Biomimetic noncationic lipid nanoparticles for mRNA delivery

Although the tremendous progress has been made for mRNA delivery based on classical cat-ionic carriers, the excess cationic charge density of lipids was necessary to compress mRNA through electrostatic interaction, and with it comes inevitably adverse events including the highly inflammatory and cytotoxic effects. How to develop the disruptive technologies to overcome cationic nature of lipids remains a major challenge for safe and efficient mRNA delivery. Here, we prepared noncationic thiourea lipids nanoparticles (NC-TNP) to com-press mRNA by strong hydrogen bonds interaction between thiourea groups of NC-TNP and the phosphate groups of mRNA, abandoning the hidebound and traditional electro-static force to construct mRNA-cationic lipids formulation. NC-TNP was a delivery system for mRNA with simple, convenient, and repeatable preparation technology and showed negligible inflammatory and cytotoxicity side effects. Furthermore, we found that NC-TNP could escape the recycling pathway to inhibit the egress of internalized nanoparticles from the intracellular compartment to the extracellular milieu which was a common fact in mRNA- LNP (lipid nanoparticles) formulation. Therefore, NC-TNP- encapsulated mRNA showed higher gene transfection efficiency in vitro and in vivo than mRNA-LNP formula-tion. Unexpectedly, NC-TNP showed spleen targeting delivery ability with higher accumu-lation ratio (spleen/liver), compared with traditional LNP. Spleen- targeting NC-TNP with mRNA exhibited high mRNA- encoded antigen expression in spleen and elicited robust immune responses. Overall, our work establishes a proof of concept for the construction of a noncationic system for mRNA delivery with good inflammatory safety profiles, high gene transfection efficiency, and spleen- targeting delivery to induce permanent and robust humoral and cell- mediated immunity for disease treatments.