Immunometabolic reprogramming in macrophages infected with active and dormant <i>Cryptococcus neoformans</i> : differential modulation of respiration, glycolysis, and fatty acid utilization

Dormancy is an adaptation in which cells reduce their metabolism, transcription, and translation to stay alive under stressful conditions, preserving the capacity to reactivate once the environment reverts to favorable conditions. Dormancy and reactivation of <i>Cryptococcus neoformans</i> (<i>Cn</i>) are closely linked to intracellular residency within macrophages. Our previous work showed that <i>in vitro</i> murine macrophages rely on the viable but not cultivable (VBNC-a dormancy phenotype) fungus from active <i>Cn</i>, with striking differences in immunometabolic gene expression. Here, we analyzed the influence of VBNC and active <i>Cn</i> on the immunometabolism of infected macrophages, combining metabolic gene expression, mitochondrial membrane potential (ΔΨm), oxygen consumption analysis, and uptake of glucose and fatty acids. The active fungus induced mitochondrial depolarization, and increased glycolysis and mitochondrial oxygen consumption. VBNC infection in bone marrow-derived macrophage (BMDM) caused an attenuated modification in mitochondrial metabolism. However, we found differences in BMDM infected with VBNC vs those infected with active fungus, where VBNC induced an increment in fatty acid uptake in M0 and M1 BMDM, measured by incorporation of BODIPY-palmitate, accompanied by an increase in expression of fatty acid transporters <i>Fabp1</i> and <i>Fabp4</i>. Overall, distinct fatty acid-related responses induced by VBNC and active <i>Cn</i> suggest different immunomodulatory reactions, depending on the microbial growth stage. We posit that, for VBNC, some of these macrophage metabolic responses reflect the establishment of prolonged microbial intracellular residency and possibly initial stages of granuloma formation.