Cancer-associated fibroblast-derived exosomal NAT10 promotes the malignant progression of colon cancer through the USP36/HOXA7/HK2 axis

Background: CAFs drive CC progression via intercellular communication. Exosomes in CAF-mediated metabolic reprogramming and metastasis are underexplored. This study investigates how CAF-derived exosomes regulate CC's malignant phenotypes.Methods: Cell viability was assessed by a cell counting kit-8 assay, and proliferation via a 5-Ethynyl-2’-deoxyuridine incorporation assay. Transwell invasion and sphere formation assays analyzed invasive capacity and cancer stem-like cell properties. A tube formation assay determined angiogenic potential. Colorimetric assays measured metabolic alterations. Qrt-PCR, Western blotting, and immunohistochemistry detected mRNA and protein expression. AcRIP, RIP, Co-IP, ubiquitination, ChIP, and dual-luciferase reporter assays investigated molecule associations. A CC mouse model evaluated the impact of NAT10-deficient CAF-exosomes on tumor progression in vivo. HE staining assessed tumor pathological features, and non-invasive imaging monitored lung metastasis.Results: CAF-conditioned medium  enhanced CC cells' proliferation, invasion, sphere formation, angiogenesis, and glycolysis. It elevated NAT10 expression, which was upregulated in CC tissues and cell lines. Depletion of NAT10 in CAFs suppressed these malignant phenotypes. Mechanistically, NAT10 stabilized USP36 expression through acetylation. Knockdown of NAT10 inhibited malignant behaviors by modulating USP36 levels. USP36 stabilized HOXA7 protein via deubiquitination, and its depletion suppressed tumor progression by targeting HOXA7. HOXA7 transcriptionally activated  HK2 expression, establishing a NAT10-USP36-HOXA7-HK2 regulatory axis. CAF-derived exosomes transferred NAT10 to recipient cells, and exosomes from NAT10-deficient CAFs reduced tumor formation and lung metastasis in vivo.Conclusion: CAF-derived exosomal NAT10 promoted the malignant progression of CC by activating the USP36/HOXA7/HK2 axis. This finding not only elucidates a novel mechanism underlying CAF-exosome-mediated tumor promotion but also highlights potential clinical implications.