Escobar, ThelmaJones, Chloe2026-02-052026-02-052026-02-052025Jones_washington_0250E_29093.pdfhttps://hdl.handle.net/1773/55145Thesis (Ph.D.)--University of Washington, 2025The human gammaherpesviruses (γHV) Kaposi Sarcoma-Associated Virus (KSHV) and Epstein Barr Virus (EBV) are two of the seven known human oncogenic viruses. KSHV infects around 5% of the population and causes cancers such as Kaposi Sarcoma in immunodeficient individuals, whereas EBV infects >90% of the population and is linked to lymphomas and, more recently, multiple sclerosis. Despite their impact on human health, there are currently no drugs to treat either virus. This may in part be due to difficulties studying the lytic phase in the human γHVs; KSHV and EBV both immediately enter latency upon initial infection. A better understanding of what processes occur during the lytic phase, the part of the viral lifecycle during which the virus actively replicates, may offer new insights on potentially druggable pathways. MHV-68 is a murine γHV that immediately enters the lytic phase upon primary infection and shares ~80% sequence similarity with the human γHVs, thus making it a good model for studying the lytic phase in the human γHVs. Surprisingly, there have been no prior studies of the host transcriptome during viral infection of NIH3T3 cells, one of the most common models for the study of MHV-68. In this body of work, I analyze bulk RNA-seq data from MHV-68 infected NIH3T3 cells with the goal of identifying potential druggable pathway(s). In taking a two-pronged approach in which I assess the most differentially expressed metabolic and epigenomic genes/pathways, I present transcriptomic evidence for increased redox stress in MHV-68 infected NIH3T3 cells, as well as evidence for perturbations in chromatin biology. My analysis indicated an increase in transcripts encoding components of the mitochondrial electron transport chain, glutathione synthesis, and NADPH-producing pathways, collectively suggesting increased oxidative stress over the course of infection. Notable changes in chromatin biology include upregulation of the expression of DNA demethylases and the DNA damage response chaperone DAXX. Additionally, canonical histone H2A was upregulated, while histone variant H2AX was downregulated late in lytic infection. Ultimately, I found that inhibition of the DNA demethylating enzymes Ten-eleven translocase 1 and 2 (Tet1 and Tet2) diminishes MHV-68 replication in this system. The mechanism behind how Tet1/2 inhibition reduces viral titer remains to be explored.application/pdfen-USCC BYchromatinferroptosisgammaherpesvirusmetabolismMHV-68TETBiochemistryVirologyBiological chemistryThesis