Mapping spatiotemporal dynamics during enterovirus infection across cells and tissues

项目摘要 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 皮肤粘膜损害，或手足口病。在某些情况下，感染就是生命- 具有威胁性。临床表现与其性向有关。比如一些混合动力车， 包括EV-A71和EV-D68在内的病毒与严重的神经症状有关，原因是 中枢神经组织的侵袭和损害。其他病毒，如柯萨奇病毒B3型，与呼吸道有关 症状和心肌病。这项建议旨在采取综合的方法来理解 这些组织和这些组织内的细胞如何对CVB3和新兴的 病原体EV-D68和EV-A71。我们将确定混合动力汽车的时间和空间动力学 利用基因组学的最新进展感染：(I)我们将分析单细胞转录本以 量化跨细胞和组织的病毒复制水平和宿主对感染的反应 感染的过程。(2)鉴于宿主内部适应似乎对感染很重要， 同时，我们将使用一种新的超深层技术绘制正在复制的病毒的突变谱 排序方法。我们将使用新的创新技术，如超深病毒 人口测序、深度学习和单细胞分析，以增加我们的基本理解 肠道病毒A、B和D的致病机制。最后，鉴于先天免疫是主要的 组织嗜性的决定因素，我们将使用具有特定I型干扰素亚型缺失的小鼠来 确定干扰素多样性在控制HEV感染中的意义。这些数据将 使我们能够确定HEV感染的细胞类型，即宿主对其的反应 在每个细胞和组织中，以及在不同组织中出现的病毒突变体。理解 发病机制是发展有效和广泛作用的对策的关键 并为有效和广谱疫苗和抗病毒药物的开发提供信息 化合物。

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.