Resolving the mRNA Encapsulation-Release Trade-off via Compensatory Forces in Engineered Ionizable Lipids

Area of Science:

• Biotechnology
• Materials Science
• Molecular Biology

Background:

• Messenger RNA (mRNA) delivery via lipid nanoparticles (LNPs) faces a key challenge: balancing stable encapsulation with efficient intracellular release.
• This trade-off limits the therapeutic potential of mRNA-based treatments.

Purpose of the Study:

• To engineer LNPs that optimize mRNA encapsulation and intracellular release through compensatory force engineering.
• To develop a computational-experimental framework for designing ionizable lipids (ILs) with specific short-range interaction motifs.

Main Methods:

• Developed a "contact number" metric using a computational-experimental framework to guide LNP design.
• Incorporated short-range interaction motifs (urea, carbamate) into ionizable lipid structures.
• Evaluated LNP performance in mRNA translation, T cell responses, tumor inhibition, and gene editing efficiency.

Main Results:

• Engineered LNPs (OT13-LNPs) demonstrated optimal mRNA encapsulation and enhanced endosomal escape, leading to improved mRNA translation.
• OT13-LNPs showed a 1.7-fold increase in antigen-specific T cell responses and 77.9% tumor inhibition in melanoma models.
• In hepatic gene editing, OT13-LNPs achieved comparable on-target editing efficiency to commercial LNPs but a significantly stronger silencing effect (>90% vs. ~58% TTR reduction).

Conclusions:

• Compensatory force engineering offers a promising strategy for developing next-generation mRNA therapeutics.
• The developed LNP system shows potential for applications in oncology, gene editing, and infectious diseases.