Innovative γ-Oryzanol and KC2 Based Lipid Nanoparticles: OryKL Platform Provides Safe and Efficient In Vivo mRNA...

Area of Science:

• Biotechnology
• Materials Science
• Nanomedicine

Background:

• Current lipid nanoparticle (LNP) delivery systems for mRNA therapeutics face limitations including poor biocompatibility, immunogenicity, and cold-chain requirements.
• Improving LNP physicochemical and biological profiles is crucial for advancing mRNA nanotherapeutics for diverse disease treatments.

Purpose of the Study:

• To develop improved lipid nanoparticle (LNP) formulations for mRNA delivery by screening lipid mixtures.
• To identify a novel, biocompatible, and storage-stable LNP platform for enhanced mRNA delivery.

Main Methods:

• Systematic screening of a 20-component lipid mixture library, substituting ionizable lipids and sterol components.
• Characterization of lead LNP candidate (OryKL) nanostructures, mRNA encapsulation, and in vitro transfection efficiency.
• In vivo evaluation of OryKL in Ai9 reporter mice for mRNA delivery, biodistribution, toxicity, and inflammation, alongside stability studies in lyophilized form.

Main Results:

• A lead LNP candidate, OryKL, combining γ-oryzanol and DLin-KC2-DMA, demonstrated spherical nanostructures (∼150 nm), high mRNA encapsulation, and superior in vitro transfection efficiency compared to cholesterol controls.
• Intravenous administration of OryKL effectively delivered Cre recombinase mRNA to multiple organs in mice with no detectable toxicity or inflammation.
• Lyophilized OryKL with sucrose retained transfection potency for over 60 days, indicating ambient-stable storage capability.

Conclusions:

• γ-oryzanol serves as a viable and promising sterol alternative in LNP formulations.
• The OryKL platform offers a biocompatible, highly effective, and storage-stable solution for next-generation mRNA therapeutics.
• This study establishes a foundation for developing more clinically applicable mRNA delivery systems overcoming current limitations.