Autolysosomal degradation-inhibitory nanoparticles for enhanced mRNA escape and cytokine therapy

Lipid nanoparticles (LNPs), pivotal for mRNA therapeutics, face an underappreciated barrier of autolysosomal degradation besides endosomal sequestration. Herein, lipid-conjugated chloroquine lipidoids were synthesized and introduced into nanoparticles by replacing ionizable lipids to prepare autolysosomal degradation-inhibitory (AUDI) LNPs. With a conjugated lipid consisting of 12 carbon atoms and a 30% substitution rate, AUDI-LNPs achieved optimal performance. Furthermore, the AUDI technology improved the delivery efficiency in vitro and in vivo of LNPs by approximately 5-fold and was compatible with various LNP formulations, cell lines, and mRNA types. Overall, the incorporation of autophagy inhibitor into LNPs is intended to prevent premature degradation, and AUDI-LNPs present a universal and plug-and-play strategy to circumvent autolysosomal degradation barrier for enhancing mRNA escape and treatment, providing new insights into future gene therapy and vaccine development with maintained safety margin.