Inhibition of PFKFB3 in Macrophages has a Dual Effect on Tumor-Regulating Lipid Metabolism

Colorectal cancer is the third most common cancer worldwide, making lymph node re-covery critical for treatment decisions and prognosis. Within the colorectal tumor mi-croenvironment, the metabolic programming of tumor-associated macrophages (TAMs) can drive both pro- and anti-tumor responses, yet the specific glycolytic pathways gov-erning their pro-metastatic conversion present promising therapeutic targets. This study investigated the role of glycolysis activating enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) in mediating TAM metabolic polarization, and its potential as a therapeutic target. PFKFB3 expression was found to be predominant in tumor-associated macrophages in CRC human tumor sam-ples. Lipidomic analysis performed by HPLC-MS/MS revealed that PFKFB3 inhibition altered glycerophospholipid metabolism (p=6.13E-10), and shifted TAMs toward sphingolipid-mediated immunosuppressive metabolism. PFKFB3 activity was associated with a specific reduction in asparagine availability, potentially pointing to a targeted reprogramming of amino acid metabolism supporting distinct macrophage functions under conditions of intra-tumoral metabolic stress. These findings highlight PFKFB3 as an essential metabolic regulator of TAMs pro-tumoral activity in CRC, particularly in colon cancer.