A novel lipopolyplex platform for dual mRNA delivery via core- and surface-loading

Area of Science:

• Biotechnology
• Nanomedicine
• Polymer Science

Background:

• Recent approvals of nanoparticulate nucleotide therapies highlight delivery system advancements.
• Gene therapies like CRISPR/Cas9 and cancer immunotherapy require co-delivery of multiple nucleotides.
• Current lipid nanoparticles face challenges in co-encapsulation and managing nucleotide interactions.

Purpose of the Study:

• To design a core-shell system for separate encapsulation of two nucleotides.
• To develop a two-step formulation process for advanced nucleotide delivery.
• To create lipopolyplexes (LPPs) overcoming lipid nanoparticle limitations.

Main Methods:

• Synthesized 18 distinct core compositions by combining six cationic and three anionic polymers.
• Screened formulations to identify potent lipopolyplexes (LPPs).
• Evaluated LPPs for transfection efficiency, expression kinetics, storage stability, and nebulization performance.

Main Results:

• Identified poly-L-arginine and poly-L-glutamic acid as the optimal combination for LPPs.
• Achieved precise co-delivery of two model messenger RNAs (mRNAs) in a controlled ratio.
• Demonstrated that surface-loaded mRNA exhibited superior transfection efficiency and shear resistance compared to core-loaded mRNA.

Conclusions:

• Developed a novel core-shell LPP system for controlled co-delivery of multiple nucleotides.
• The poly-L-arginine/poly-L-glutamic acid LPP shows significant potential for advanced therapeutic applications.
• mRNA localization within LPPs critically impacts transfection efficiency and stability, especially under nebulization.