Extrahepatic Gene Editing In Vivo Using Organic Solvent-Free Lipid Nanoparticles

Area of Science:

• Biotechnology
• Nanomedicine
• Gene Therapy

Background:

• Targeted gene therapy offers potential for various diseases but faces challenges in RNA delivery, immune responses, and vector side effects.
• Current lipid nanoparticle formulations often rely on components like poly(ethylene glycol) (PEG) and ethanol, necessitating re-evaluation for improved safety and efficiency.

Purpose of the Study:

• To develop a novel, purely water-based lipid nanoparticle formulation for enhanced RNA delivery.
• To overcome limitations associated with conventional lipid nanoparticle formulations, including the elimination of critical components like PEG and ethanol.

Main Methods:

• A purely water-based formulation process for lipid nanoparticles (LNPs) was developed, initially using a stealth lipid containing poly(2-methyl-2-oxazoline) (PMeOx).
• The formulation was expanded to include approved lipids and evaluated for transfection efficiency in primary human immune cells.
• In vivo studies compared the biodistribution of these novel LNPs with conventional cholesterol-rich LNPs.

Main Results:

• The water-based LNP formulation demonstrated material efficiency, time savings, and high reproducibility.
• These nanoparticles effectively transfected primary human immune cells and delivered multiple nucleotides for CRISPR-Cas9 applications.
• In vivo studies revealed preferential transfection in extrahepatic tissues, unlike conventional liver-targeting LNPs.

Conclusions:

• A purely water-based LNP formulation offers a promising alternative for targeted gene therapy, addressing key delivery challenges.
• The developed nanoparticles show efficient transfection in immune cells and versatile nucleotide delivery for gene editing applications.
• The preferential extrahepatic targeting of these novel LNPs expands their potential therapeutic applications beyond the liver.