Departmental Papers (Dental)
Herpes Simplex Virus-1 Infection in Human Primary Corneal Epithelial Cells is Blocked by a Stapled Peptide that Targets Processive DNA Synthesis
Date of this Version
Purpose: Acyclovir is most commonly used for treating ocular Herpes Keratitis, a leading cause of infectious blindness. However, emerging resistance to Acyclovir resulting from mutations in the thymidine kinase gene of Herpes Simplex Virus −1 (HSV-1), has prompted the need for new therapeutics directed against a different viral protein. One novel target is the HSV-1 Processivity Factor which is essential for tethering HSV-1 Polymerase to the viral genome to enable long-chain DNA synthesis. Methods: A series of peptides, based on the crystal structure of the C-terminus of HSV-1 Polymerase, were constructed with hydrocarbon staples to retain their alpha-helical conformation. The stapled peptides were tested for blocking both HSV-1 DNA synthesis and infection. The most effective peptide was further optimized by replacing its negative N-terminus with two hydrophobic valine residues. This di-valine stapled peptide was tested for inhibiting HSV-1 infection of human primary corneal epithelial cells. Results: The stapled peptides blocked HSV-1 DNA synthesis and HSV-1 infection. The unstapled control peptide had no inhibitory effects. Specificity of the stapled peptides was confirmed by their inabilities to block infection by an unrelated virus. Significantly, the optimized di-valine stapled peptide effectively blocked HSV-1 infection in human primary corneal epithelial cells with selectivity index of 11.6. Conclusions: Hydrocarbon stapled peptides that simulate the α-helix from the C-terminus of HSV-1 DNA polymerase can specifically block DNA synthesis and infection of HSV-1 in human primary corneal epithelial cells. These stapled peptides provide a foundation for developing a topical therapeutic for treating human ocular Herpes Keratitis. © 2020
DNA polymerase, Herpes keratitis, Herpes simplex virus-1, Hydrocarbon stapled peptide, Processivity factor, DNA, Epithelial Cells, Herpesvirus 1, Human, Humans, Keratitis, Herpetic, Peptides, aciclovir, DNA polymerase, genomic DNA, lactate dehydrogenase, peptide, recombinant protein, valine, virus DNA, DNA, peptide, absorption, alpha helix, amino acid substitution, amino terminal sequence, Article, carboxy terminal sequence, cell adhesion, cell viability, controlled study, cornea epithelium, crystal structure, DNA replication, DNA synthesis, drug potency, enzyme linked immunosorbent assay, epithelium cell, herpes simplex, herpes simplex keratitis, human, Human alphaherpesvirus 1, human cell, hydrogen bond, hydrophobicity, in vitro study, nonhuman, protein conformation, protein cross linking, protein protein interaction, reticulocyte lysate, selectivity index, solvation, Vaccinia virus, Vero cell line, virus replication, epithelium cell, genetics, herpes simplex keratitis
Guan, H., Nuth, N., Lee, V., Lin, C., Mitchell, C. H., Lu, W., Scott, R. W., Parker, M. H., Kulp, J. L., Reitz, A. B., & Ricciardi, R. P. (2021). Herpes Simplex Virus-1 Infection in Human Primary Corneal Epithelial Cells is Blocked by a Stapled Peptide that Targets Processive DNA Synthesis. Ocular Surface, 19 313-321. http://dx.doi.org/10.1016/j.jtos.2020.11.001
Date Posted: 10 February 2023
This document has been peer reviewed.