Targeting Host Metabolic and Epigenetic Rewiring Blocks Lytic Gammaherpesvirus Production

Gammaherpesviruses are oncogenic viruses that reprogram host cell metabolism to support viral production. Among these, murine herpesvirus 68 (MHV-68) serves as a model system for studying lytic gammaherpesvirus infection and associated host cell changes. To characterize host transcriptional alterations induced throughout lytic gammaherpesvirus infection and identify novel host pathways that may be therapeutically targeted, we performed temporal bulk RNA-sequencing of mock- and MHV-68-infected NIH 3T3 cells at various timepoints throughout the lytic cycle. Our analysis revealed widespread and progressive host gene expression changes, including robust innate immune pathways and extensive remodeling of metabolic gene expression. We further identified a strong activation of the pentose phosphate pathway (PPP) genes, accompanied by increased abundance in PPP metabolic intermediates. Pharmacological inhibition of the PPP with 6-aminonicotinamide (6-AN) reduced infectious virus production. Moreover, at the intersection of metabolic and transcriptional reprogramming, we identified infection-associated gene expression changes in chromatin-modulating enzymes, including Tet2, and their metabolite co-factors, such as α-KG. Pharmacological inhibition of Ten-Eleven Translocation (TET) enzymatic activity led to a marked decrease in infectious MHV-68 production. Collectively, these findings define a novel metabolic-epigenetic crosstalk that supports productive gammaherpesvirus replication and identifies host pathways that can be targeted to treat lytic gammaherpesvirus infections.