Development of High Throughput Screening and High Content Platforms for La Crosse Encephalitis Virus

Authors

• KH Berry
• RV Remigio
• MCR Alvanja
• G Andreotti
• P Albert
• WJ Lee
• DP Sandler
• JN Hofmann
• LE Beane Freeman

Abstract

The recent global pandemic has highlighted the pressing need for antiviral drug discovery tools in the effort to prevent future burdens on healthcare infrastructure and human health. Multiple RNA virus families have been identified as those presenting pandemic potential, including members of the order Bunyavirales. This negative-sense RNA virus order contains major human pathogens such Rift Valley Fever Virus, California-group Encephalitis viruses, and rodent borne Hantaviruses. Of these viruses, La Crosse Encephalitis virus (LACV) is known to cause pediatric encephalitis with typically only palliative care options available as therapeutic treatment, emphasizing the need for relevant model systems and antiviral therapeutics. Previous efforts have established a high-throughput screening (HTS) platform for anti-viral compounds using a cytoprotective assay. Here, we share work expanding this model and additionally developing a 1536-well, high content screening (HCS) platform based on direct detection of LACV glycoprotein in infected cell cultures for anti-viral screening. We observed a differential effect of several known anti-LACV compounds in indirect (e.g. cytoprotective assays) vs direct (e.g. IFA-based assay). We further examined the relationship between cytoprotective and direct viral antigen-based assay readouts by comparing the efficacy of established anti-virals and then directly compared the two assays by screening a library of 752 anti-infective compounds with both the HTS and HCS platforms, and report that a combination of these two assays provide orthogonal coverage to best determine lead anti-viral compounds.

Scientific Focus Area: Virology

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