Abstract
Purpose :
We previously reported that HSV-1 gK is involved in exacerbation of corneal disease following ocular challenge of gK-vaccinated mice. We have also shown that gK binds to signal peptide peptidase (SPP) and HSV-1 UL20, while UL20 binds to GODZ (zinc finger DHHC-type containing 3 (ZDHHC3) gene; also known as Golgi-specific DHHC zinc finger protein). The goal of this study was to map the gK binding region to SPP and UL-20 as well as mapping of UL-20 to GODZ in order to determine if blocking gK binding to SPP and UL20 using gK peptides and UL-20 peptides binding to GODZ (alone or in combination) will reduce HSV-1 infectivity in vitro and in vivo.
Methods :
The gK domain binding to SPP and UL20 as well as UL20 binding to GODZ were mapped using co-immunoprecipitation assays. Effect of these peptides on virus infectivity was determined by plaque assay, FACS and RT-PCR. The effect of ocular administration of selected peptides on ocular HSV-1 replication was evaluated in C57BL/6 mice by using standard plaque assays.
Results :
Previously we have shown that gK binds to SPP, UL20 binds to GODZ, and gK binds to UL20. Using overlapping gK and UL20 peptides, we screened the peptides which blocks SPP binding to gK, UL20 binding to GODZ, and gK binding to UL20, and tested the viral inhibition activities of the identified peptides in vitro. Our results also suggest a cocktail of peptides that are involved in blocking gK-SPP, UL20-GODZ and gK-UL20 is more efficient in reducing virus replication in HeLa, Vero, and RS cells than individual peptides corresponding to blocking of gK-SPP, UL20-GODZ, or gK-UL20 binding. Study is in progress to test this cocktail in vivo.
Conclusions :
Our results overall suggest that a cocktail of gK-UL20 peptides significantly reduced HSV-1 replication in vitro and in vivo. Thus, this cocktail of peptides may have therapeutic potential for control of ocular HSV-1 infections in vivo. As there is currently no effective treatment for HSV-1 recurrences, blocking SPP-gK-GODZ-UL20 interactions may represent a clinically effective and expedient target in developing HSV-1 therapeutics.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.