A spatially resolved molecular atlas of acute viral myocarditis at single-cell resolution

单细胞分辨率的急性病毒性心肌炎的空间解析分子图谱

基本信息

批准号：
10681925
负责人：
Iwijn De Vlaminck
金额：
$ 77.68万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-15 至 2028-02-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10681925
关键词：
Acute Address Area Arrhythmia Atlases Bioinformatics Biological Biological Assay Biological Markers Biopsy Blood Bystander Effect Cardiac Cardiac Myocytes Cell Communication Cell Death Cells Child Complex Cytotoxic T-Lymphocytes Data Diagnosis Diagnostic Diagnostic Sensitivity Dilated Cardiomyopathy Disease Disease Outcome Endothelial Cells Endothelium Environment Genetic Transcription Heart Heterogeneity Host Defense Immune In Situ Infant Infection Infiltration Inflammatory Injury Innate Immune Response Integration Host Factors Interferons Knockout Mice Knowledge Mammalian Orthoreovirus Maps Measurement Measures Mediating Medical Methodology Modeling Molecular Monitor Mus Myocarditis Nature Neonatal Onset of illness Outcome Pathogenesis Pathologic Processes Plasma Play Procedures Process RNA Reovirus Reovirus Infections Resolution Role Sampling Severity of illness Sudden Death Tail Techniques Technology Testing Time Tissues Viral Viral Pathogenesis Virus Virus Diseases Work blood-based biomarker cell injury cell type cross-species transmission data integration diagnosis standard diagnostic biomarker diagnostic strategy experimental study heart damage improved inhibitor innovation insight liquid biopsy minimally invasive mouse model mutant neonatal mice neonate noninvasive diagnosis novel novel diagnostics novel strategies recruit response sex single-cell RNA sequencing spatiotemporal success therapeutic target tool transcriptome sequencing transcriptomics viral RNA viral myocarditis virology

项目摘要

Viral myocarditis is a heterogeneous disease that is difficult to study and diagnose. Because of the heterogeneous nature of acute viral myocarditis and the difficulty and low diagnostic sensitivity of endomyocardial biopsies (considered the gold standard for diagnosis of viral myocarditis), there is limited knowledge of the molecular pathogenesis of this disease, particularly in infants and neonates where endomyocardial biopsies are less often performed. The specific cells within the heart that respond to viral infection, the nature of their responses, and the spatiotemporal distribution of such responses are not well known. A better understanding of the spatiotemporal response of the heart to viral infection at the cellular and molecular level will provide much needed insight into the pathological processes that drive the active inflammatory process that ensues following viral infection of the heart. The lack of understanding of the molecular pathogenesis of viral myocarditis is in part due to the lack of tools to investigate viral infection in complex native tissues at single cell-resolution. Here we will use innovative spatially resolved transcriptomics, single-cell RNA sequencing (scRNA-seq) tools and bioinformatics, in conjunction with classical virology techniques, and mouse models to study myocarditis in mammalian orthoreovirus (REOV). We have three aims: In Aim 1, we will study the viral and host factors that define the outcome of REOV infection of cardiac tissues. In Aim 2, we will determine the role of pyroptosis in REOV-induced myocarditis. In Aim 3, we will combine our high-resolution single-cell atlas of myocarditis with the principles of liquid biopsies based on cell-free RNA to develop highly specific blood biomarkers of viral myocarditis, thereby addressing an urgent and unmet medical need. We anticipate our studies will provide unprecedented insight into the pathobiology of viral myocarditis. Our experiments will clarify which cell types are infected in complex cardiac tissues and will reveal how infection success depends on both cell state and cellular environment. We will elucidate the role for endothelial cells in viral myocarditis and we will explore whether and how immune cell responses switch from host defense to host injury. We will explore the effects of infected cells on uninfected bystander cells in close physical proximity, and we will map the cellular interactions that mediate this bystander effect. We will also explore the spatial and cell type heterogeneity of innate immune responses within infected and uninfected cardiac tissues. Successful implementation of these studies will lead to new approaches and molecular tools to study viral myocarditis and other viral diseases and may identify novel diagnostic approaches and therapeutic targets for acute viral myocarditis.

病毒性心肌炎是一种难以研究和诊断的异质性疾病。因为急性病毒性心肌炎的异质性以及诊断难度和敏感性低心内膜心肌活检（被认为是诊断病毒性心肌炎的金标准），但效果有限了解这种疾病的分子发病机制，特别是婴儿和新生儿心内膜心肌活检较少进行。心脏内对病毒做出反应的特定细胞感染、其反应的性质以及此类反应的时空分布尚不清楚。从细胞和分子角度更好地了解心脏对病毒感染的时空反应水平将为驱动活跃炎症过程的病理过程提供急需的见解这是心脏病毒感染后发生的。对病毒性心肌炎的分子发病机制缺乏了解，部分原因是缺乏诊断病毒性心肌炎的工具。以单细胞分辨率研究复杂天然组织中的病毒感染。在这里我们将利用创新的空间解析转录组学、单细胞 RNA 测序 (scRNA-seq) 工具和生物信息学，并结合经典病毒学技术和小鼠模型来研究哺乳动物正呼肠孤病毒（REOV）的心肌炎。我们有三个目标：在目标 1 中，我们将研究决定 REOV 感染结果的病毒和宿主因素心脏组织。在目标 2 中，我们将确定细胞焦亡在 REOV 诱导的心肌炎中的作用。在目标 3 中，我们将把我们的心肌炎高分辨率单细胞图谱与基于液体活检的原理相结合无细胞RNA来开发病毒性心肌炎的高度特异性血液生物标志物，从而解决一个紧迫的问题未满足的医疗需求。我们预计我们的研究将为病毒病理学提供前所未有的见解心肌炎。我们的实验将阐明哪些细胞类型在复杂的心脏组织中被感染，并将揭示感染成功与否取决于细胞状态和细胞环境。我们将阐明其作用病毒性心肌炎中的内皮细胞，我们将探讨免疫细胞反应是否以及如何从宿主对宿主伤害的防御。我们将近距离探索受感染细胞对未受感染旁观者细胞的影响物理上的接近，我们将绘制介导这种旁观者效应的细胞相互作用。我们也会探索感染者和未感染者先天免疫反应的空间和细胞类型异质性心脏组织。这些研究的成功实施将带来研究病毒的新方法和分子工具心肌炎和其他病毒性疾病，并可能确定新的诊断方法和治疗靶点急性病毒性心肌炎。