m6A-Dependent Upregulation of PFKFB3 Drives Macrophage-Mediated Inflammation in Necrotizing Enterocolitis

Area of Science:

• Epigenetics
• Metabolic pathways
• Inflammatory diseases

Background:

• Necrotizing enterocolitis (NEC) is a severe gastrointestinal disease in neonates.
• The role of epigenetic modifications, specifically N6-methyladenosine (m6A) methylation, in NEC pathogenesis is not fully understood.
• 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (PFKFB3) is a key regulator of glycolysis.

Purpose of the Study:

• To investigate the role of m6A methylation in regulating PFKFB3 expression in NEC.
• To explore the therapeutic potential of targeting PFKFB3 in NEC.

Main Methods:

• Analysis of m6A methylation in human NEC tissues.
• In vivo and in vitro NEC models (lipopolysaccharide-hypoxia-cold stress, THP-1-differentiated macrophages).
• Assessment of PFKFB3 expression, glycolytic flux, reactive oxygen species (ROS) production, and macrophage polarization.
• Pharmacological inhibition of PFKFB3.

Main Results:

• Reduced m6A methylation of PFKFB3 mRNA was observed in human NEC tissues, leading to increased PFKFB3 stability and protein levels.
• PFKFB3-driven glycolysis promoted M1 macrophage polarization via ROS accumulation, exacerbating intestinal inflammation in NEC models.
• Pharmacological inhibition of PFKFB3 significantly reduced ROS, macrophage infiltration, and mucosal injury in NEC models.

Conclusions:

• A metabolic-epigenetic axis involving reduced m6A methylation of PFKFB3 mRNA contributes to NEC pathogenesis.
• PFKFB3 inhibition represents a promising therapeutic strategy for mitigating inflammation and tissue damage in NEC.
• This study provides the first evidence of an m6A-mediated mechanism in NEC and highlights PFKFB3 as a novel therapeutic target.