Glutamine-Functionalized mRNA Lipid Nanoparticles Block HIF-1α-Dependent Glycolytic Adaptation to Sensitize Azelaic Acid Therapy in Colorectal Cancer

Metabolic plasticity limits mitochondria-directed cancer therapy by enabling glycolytic compensation during respiratory stress. Our previous nano-energy interference (NEI) platform showed that coupling azelaic acid (AZA)-induced mitochondrial disruption with restriction of glucose utilization suppresses colorectal cancer (CRC), but the upstream program coordinating this adaptive response remained unresolved. Here, we identify HIF-1α-dependent glycolytic adaptation as a key response to AZA-induced respiratory inhibition and develop glutamine-functionalized lipid nanoparticles (G-LNPs) to deliver dominant-negative HIF-1α (dnHIF-1α) mRNA. G-LNPs were approximately 105 nm in diameter, achieved >90% mRNA encapsulation, and enhanced uptake and functional reporter mRNA expression in ASCT2-expressing MC38 cells in a GPNA-sensitive manner. AZA suppressed mitochondrial respiration while activating HIF-responsive transcription, glycolytic activity, lactate production, and GLUT1, HK2, LDHA, and PDK1 expressions. G-LNP-delivered dnHIF-1α blocked this compensatory response without materially intensifying respiratory inhibition, thereby deepening ATP depletion and producing a distinct treatment-associated metabolomic state. Following treatment withdrawal and equal viable cell replating, the combination caused persistent impairment of regrowth and clonogenic capacity. In subcutaneous MC38 tumors, combined treatment produced the greatest tumor growth inhibition without overt systemic toxicity. Together, these findings support transient disruption of HIF-dependent metabolic adaptation as a strategy to convert partially recoverable mitochondrial stress into a durable loss of CRC cell fitness.