CD39 mRNA therapy attenuates localized acute inflammation: A novel anti-inflammatory strategy using cationic...

Messenger RNA (mRNA) therapeutics offer a promising strategy for treating inflammatory disease by enabling transient local expression of therapeutic proteins. CD39 (ectonucleoside triphosphate diphosphohydrolase-1) hydrolyzes pro-inflammatory ATP and ADP and plays a central role in localized immune regulation. We developed cationic nanoliposomes (NLps) for the delivery of CD39 mRNA and evaluated them in a murine model of localized inflammation induced by lipopolysaccharide and matrigel. Biodistribution studies showed substantial retention within the Matrigel matrix, with limited systemic distribution at 24 h. CD39 mRNA-NLps significantly reduced cellular infiltration at day 5, with decreased monocyte and macrophage staining via histological analysis. qPCR confirmed sustained local CD39 mRNA, while flow cytometry demonstrated increased CD39 protein staining in matrigel-derived immune cells, and phosphate release assays showed functional ectonucleotidase activity at the inflammatory site. Interleukin (IL)-6 levels were slightly reduced in matrigel extracts following treatment, supporting suppression of local inflammation. Hemocompatibility was confirmed using ex vivo in human blood and in vivo in treated mice, with liver enzymes and histological analysis supporting formulation's safety and favorable tolerability. These findings establish CD39 mRNA-NLps as a safe, effective strategy for a spatially confined mRNA-based approach to regulate local inflammation.