title: Identification of a new class of small molecules that efficiently reactivate latent Epstein-Barr Virus.
abstract: Epstein-Barr Virus (EBV) persists as a latent infection in many lymphoid and epithelial malignancies, including Burkitt's lymphomas, nasopharyngeal carcinomas, and gastric carcinomas. Current chemotherapeutic treatments of EBV-positive cancers include broad-spectrum cytotoxic drugs that ignore the EBV-positive status of tumors. An alternative strategy, referred to as oncolytic therapy, utilizes drugs that stimulate reactivation of latent EBV to enhance the selective killing of EBV-positive tumors, especially in combination with existing inhibitors of herpesvirus lytic replication, like Ganciclovir (GCV). At present, no small molecule, including histone deacetylase (HDAC) inhibitors, have proven safe or effective in clinical trials for treatment of EBV-positive cancers. Aiming to identify new chemical entities that induce EBV lytic cycle, we have developed a robust high-throughput cell-based assay to screen 66,840 small molecule compounds. Five structurally related tetrahydrocarboline derivatives were identified, two of which had EC50 measurements in the range of 150-170 nM. We show that these compounds reactivate EBV lytic markers ZTA and EA-D in all EBV-positive cell lines we have tested independent of the type of latency. The compounds reactivate a higher percentage of latently infected cells than HDAC inhibitors or phorbol esters in many cell types. The most active compounds showed low toxicity to EBV-negative cells but were highly effective at selective cell killing of EBV-positive cells when combined with GCV. We conclude that we have identified a class of small molecule compounds that are highly effective at reactivating latent EBV infection in a variety of cell types and show promise for lytic therapy in combination with GCV.
authors.affiliation: The Wistar Institute , Philadelphia, Pennsylvania 19104, United States.
authors.forename: Nadezhda
authors.initials: N
authors.lastname: Tikhmyanova
authors.forename: David C
authors.initials: DC
authors.lastname: Schultz
authors.forename: Theresa
authors.initials: T
authors.lastname: Lee
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Salvino
authors.forename: Paul M
authors.initials: PM
authors.lastname: Lieberman
mesh_terms: D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004854:Herpesvirus 4, Human / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D057166:High-Throughput Screening Assays; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics; D015394:Molecular Structure; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D015534:Trans-Activators / Q000235:genetics; D014775:Virus Activation / Q000187:drug effects*; D017735:Virus Latency / Q000187:drug effects*; D014779:Virus Replication / Q000187:drug effects*
chemical_list: C063676:BZLF1 protein, Herpesvirus 4, Human; D017874:Immediate-Early Proteins; D054852:Small Molecule Libraries; D015534:Trans-Activators
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