Endothelial Cell and Cardiomyocyte Dysfunction in Children with Kawasaki disease-like SARS-CoV-2 Induced Immune Activation

项目摘要 新冠肺炎大流行后，发热伴严重的全身炎症和休克，称为儿科 炎症性多系统综合征(PIMS)已成为儿童面临的新威胁。PIMS最初是 报告在西欧，美国的病例数量正在迅速增加。PIMS的一个标志是 心力衰竭导致休克，没有明显的肺部疾病。临床表现 这些患者与川崎病(KD)有许多相似之处，KD是获得性心脏病的最常见原因 儿童疾病，其本身可表现为分布性休克，需要肌力和血管活性支持 重症监护室。几名PIMS患者要么SARS-CoV-2聚合酶链式反应阳性，要么已经发展成 抗SARS-CoV-2抗体，提示PIMS是对既往暴露的免疫介导反应 对病毒来说。奇怪的是，在患者被诊断为PIMS的同时， 在这些相同的地区，典型的KD病例急剧增加。PIMS的出现是如此之新，因此 迅速发展，几乎没有与这些疾病相关的出版物、治疗指南或临床试验 受影响严重的儿科患者。通过我们参与KidCare的30个儿科中心网络，我们的 由PCORI资助的耐药KD的比较有效性试验，我们正在收集患者数据和 临床样本支持这里提出的工作。这份行政副刊的目的是分析 人口学、临床和实验室数据，结合使用患者细胞和血清的分析，这 将使我们能够研究SARS-CoV-2感染、典型KD和PIMS之间的关系并建立模型 针对PIMS的不同治疗策略。为实现这一目标提出了三个具体目标。特定的 目的1将描述和比较PIMS和典型KD的临床特征。人口学和临床数据以及 将比较这两种疾病中性粒细胞对静脉注射免疫球蛋白(IVIG)的反应。 急性和亚急性PIMS和典型KD患者血浆中的细胞因子和炎症标志物也将 被比较。特异性目标2将阐明PIMS的分子特征，并与典型的KD进行比较。我们会 使用rna-seq、elisa和Western印迹分析来描述与炎症相关的分子水平的变化。 (如TIFA、NFKβ、NLRP3-炎症体、IL-1、IL-6、肿瘤坏死因子α)和心血管健康(KLF4、MIR-483、ACE2) 在内皮细胞和心肌细胞中。具体目标3将通过以下方式测试PIMS的药物治疗效果 静脉注射免疫球蛋白、类固醇和阿纳金纳对炎症通路影响的体外比较 急性PIMS患者血清治疗后内皮细胞和心肌细胞中的心血管生物标志物 患者在治疗前。在本多PI补充资料中，调查小组的协同专业知识提供 了解PIMS的临床特征、分子基础和药物治疗效果的难得机会 与KD相比。

Project Summary In the wake of COVID-19 pandemic, fever with severe systemic inflammation and shock, known as Pediatric Inflammatory Multisystem Syndrome (PIMS), has evolved as a new threat to children. PIMS was originally reported in Western Europe and the number of cases is rapidly increasing in the U.S. A hallmark of PIMS has been heart failure leading to shock and the absence of significant pulmonary disease. The clinical presentation in these patients shares many features with Kawasaki disease (KD), the most common cause of acquired heart disease in children, which itself can present with distributive shock requiring inotropic and vasoactive support in the intensive care unit. Several PIMS patients are either SARS-CoV-2 PCR positive or have developed antibodies against SARS-CoV-2, suggesting that PIMS is an immune-mediated reaction to antecedent exposure to the virus. Curiously, at the same time that patients are being diagnosed with PIMS, the numbers of children with typical KD has increased dramatically in these same regions. The emergence of PIMS is so new and so rapidly evolving that there are literally no publications, treatment guidelines or clinical trials related to these seriously affected pediatric patients. Through our network of 30 pediatric centers participating in KIDCARE, our comparative effectiveness trial for treatment-resistant KD funded by PCORI, we are collecting patient data and clinical samples to support the work proposed here. The goal of this administrative supplement is to analyze demographic, clinical and laboratory data in conjunction with assays using patient cells and sera, which will allow us to study the relationships among SARS-CoV-2 infection, typical KD, and PIMS and to model different therapeutic strategies against PIMS. Three specific aims are proposed to achieve this goal. Specific Aim 1 will profile and compare clinical features of PIMS and typical KD. Demographic and clinical data and the neutrophil response to intravenous immunoglobulin (IVIG) will be compared between these two illnesses. Cytokine profiles and inflammatory markers in plasma from acute and subacute PIMS and typical KD will also be compared. Specific Aim 2 will elucidate molecular features of PIMS and compare with typical KD. We will use RNA-seq, ELISA, and Western blot analyses to profile changes in levels of molecules related to inflammation (e.g., TIFA, NFkβ, NLRP3-inflammasome, IL-1, IL-6, TNFα) and cardiovascular health (KLF4, miR-483, ACE2) in endothelial cells and cardiomyocytes. Specific Aim 3 will test the efficacy of drug therapy for PIMS by comparing the in vitro effects of intravenous immunoglobulin, steroids, and anakinra on inflammatory pathways and cardiovascular biomarkers in endothelial cells and cardiomyocytes treated with sera from acute PIMS patients prior to therapy. The synergistic expertise of the investigative teams in this multi-PI supplement provides a unique opportunity to understand the clinical features, molecular basis, and efficacy of drug treatment of PIMS as compared to KD.