Challenges in Making and Using Lipid Nanoparticles for Gene Therapy, Including Skin Delivery, Cell Entry, Protein Interactions, and Machine Learning

Lipid nanoparticles (LNPs) are the primary delivery platform for nucleic acid therapeutics, yet their therapeutic performance is often assessed through indirect measurements.

• Progress in LNP development is hindered by a disconnect between physicochemical measurements and actual biological outcomes.
• A new antifouling coating has significantly reduced fouling in microfluidic channels, though its scalability is still unverified.
• Lyophilization remains a challenge, requiring re-optimization for each new lipid formulation, impacting clinical application.
• Current assessments of microneedle delivery focus on physicochemical integrity, lacking direct comparisons of endosomal escape capacity.
• Machine learning models for LNP formulation are constrained by biased training data rather than the algorithms themselves.
• Independent evaluations of endosomal escape efficiency indicate a low upper limit, with the reasons for this limitation still unclear.

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