Incorporation of endosomal escape peptides into mRNA-lipid nanoparticles to enhance intracellular protein expression
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Date
2024-07-05
Authors
Garrido Areal, Sergio
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Abstract
Lipid nanoparticles (LNPs) have emerged as the most successful and advanced vector for messenger RNA (mRNA) delivery, with applications in vaccines, cancer immunotherapy, enzyme replacement therapies, cellular reprogramming or genome editing. However, successful mRNA delivery still remains difficult, with the endosomal escape representing one of the most important challenges. The incorporation of cell-penetrating peptides (CPPs) into lipid nanoparticles represents a promising strategy to enhance endosomal escape and thus intracellular protein expression. In this study, we investigated the incorporation of LAH5, a histidine-rich amphiphilic peptide, into LNPs formulated with cholesterol or β-sitosterol. According to our findings, LAH5 addition on LNPs resulted in an increased size, reduced ζ-potential, and decreased mRNA encapsulation efficiency. The combination of LAH5 and cholesterol resulted in a higher protein expression, as well as an enhanced cellular uptake and endosomal escape compared to the base cholesterol LNP formulation. Conversely, the combination of LAH5 and β-sitosterol in the same ratios negatively impacted the high expression levels achieved by β-sitosterol-LNPs. Further research is necessary to determine how LAH5 is exactly incorporated into the LNPs and the underlying reasons for this differential behavior.
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Keywords
Lipid nanoparticles, mRNA, LAH5, Endosomal escape, Gal9 assay