Determining the Role of vGPCR in Modulating Host Cell Transcription and Metabolism
By: Osvaldo Kevin Moreno
Department: Cellular & Molecular Biology
Faculty Advisors: Dr. Erica L. Sanchez, Dr. Scott Roy, and Dr. Nicole Salazar Velmeshev
Kaposi Sarcoma-associated Herpesvirus (KSHV) is a small etiologic agent that can induce angiogenesis in endothelial cells, and lead to Kaposi Sarcoma (KS). KSHV has two life cycles: the latent, a more dormant stage of infection, and the lytic stage, in which virus is actively produced. Viral G Protein-Coupled Receptor (vGPCR) is a lytic signaling protein that has been connected to modulating host cell transcription and paracrine signaling. Our previous work identified the host cell metabolism to be crucial for KSHV infection, but the mechanism by which vGPCR modulates cellular metabolism has yet to be discovered. We hypothesize that vGPCR signaling dramatically alters the host transcriptome and metabolism. To test our hypothesis, we have successfully overexpressed vGPCR protein in mammalian cells, which was confirmed through western blots and immunofluorescence assays. Here we demonstrate that vGPCR expression can alter host cell transcription, as seen by the upregulation of the host gene cyclooxygenase-2 (COX2). Additionally, we are performing RNA sequencing analysis to obtain a host cell transcriptomic profile for vGPCR-expressing cells compared to empty vector expressing cells. This approach will identify changes in the transcript levels of key metabolic targets downstream of vGPCR signaling. We expect the transcriptome profile to show an increase expression in targets like Hypoxia-inducible factor-1 and Hexokinase-2, two targets critical for glucose metabolism that have been previously identified to be increased during KSHV infection. These experiments will elucidate the mechanism by which vGPCR modulates metabolic gene expression and cellular metabolism, helping lay the foundation to develop antiviral therapeutics.