Spatial-Temporal Dissection of Stratified Host Tissue Responses to Severe acute respiratory syndrome-related coronaviruses in situ to Understand Intra-host Pathogenesis

对严重急性呼吸综合征相关冠状病毒的分层宿主组织反应进行时空解剖，以了解宿主内发病机制

• 批准号: 10698159
• 负责人: Sizun Jiang
• 金额: $ 47.5万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-06 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698159
• 关键词: 2019-nCoV; ACE2; Address; Affect; Animal Model; Antibodies; Autopsy; Bar Codes; Biological; Biological Models; COVID-19; COVID-19 pandemic; Cancer Biology; Cells; Cessation of life; Clinical; Communicable Diseases; Computing Methodologies; Coronavirus; Coupled; Data Analyses; Developmental Biology; Disease; Disease model; Dissection; Engineering; Epitopes; Eukaryota; Host Defense; Human; Immune; Immune response; Immune system; In Situ; Individual; Infection; Infectious Agent; Integration Host Factors; Investigation; K-18 conjugate; Methodology; Minor; Modeling; Molecular Virology; Mus; Mutation; Nucleic Acids; Pathogenesis; Proteins; RNA Viruses; Research Proposals; Rodent; SARS-CoV-2 variant; Sampling; Severe Acute Respiratory Syndrome; System; Technology; Therapeutic; Tissue imaging; Tissues; Tropism; Vaccines; Variant; Viral; Viral Pathogenesis; Virus; Virus Diseases; Work; biological systems; combinatorial; data integration; design; imaging platform; in situ imaging; in vivo; innovation; insight; invention; mouse model; neutralizing antibody; new technology; next generation; nonhuman primate; novel; pathogen; pathogenic virus; prevent; protein aminoacid sequence; response; reverse genetics; technology platform; theories; tissue tropism; vaccine efficacy; variants of concern

PROJECT SUMMARY/ABSTRACT Distinctive host-pathogen interactions and adaptations are the cornerstones of co-evolution between eukaryotes and their viral pathogens. Given the rapid replication and mutation rates of RNA viruses within their host, understanding these intra-host interactions in their native tissue context is central to developing next-generation anti-viral and vaccines. The ongoing COVID-19 pandemic continues to thwart eradication due to SARS-CoV-2 Variants of Concern (VoC), where even minor changes to the spike protein can affect cellular entry, antibody neutralization, vaccine efficacy and immune responses, thus leading to immune escape. Why do these variants, along with other Severe acute respiratory syndrome–related coronaviruses (SARSr-CoVs), differ in their host pathogenesis, and how do they achieve that? Are there differential host factors or responses that influence tissue-specific tropism? These questions need to be answered with controlled experimental approaches to dissect and deconvolute the coevolutionary viral-host interactions in situ. This proposal seeks to combine powerful reverse genetics systems, next-generation tissue imaging platforms and robust animal models towards the systematic determination of host immune responses, virus evasion strategies and both inter- and intra-host viral dynamics. We propose to use these powerful methodologies to first determine host tissue responses to individual VoCs within human COVID-19 autopsies and non-human primate necropsies. Next, we seek to combine spatial-lineage tracing with tagged SARS-CoV-2 VoCs and other SARSr-CoVs that use ACE2 for entry in the humanized ACE2-K18 mouse model. To recapitulate intra-host viral variation in vivo, we will engineer these viruses to include a short sequence of peptides encoding unique barcodes. Each set of barcodes corresponds to a specific virus strain or variant. Infection of humanized ACE2 mice with a pool of these barcoded viruses, coupled with antibody or hybridization-based spatial readouts and single-cell characterization of host immune responses, will enable a methodical approach towards the systems-level investigation of intra-host viral variation and competition dynamics. The spatial framework and conceptual advances resulting from this work are applicable to a broad myriad of other biological systems and diseases, thus paving the way to better- designed therapeutics and rapid responses to understand, control and eventually eradicate new biological threats.

项目概要/摘要 独特的宿主-病原体相互作用和适应是真核生物之间共同进化的基石 及其病毒病原体 鉴于 RNA 病毒在宿主体内的快速复制和突变率， 了解宿主在其天然组织环境中的这些相互作用对于开发下一代至关重要 由于 SARS-CoV-2，持续的 COVID-19 大流行继续阻碍根除。 关注变体 (VoC)，即使刺突蛋白发生微小变化也会影响细胞进入、抗体 中和作用、疫苗功效和免疫反应，从而导致这些变异， 与其他严重急性呼吸综合征相关冠状病毒（SARSr-CoV）一样，它们的不同之处在于 宿主发病机制，以及它们如何实现这一目标？是否存在不同的宿主因素或反应？ 影响组织特异性向性？这些问题需要通过受控实验来回答 该提案旨在剖析和解算原位共同进化病毒-宿主相互作用的方法。 结合强大的反向遗传学系统、下一代组织成像平台和强大的动物模型 宿主免疫反应、病毒逃避策略以及内部和外部的系统确定 我们建议使用这些强大的方法首先确定宿主组织。 接下来，我们对人类 COVID-19 尸检和非人类灵长类动物尸检中个体 VoC 的反应。 寻求将空间谱系追踪与标记的 SARS-CoV-2 VoC 和其他使用 ACE2 的 SARSr-CoV 结合起来 为了进入人源化 ACE2-K18 小鼠模型，我们将在体内重现宿主内的病毒变异。 对这些病毒进行改造，使其包含编码每组条形码的短肽序列。 用这些条形码库感染人源化 ACE2 小鼠 病毒，结合抗体或基于杂交的空间读数和宿主的单细胞表征 免疫反应，将为宿主内病毒的系统级研究提供系统方法 这项工作带来的空间框架和概念进步。 适用于无数其他生物系统和疾病，从而为更好地- 设计治疗方法和快速反应来理解、控制并最终根除新的生物 威胁。

项目成果

Morphologic and molecular analysis of liver injury after SARS-CoV-2 vaccination reveals distinct characteristics.

SARS-CoV-2 疫苗接种后肝损伤的形态学和分子分析揭示了独特的特征。

• 发表时间: 2023-09
• 影响因子: 25.7
• 作者: Uzun, Sarp;Zinner, Carl P;Beenen, Amke C;Alborelli, Ilaria;Bartoszek, Ewelina M;Yeung, Jason;Calgua, Byron;Reinscheid, Matthias;Bronsert, Peter;Stalder, Anna K;Haslbauer, Jasmin D;Vosbeck, Juerg;Mazzucchelli, Luca;Hoffmann, Tobias;Terraccian
• 通讯作者: Terraccian