bioRxiv2025
Macrophage activation is tightly coupled to cellular metabolism: classically activated pro-inflammatory (M1) macrophages rely on glycolysis and a disrupted tricarboxylic acid cycle, whereas alternatively activated (M2) macrophages depend on oxidative phosphorylation (OXPHOS) and fatty acid oxidation. Although mitochondria are central to this metabolic plasticity, it remains unclear whether mitochondrial dysfunction itself can dictate macrophage polarization. Using macrophage-specific OPA1 knockout mice, we investigated how mitochondrial dysfunction influences macrophage metabolism and immune homeostasis. Loss of OPA1 caused severe impairment of OXPHOS, reduced mitochondrial membrane potential, and a compensatory glycolytic shift, driving M0 and M2 macrophages toward an M1-like bioenergetic state. Integrative metabolomic and transcriptomic analyses revealed strong priming of OPA1-deficient macrophages towards classical activation, including accumulation of M1-associated metabolites (lactate, succinate, itaconate) and upregulation of NF-{kappa}B-driven and other inflammatory gene programs, resulting in increased secretion of IL-6 and TNF even in the absence of stimulation. Functionally, this metabolic shift primed non-activated and M2 macrophages toward partial M1 polarization with enhanced bactericidal capacity, while simultaneously suppressing M2-associated processes such as proliferation, efferocytosis, and expression of Arg1, CD206, and RELM. In vivo, OPA1{Delta}M mice displayed reduced peritoneal macrophage abundance, impaired self-renewal after IL-4 complex stimulation, and compensatory monocyte recruitment. The remaining macrophages exhibited increased MHCII and reduced RELM expression, consistent with partial M1 skewing and loss of alternative activation. These local alterations were mirrored systemically: blood profiling revealed enhanced T-cell activation and sex-specific remodeling of immune composition during inflammation and aging. Collectively, these findings demonstrate that mitochondrial dysfunction serves as a cell-intrinsic cue that primes macrophages toward a glycolytic, pro-inflammatory phenotype while constraining their M2 properties and proliferative capacities. This dual metabolic and functional rewiring highlights mitochondrial integrity as a pivotal determinant of macrophage immunometabolic identity and reveals how its disruption can reshape both local and systemic immune homeostasis.