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Identification and Characterization of the Antiviral Response to RSV Infection: A Single Cell Approach

RSV 感染抗病毒反应的鉴定和表征：单细胞方法

基本信息

批准号：
9907392
负责人：
Sara Sunshine
金额：
$ 4.06万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9907392
关键词：
A549 Antiviral Agents Antiviral Response Bioinformatics Biology Brefeldin A CRISPR screen Cell Separation Cells Clustered Regularly Interspaced Short Palindromic Repeats Complement Data Data Set Epithelial Epithelial Cells Epithelium Foundations Future Gene Activation Genes Genetic Transcription Heterogeneity Hour Human Human Genome ISG15 gene Immune response In Vitro Individual Infant Mortality Infection Inflammatory Response Integration Host Factors Interferons Knowledge Label Life Cycle Stages Microscopy Modeling Molecular Paracrine Communication Patients Phenotype Play Poly A Prevalence Process Production Protein Secretion Publishing Research Respiration Respiratory Syncytial Virus Infections Respiratory System Respiratory syncytial virus Role Scientist Structure of respiratory epithelium Techniques Testing Therapeutic Time Transcript Vaccines Viral Viral Load result Virion Virus Virus Diseases Virus Replication base cytokine experimental study genetic signature immunoregulation in vivo inhibitor/antagonist insight mutant novel therapeutics overexpression pathogen prevent programs receptor respiratory response single cell sequencing skills targeted treatment therapeutic development time use transcriptome

项目摘要

Project Summary/Abstract Respiratory Syncytial Virus (RSV) is a ubiquitous respiratory illness, and the leading viral cause of infant mortality worldwide. Despite its prevalence, there remains no effective vaccine, nor an efficacious RSV-specific therapeutic for infected patients. It is critical that we understand the fundamental biology underlying RSV infection and the subsequent host response to further our understanding of this pathogen, and lay the foundation for novel therapeutic strategies. The temporal dynamics of RSV infection have been characterized in bulk studies in vitro, in vivo, and ex vivo, however, these bulk studies likely underestimate the complexity and heterogeneity of the host respiratory epithelial response to infection. I have performed preliminary experiments to investigate the single cell response of respiratory epithelial cells (A549 cells) to RSV infection and my data revealed viral burden dependent and distinct gene networks activated over a twelve hour course of RSV infection. My data suggests a model in which infection may be considered a dynamic process with a spectrum of transcriptional states indicative of infection state. Indeed, I find expression of interferon stimulated genes (ISGs) is stronger in cells with undetectable RSV transcripts, compared to nearby infected cells. This is indicative of a compromised innate defense state within infected cells, but sensed by the surrounding cells. Given my preliminary studies, I propose a research strategy that seeks to test my hypotheses that the proinflammatory response during infection is largely driven by nearby uninfected cells, and that distinct proinflammatory gene signatures will correlate with different stages of the adjacent viral life cycle. In Aim 1, I will characterize the viral and host single cell temporal transcriptional dynamics of RSV infection in both infected and uninfected bystander cells, and produce single cell trajectories modeling RSV infection. Additionally, I will determine the contribution of paracrine signaling to the proinflammatory response initiated after RSV infection, by blocking protein secretion with Brefeldin A, and investigating the transcriptional response of uninfected bystander cells. In Aim 2, I will complement this study by performing an ISG-wide CRISPRa activation screen to identify host factors that influence both RSV replication and virion production. The completion of this study, which leverages single cell sequencing, bioinformatics, and CRISPR screening techniques, will provide pivotal insights into our understanding of the antiviral response to RSV infection.

项目总结/文摘