Project Details

Inhibition of the RalGAP protein complex as a novel approach to broadly block enteric infections

H. Pascaline Kohio, PhD

Summary

BACKGROUND: Rotavirus, enterohemorrhagic Escherichia coli, and Cryptosporidium parvum are the principal viral, bacterial, and parasitic pathogens that cause severe gastroenteritis and life-threatening diarrhea in infant and young children. Rotavirus vaccine efficacy is low in endemic countries and there are no effective therapeutics or vaccines against enterohemorrhagic Escherichia coli, and Cryptosporidium parvum. GAP: Currently, there is a lack of knowledge on the role of host factors that support enteric pathogens infection which will help design novel antiviral therapeutics. However, our previous CRISPR/Cas9 screens and published literature suggest that genetic deletion of the RalGAP complex suppresses rotavirus, enterohemorrhagic Escherichia coli, and Cryptosporidium parvum infections. HYPOTHESIS: Given the role of RalGAP in endosomal vesicular trafficking, we speculate that these pathogens commonly hijack the RalGAP complex for endosomal entry and toxin release into the host intestinal epithelial cells. We hypothesize that the inhibition of host RalGAP complex will block the replication and pathogenesis of rotavirus, Escherichia coli, and Cryptosporidium parvum. METHODS: RALGAPA1 gene will be genetically deleted via CRISPR/Cas9 in human intestinal cell cultures and neonatal mice; and rotavirus, enterohemorrhagic Escherichia coli, and Cryptosporidium parvum infections and diseases will be assessed in vivo and in vitro. We will screen for novel small molecules that can inhibit RALGAP activity. RESULTS: Pending. IMPACT: We expect our bench-to-bedside discovery pipeline to help identify the host cellular druggable targets commonly usurped by these enteric pathogens and develop a series of broadly anti-microbial molecules to treat enteric infections, thereby reducing diarrhea-associated morbidity and mortality. Website Link: https://sdinglab.wustl.edu/people-page-page/