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

> **NIH NIH F31** · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · 2021 · $41,081

## Abstract

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.

## Key facts

- **NIH application ID:** 10174643
- **Project number:** 5F31AI150007-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
- **Principal Investigator:** Sara Sunshine
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $41,081
- **Award type:** 5
- **Project period:** 2020-02-01 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10174643

## Citation

> US National Institutes of Health, RePORTER application 10174643, Identification and Characterization of the Antiviral Response to RSV Infection: A Single Cell Approach (5F31AI150007-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10174643. Licensed CC0.

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