Host-like Temperature Unlocks Glucose Uptake and Metabolism in Pathogenic Leptospira

Pathogenic Leptospira transitions between environmental reservoirs and mammalian hosts, expose the bacterium to distinct temperature conditions. Although Leptospira is generally considered to rely its growth primarily on long-chain fatty acids and to have limited capacity to use glucose, whether host-like temperature alters glucose-associated metabolic capacity remains unclear. Here, we used transcriptome-integrated genome-scale metabolic modeling to examine temperature-dependent metabolic states in pathogenic Leptospira. Models contextualized with transcriptomic data from cultures at 37{degrees}C predicted increased flux through glucose transport, glucose phosphorylation, and downstream glucose-associated reactions than it did at 29-30{degrees}C. Similar temperature-dependent predictions were obtained for another pathogenic Leptospira species under in vitro conditions. Consistent with these findings, analysis of leptospires cultivated intraperitoneally in dialysis membrane chambers also predicted enhanced glucose uptake and metabolism under host-temperature conditions. In vitro validation experiments showed increased bacterial uptake or accumulation of a fluorescent glucose analogue, elevated expression of candidate glucose transporter genes, and glucose-dependent enhancement of bacterial growth at 37{degrees}C. Treatment with a glucose degradation pathway inhibitor, 2-deoxy-D-glucose further supported a contribution of glucose-associated processes to proliferation at 37{degrees}C. Together, these findings indicate that pathogenic Leptospira displays condition-dependent glucose uptake and glucose-associated metabolic activity that become apparent at host-like temperature, opposed to glucose-incompetent in an environmental-like temperature. This prediction-driven framework refines the conventional view of Leptospira carbon metabolism and provides a basis for future studies of glucose-associated metabolic capacity during mammalian infection. ImportanceEnvironmental reservoirs play a central role in the transmission of many bacterial pathogens, yet the metabolic mechanisms that enable adaptation to both environmental and host-associated niches remain largely unknown. Using a prediction-driven framework that combines contextualized genome-scale metabolic modeling with multi-omics analyses and experimental validation, we discovered that pathogenic Leptospira activates glucose uptake and metabolism only under host-like conditions. This finding resolves a long-standing misconception arising from studies performed under conventional laboratory conditions and fundamentally revises our understanding of Leptospira physiology. More importantly, our study establishes an integrative systems biology strategy for uncovering condition-dependent metabolic traits that would be difficult to identify experimentally alone, with broad applicability to diverse environmentally transmitted pathogens.