Use of a multi-omic approach to drug discovery for the treatment of HSV-1 infection

Abstract

Purpose : New treatments are needed to reduce blindness due to herpes simplex virus type-1 (HSV-1) infection of the eye. While all ocular tissues can be involved, involvement of the cornea is more common. Since the virus is known to modulate transcriptomic and proteomic profile of the host, we used a multi-omic approach to validate new targets in the corneal epithelium and identify a new investigational drug that more effectively curbs ocular HSV-1.

Methods : In this study, we used transcriptomic and proteomic methods to analyze differentially regulated pathways during HSV-1 infection in primary human corneal cells. Various analyses were conducted to reveal pro-viral pathways and drugs that target them. Immunoblotting and immunofluorescence assays were performed to confirm our findings in vitro and in vivo murine models of ocular infection to determine antiviral activity of newly discovered antiviral drugs. Synergy studies were conducted with acyclovir to determine if our newly discovered drug can provide improved benefit. Development of antiviral resistance was determined by growing the virus in growth media containing low drug concentration for multiple generations.

Results : Our studies revealed that cyclin dependent kinase activity mediated by E2F transcription is a vital component of HSV-1 infection. CDKs are crucial for HSV-1 replication in cornea and their silencing leads to loss of infection. Utilizing CMAP analysis, MD-simulation and kinase activity studies, we discovered a previously unknown CDK inhibitor that directly binds and inhibits these pro-viral factors to show excellent antiviral efficacy. Finally, through extensive in vitro and in vivo studies, we show that our novel drug can not only improve acyclovir’s antiviral activity even against clinical acyclovir-resistant strains of HSV, but also does not allow the virus to develop antiviral resistance.

Conclusions : Our studies reveal the importance of antiviral discovery and development in tissue specific environments.Although previously studies have shown the importance of CDKs in modulating HSV-1 infection,their predominance in the corneal tissue has remained unknown until now. Our studies reveal that combination therapies targeting the CDKs can significantly inhibit HSV-1 replication and drugs which target these factors can improve the efficacy of acyclovir against resistant viral strains.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.