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Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method.

Min Tang¹, Ayane Sagawa¹, Nodoka Inoue¹

¹Department of Molecular Pharmaceutics, Hoshi University, 2-4-41 Ebara, Shinagawa, Tokyo 142-8501, Japan.

|April 28, 2023

Summary

The modified ethanol injection (MEI) method enhances messenger RNA (mRNA) lipoplex formation for improved in vitro and in vivo protein expression, showing promise for mRNA-based therapies and vaccinations.

Keywords:

delivery lipoplex mRNA mRNA vaccine

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Area of Science:

Biotechnology
Molecular Biology
Drug Delivery Systems

Background:

Messenger RNA (mRNA)-based therapies represent a significant advancement in vaccination and protein replacement for genetic disorders.
Previous research established a modified ethanol injection (MEI) method for effective small interfering RNA (siRNA) delivery.
This study explores the adaptation of the MEI method for mRNA delivery systems.

Purpose of the Study:

To investigate the efficacy of the modified ethanol injection (MEI) method in preparing messenger RNA (mRNA) lipoplexes.
To evaluate the in vitro and in vivo protein expression efficiency of mRNA lipoplexes generated using the MEI method.
To identify optimal lipid formulations for enhanced mRNA transfection.

Main Methods:

Screened six cationic lipids and three neutral helper lipids to create 18 distinct mRNA lipoplex formulations.
Incorporated polyethylene glycol-cholesteryl ether (PEG-Chol) into the lipoplex structures.
Assessed protein expression levels in cellular assays and in vivo studies in mice.

Main Results:

mRNA lipoplexes containing specific cationic lipids (DC-1-16 or TC-1-12) with DOPE and PEG-Chol demonstrated high protein expression in vitro.
Lipoplexes formulated with DC-1-16, DOPE, and PEG-Chol showed significant protein expression in mouse lungs and spleen following systemic administration.
Immunization with these lipoplexes resulted in elevated antigen-specific IgG1 antibody levels.

Conclusions:

The modified ethanol injection (MEI) method is effective for producing high-performance mRNA lipoplexes.
Optimized mRNA lipoplex formulations facilitate efficient protein expression both in vitro and in vivo.
The MEI method holds potential for improving the efficiency of mRNA-based therapeutics and vaccines.