Manipulation of Host Signature Expression Following Respiratory Virus Infection
EMSL Project ID
60513
Abstract
Pathogen invasion of host cells causes a myriad of functional changes including alterations of chromatin accessibility often limiting defense responses, shunting of cellular resources to centers of viral replication, and rearrangement of intracellular membranes to facilitate genome reproduction and progeny release. Systems biology approaches provide global snapshots of pathogen induced changes following infection and provide a variety of tools to begin to define how cellular homeostasis is disrupted, but improvements on these tools are required to determine how cellular functions are altered post infection. Primary human lung cells, grown in isolation or allowed to differentiate and grow at air liquid interface, provide an infection model that recapitulates the human conducting airway epithelium and an excellent ex vivo system to study human lung responses to infection. Protein-protein interactions and modifications of protein structure/function by the addition or removal of a metabolite regulate the activity of many host processes, however, these dynamic interactions are not typically captured by traditional proteomics approaches. The proposed studies are designed to use ex vivo models of the human bronchial airway, cutting edge, complementary and integrative proteomics techniques, scalable sample collection platforms, and activity-based probes to characterize how human respiratory viruses modify host responses in infected primary human lung cultures over time. Parallel studies will adapt a novel high throughput sample collection platform from prokaryotic to eukaryotic based growth conditions and examine how infection alters the exometabolome and secretome over time in a system that is designed for high-throughput automation. Findings from the first two studies will identify host enzymatic targets for evaluating activity-based probes to further characterize how infection alters normal host function. Experimental design and data analysis will be performed in collaboration with statisticians and computational modeling groups to ensure our results are significant and useful for model building and testing. In addition to increasing our knowledge base on how pathogens usurp the host environment during infection, this proposal will establish workflows for integrated and untargeted proteomics approaches to explore at a global level the cellular interactome and evaluate a novel high throughput sample capture platform for use with infected mammalian cells with the potential for downstream automated processes.
Project Details
Start Date
2022-06-14
End Date
N/A
Status
Active
Released Data Link
Team
Principal Investigator
Co-Investigator(s)
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