疮在临床上是异质性的，在分子水平上。 多种多样。这种异质性可能导致临床试验失败的高发生率，突显出 需要生物标记物根据疾病的个体致病驱动因素对患者进行分层。为了试图 了解系统性红斑狼疮的复杂性，我们建立了一个儿科队列，并在过去十年中对其进行了跟踪 使用经过验证的临床疾病活动性(DA)测量以及在耀斑期间的血液基因表达谱 和减刑。我们的研究证实了干扰素、中性粒细胞/髓系细胞和浆母细胞基因的流行 签名及其在队列层面上与发展援助的相关性。个性化免疫监测显示， 然而，在个体患者中，这些主要特征与DA的相关性存在显著的异质性 水平。我们假设解码这些签名的细胞和/或分子成分- 确定的患者群体将使生物标记物和分层计算工具的开发成为可能， 这将使合理的临床试验设计成为可能。 为了实现这一目标，我们提出了两个目标：1)确定三大 单细胞水平的系统性红斑狼疮血液特征。我们将检查产生这些签名的细胞 在群体和单细胞水平上使用高清晰度免疫表型和转录图谱；2) 确定分子DA标记是否与胞浆和/或内体核酸变化相关 (NA)体外患者血细胞和体外检测中的传感通路。在这里，我们首先提出申请 一种灵敏而可靠的方法来定量cGAS的内源活性，cGAS是一种通用的胞质DNA传感器，在 在病情发作和缓解期间患者的PBMCs。第二，我们将测试患者细胞对 相关的胞体和胞浆核酸配体在体外使用多维读数。通过 为了实现我们的目标，我们将一)揭示系统性红斑狼疮分子签名的来源；二)了解 血液中SLE髓系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感受器有助于免疫激活，导致系统性红斑狼疮发作。了解系统性红斑狼疮 异质性和在临床环境中评估它的工具的开发最终将开辟新的道路 个性化的治疗方法。

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.