Histidine Decapeptide-Incorporating Lipid Nanoparticles with Low Ionizable Lipids Proportion for Efficient Small...

Area of Science:

• Biotechnology
• Nanomedicine
• Drug Delivery

Background:

• Lipid nanoparticles (LNPs) are key for delivering nucleic acids like small interfering RNA (siRNA).
• Diversifying LNP lipid composition is essential for improving clinical efficacy and overcoming current limitations in siRNA therapeutics.
• Efficient encapsulation of therapeutic payloads, including peptides, remains a challenge for conventional LNP formulations.

Purpose of the Study:

• To fabricate and characterize LNPs with varied lipid compositions for enhanced siRNA delivery.
• To investigate the impact of different mixing processes on LNP formulation and payload encapsulation.
• To develop a potent and stable LNP formulation with a reduced ionizable lipid proportion for improved siRNA therapeutics.

Main Methods:

• Fabrication of LNPs using pipette, vortex, and microfluidic mixing methods.
• Optimization of ionizable lipid proportion using a cost-efficient vortex-mixing approach.
• Incorporation of histidine decapeptide (His10) into optimized LNPs (LNP5) via microfluidic mixing to create LNP5H.

Main Results:

• Successful incorporation of siRNA and hydrophobic dyes, but not hydrophilic peptides, using pipette and vortex mixing.
• Optimized LNP5 formulation with a lower ionizable lipid proportion (27.72%) was developed.
• His10-incorporated LNP5 (LNP5H) demonstrated a 1.6-fold increase in gene silencing efficiency compared to conventional LNPs (cLNPs).
• LNP5H maintained siRNA potency for 4 weeks under cold storage conditions (-70 °C in 1% sucrose solution).

Conclusions:

• Potent LNP formulations (LNP5H) can be fabricated using a low proportion of ionizable lipids.
• Fast and straightforward fabrication processes, including vortex and microfluidic mixing, are suitable for LNP development.
• These novel LNPs offer a promising platform for efficient intracellular delivery of siRNA therapeutics.