Mapping spatiotemporal dynamics during enterovirus infection across cells and tissues

绘制肠道病毒跨细胞和组织感染过程中的时空动态

• 批准号: 10875953
• 负责人: Raul Andino
• 金额: $ 9.83万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-10 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10875953
• 关键词: Acute; Adolescent; Affect; Alleles; Animal Model; Antiviral Agents; Antiviral Response; Atlases; Attenuated; Biological; Brain Stem; CRISPR/Cas technology; Cardiomyopathies; Cells; Clinical; Coxsackie B Viruses; Data; Data Set; Development; Diabetes Mellitus; Disease; Encephalitis; Enterovirus; Enterovirus 68; Enterovirus 71; Enterovirus Infections; Evolution; Family; Genes; Genetic Determinism; Genetic Structures; Genetic Transcription; Genomics; Goals; Hand, Foot and Mouth Disease; Heart Diseases; Hospitalization; Human; Human poliovirus; Immune response; Individual; Infant; Infection; Innate Immune Response; Interferon Type I; Interferon alpha; Interferon-beta; Interferons; Intervention; Invaded; Knowledge; Lesion; Life; Link; Maps; Measures; Medical; Meningitis; Methods; Mus; Mutation; Myocarditis; Natural Immunity; Nerve Tissue; Neurologic Symptoms; Pancreatitis; Paralysed; Pathogenesis; Pathogenicity; Pathway interactions; Pericarditis; Pharmaceutical Preparations; Phylogenetic Analysis; Population; Population Dynamics; Population Genetics; Process; Respiratory Signs and Symptoms; Reverse engineering; Role; Severities; Shapes; Signal Transduction; Source; Specificity; Structure; Testing; Tissue-Specific Gene Expression; Tissues; Tropism; United States; Vaccines; Viral; Virulence; Virus; Virus Diseases; Virus Replication; cell type; deep learning; deep sequencing; emerging pathogen; experimental study; in vivo; innovative technologies; insight; mathematical model; mouse model; mutant; neonatal sepsis; novel; pressure; prevent; respiratory; response; single cell analysis; single-cell RNA sequencing; spatiotemporal; tissue tropism; transcriptome; transcriptomics; vaccine development; virus genetics; virus host interaction

Project Summary Human enterovirus (HEV) infections primarily affect infant and adolescent populations, causing a wide range of clinical manifestations that commonly include respiratory illness and mucocutaneous lesions, or hand, foot and mouth disease. In some cases, the infection is life- threatening. The clinical manifestations are a function of their tropism. For example, some HEVs, including EV-A71 and EV-D68, are associated with serious neurological symptoms due to invasion and damage of central nervous tissues. Others, such CVB3, are implicated in respiratory symptoms and cardiomyopathy. This proposal aims to take an integrative approach to understand how tissues, and cells within these tissues, respond to infection by CVB3 and the emerging pathogens EV-D68 and EV-A71. We will determine the temporal and spatial dynamics of HEV infection using recent advances in genomics: (i) We will profile single-cell transcriptomes to quantify viral replication levels and the host response to infection across cells and tissues over the course of infection. (ii) Given that intra-host adaptation appears to be important in infection, in parallel, we will map the mutational spectrum of the replicating viruses using a novel ultra-deep sequencing approach. We will use new innovative technologies, such as ultra-deep virus population sequencing, deep learning and single-cell analysis to increase our basic understanding of the pathogenesis of enteroviruses A, B and D. Finally, given that innate immunity is a major determinant of tissue tropism, we will use mice with deletions of specific type-I IFN subtypes to determine the significance of interferon diversity in controlling HEV infections. These data will enable us to determine cell types that HEVs infect, the response that the host mounts against them in each cell and tissue, and the viral mutants that emerge in different tissues. Understanding pathogenesis is critically needed for developing effective and broadly-acting countermeasures and to inform the development of effective and broad-spectrum vaccines and antiviral compounds.

项目概要 人类肠道病毒 (HEV) 感染主要影响婴儿和青少年人群，导致 广泛的临床表现，通常包括呼吸道疾病和 皮肤粘膜病变，或手足口病。在某些情况下，感染是生命的 威胁。临床表现是其趋向性的函数。例如，一些混合动力汽车， 包括 EV-A71 和 EV-D68，由于以下原因与严重的神经系统症状相关 中枢神经组织的侵袭和损伤。其他的，如 CVB3，与呼吸道疾病有关。 症状和心肌病。该提案旨在采取综合方法来理解 组织以及这些组织内的细胞如何响应 CVB3 和新兴疾病的感染 病原体 EV-D68 和 EV-A71。我们将确定 HEV 的时间和空间动态 利用基因组学的最新进展来识别感染：（i）我们将分析单细胞转录组以 量化病毒复制水平以及宿主对细胞和组织感染的反应 感染过程。 (ii) 鉴于宿主内适应似乎在感染中很重要， 与此同时，我们将使用一种新型的超深度方法绘制复制病毒的突变谱图。 测序方法。我们将使用新的创新技术，例如超深度病毒 群体测序、深度学习和单细胞分析，以增加我们的基本了解 肠道病毒 A、B 和 D 的发病机制。最后，鉴于先天免疫是主要的 组织向性的决定因素，我们将使用具有特定 I 型 IFN 亚型缺失的小鼠来 确定干扰素多样性在控制 HEV 感染中的重要性。这些数据将 使我们能够确定 HEV 感染的细胞类型、宿主的反应 它们存在于每个细胞和组织中，以及出现在不同组织中的病毒突变体。理解 制定有效且广泛作用的对策迫切需要发病机制 并为开发有效的广谱疫苗和抗病毒药物提供信息 化合物。