A naturally derived lipopeptide lipid nanoparticle platform enabling multiple nucleic acids delivery

Area of Science:

• Biotechnology
• Nanomedicine
• Gene Therapy

Background:

• Lipid nanoparticles (LNPs) are crucial for nucleic acid delivery but face challenges in tissue specificity and toxicity.
• Achieving targeted delivery and tunable properties remains essential for effective LNP-based therapeutics.

Purpose of the Study:

• To develop a novel, high-performance LNP platform using arginine-histidine peptide-dioleoylphosphatidylethanolamine (RmHnC-DOPE) lipopeptides.
• To engineer and evaluate LNP variants for optimized payload-specific and tissue-selective nucleic acid delivery.
• To assess the efficacy, targeting capabilities, and safety of the novel LNP platform compared to benchmarks.

Main Methods:

• Systematic optimization of arginine-to-histidine ratios in RmHnC-DOPE lipopeptides to create R3H7C-DOPE, R4H6C-DOPE, and R5H5C-DOPE variants.
• In vitro evaluation of gene silencing, transfection efficiency, and cell viability for different LNP formulations.
• In vivo studies assessing hepatic and extrahepatic targeting, gene expression, and therapeutic efficacy (e.g., PCSK9 suppression).
• Comprehensive safety and biocompatibility assessments, including survival rates and organ function monitoring.

Main Results:

• R4H6C-DOPE and R5H5C-DOPE showed significantly higher siRNA gene silencing efficiency (85.1% and 89.5%) than SM-102 LNPs (67.4%), with >99% cell viability.
• R5H5C-DOPE achieved high mRNA (74.8%) and plasmid DNA (pDNA) (92.1%) transfection efficiency, while R3H7C-DOPE demonstrated an 11-fold increase in pDNA expression.
• In vivo studies confirmed R3H7C-DOPE's hepatic targeting and superior PCSK9 suppression, whereas R5H5C-DOPE achieved >90% pulmonary localization, indicating effective extrahepatic targeting.
• RmHnC-DOPE formulations exhibited exceptional biocompatibility with 100% survival rates, contrasting with severe toxicity observed with SM-102 LNPs.

Conclusions:

• The novel RmHnC-DOPE LNP platform offers tunable properties for precise tissue targeting and efficient nucleic acid delivery.
• Engineered LNP variants provide superior efficacy and safety profiles for diverse gene therapy applications, including hepatic and extrahepatic targets.
• This platform presents a clinically viable solution for advancing nucleic acid-based therapeutics with enhanced biocompatibility and targeting precision.