Synthetic polynucleotides as endosomolytic agents and bioenergy sources

Nucleotides (NTs), such as adenosine triphosphate (ATP) and guanosine triphosphate (GTP), are signaling and bioenergy molecules to mediate a range of cellular pathways. We recently reported their significant endosomolytic activity. To evaluate whether polymeric NTs keep endosomolytic and bioenergetic functions of NTs in drug delivery and cell survival, NTs were polymerized by a coupling reaction to form polynucleotides (pNTs: pATP and pGTP) with their molecular weights around 500 kDa. The cellular toxicity, indicated by IC50, of pNT was as low as that of corresponding monomeric NT. pNTs were degraded by an intracellular enzyme, alkaline phosphatase. Introduction of pNTs in a polycation-gene complex (polyplex) enhanced the extent of gene expression in cancerous, non-cancerous, and stem cells, up to 1500-fold higher than that of pNT-free polyplex. In addition, cells stored in a pATP solution resulted in a significantly higher survival rate (e.g., up to 20% increase) when exposed to low temperatures than pATP-free solution. The presence of pNT in polyplexes prevented the reduction of transfection efficiency induced by a low temperature. The findings in this study suggest that endosomolytic and bioenergetic pNTs serve as a non-toxic gene carrier component and protect cells from a cold shock or energy depletion.

Graphical abstractNon-toxic, enzymatically-degradable polynucleotides (pNTs) such as pATP and pGTP were synthesized. Bioenergy-producing pATP reduced a low temperature-induced cellular damage. pNT's endosomolytic activity in pDNA nanocomplexes improved gene expression in the cells cultured at 37 °C as well as 4 °C. Endosomolytic and bioenergetic functions of pNTs would be beneficial in drug delivery.Figure optionsDownload full-size imageDownload high-quality image (99 K)Download as PowerPoint slide