Since work has begun in February 2017, a shared MD/PhD student through the NIH-Oxford-Cambridge Scholars program has been recruited (Chad Coomer) and work has started on the topic. For much of the first 12 months, Chad worked on the following: 1) technology acquisition and pilot testing, and 2) biochemical reagent acquisition and pilot testing. He has now begun the experimental side of the project, crafting and testing hypotheses while making use of plasmid-encoded biosensors that measure intracellular metabolites. By studying biosensor activity following HIV exposure and during the specific stage of virus-cell fusion, he has found that glycolytic activity is positively associated with HIV entry into cells. Glycolytic activity is measured by a biosensor reporting the ratio of ATP to ADP. Additionally, the use of an inhibitor of glycolysis, 2-deoxyglucose, inhibits HIV-cell fusion. Interestingly, when the entry pathway used by HIV was modified through envelope glycoprotein pseudotyping, sensitivity to 2-deoxyglucose was altered. We are now exploring how engagement of CD4 and the HIV coreceptors, CXCR4 and CCR5, are linked to changes in intracellular metabolites. In the future, single-cell RNAseq will be introduced into the system in order to associate metabolite changes to changes in transcription, which will help identify the molecular basis for virus-induced metabolic disturbances. The focus on metabolites in this project makes it relevant to work being done in Project 1 (The Intersection between Cell-Intrinsic Innate Immunity and Metabolic Sensing), as well as to cancer biology.
