Metabolic Reprogramming and Tunneling Nanotubes Cooperate to Regulate HIV-1 Latency Reactivation

HIV-1 latency in myeloid cells remains a major obstacle to viral eradication. Here, we applied a systems biology approach combining transcriptomics, proteomics, metabolic modeling, targeted metabolite profiling, and advanced imaging to investigate metabolic alterations in a pre-monocytic latent cell model (U1). Upon latency reversal, we identified disrupted α-ketoglutarate (AKG) homeostasis driven by mitochondrial biogenesis and glutamine/glutamate metabolism, supporting energy production and M2-like macrophage polarization. Reporter metabolite analysis predicted cytoplasmic amino acid accumulation, and functional assays showed that tryptophan suppressed HIV reactivation by promoting mitochondrial and antioxidant metabolism. Additionally, we observed enhanced formation of tunneling nanotubes (TNTs), which facilitated intercellular transfer of mitochondria and viral components, potentially aiding viral persistence. Our study reveals cell-type–specific metabolic reprogramming and intercellular communication mechanisms underlying HIV-1 persistence. Our findings highlight the glutamine-AKG axis and TNTs as promising targets for strategies aimed at eliminating long-lived macrophage-associated HIV reservoirs.