Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method
Messenger RNA (mRNA)-based therapies are a novel class of therapeutics used in vaccination and protein replacement therapies for monogenic diseases. Previously, we developed a modified ethanol injection (MEI) method for small interfering RNA (siRNA) transfection, in which cationic liposome/siRNA com...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-04-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/15/4/1141 |
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| author | Min Tang Ayane Sagawa Nodoka Inoue Satomi Torii Kana Tomita Yoshiyuki Hattori |
| author_facet | Min Tang Ayane Sagawa Nodoka Inoue Satomi Torii Kana Tomita Yoshiyuki Hattori |
| author_sort | Min Tang |
| collection | DOAJ |
| description | Messenger RNA (mRNA)-based therapies are a novel class of therapeutics used in vaccination and protein replacement therapies for monogenic diseases. Previously, we developed a modified ethanol injection (MEI) method for small interfering RNA (siRNA) transfection, in which cationic liposome/siRNA complexes (siRNA lipoplexes) were prepared by mixing a lipid-ethanol solution with a siRNA solution. In this study, we applied the MEI method to prepare mRNA lipoplexes and evaluated the in vitro and in vivo protein expression efficiencies. We selected six cationic lipids and three neutral helper lipids to generate 18 mRNA lipoplexes. These were composed of cationic lipids, neutral helper lipids, and polyethylene glycol-cholesteryl ether (PEG-Chol). Among them, mRNA lipoplexes containing <i>N</i>-hexadecyl-<i>N</i>,<i>N</i>-dimethylhexadecan-1-aminium bromide (DC-1-16) or 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl) propan-2-yl) amino)-<i>N</i>,<i>N</i>,<i>N</i>-trimethyl-11-oxoundecan-1-aminium bromide (TC-1-12) with 1,2-dioleoyl-<i>sn</i>-glycero-3-phosphoethanolamine (DOPE) and PEG-Chol exhibited high protein expression in cells. Furthermore, mRNA lipoplexes composed of DC-1-16, DOPE, and PEG-Chol exhibited high protein expression in the lungs and spleen of mice after systemic injection and induced high antigen-specific IgG1 levels upon immunization. These results suggest that the MEI method can potentially increase the efficiency of mRNA transfection, both in vitro and in vivo. |
| first_indexed | 2024-03-11T04:38:46Z |
| format | Article |
| id | doaj.art-e78e8b743dbf408ebc012c94e6f1915b |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-11T04:38:46Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-e78e8b743dbf408ebc012c94e6f1915b2023-11-17T20:53:18ZengMDPI AGPharmaceutics1999-49232023-04-01154114110.3390/pharmaceutics15041141Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection MethodMin Tang0Ayane Sagawa1Nodoka Inoue2Satomi Torii3Kana Tomita4Yoshiyuki Hattori5Department of Molecular Pharmaceutics, Hoshi University, 2-4-41 Ebara, Shinagawa, Tokyo 142-8501, JapanDepartment of Molecular Pharmaceutics, Hoshi University, 2-4-41 Ebara, Shinagawa, Tokyo 142-8501, JapanDepartment of Molecular Pharmaceutics, Hoshi University, 2-4-41 Ebara, Shinagawa, Tokyo 142-8501, JapanDepartment of Molecular Pharmaceutics, Hoshi University, 2-4-41 Ebara, Shinagawa, Tokyo 142-8501, JapanDepartment of Molecular Pharmaceutics, Hoshi University, 2-4-41 Ebara, Shinagawa, Tokyo 142-8501, JapanDepartment of Molecular Pharmaceutics, Hoshi University, 2-4-41 Ebara, Shinagawa, Tokyo 142-8501, JapanMessenger RNA (mRNA)-based therapies are a novel class of therapeutics used in vaccination and protein replacement therapies for monogenic diseases. Previously, we developed a modified ethanol injection (MEI) method for small interfering RNA (siRNA) transfection, in which cationic liposome/siRNA complexes (siRNA lipoplexes) were prepared by mixing a lipid-ethanol solution with a siRNA solution. In this study, we applied the MEI method to prepare mRNA lipoplexes and evaluated the in vitro and in vivo protein expression efficiencies. We selected six cationic lipids and three neutral helper lipids to generate 18 mRNA lipoplexes. These were composed of cationic lipids, neutral helper lipids, and polyethylene glycol-cholesteryl ether (PEG-Chol). Among them, mRNA lipoplexes containing <i>N</i>-hexadecyl-<i>N</i>,<i>N</i>-dimethylhexadecan-1-aminium bromide (DC-1-16) or 11-((1,3-bis(dodecanoyloxy)-2-((dodecanoyloxy)methyl) propan-2-yl) amino)-<i>N</i>,<i>N</i>,<i>N</i>-trimethyl-11-oxoundecan-1-aminium bromide (TC-1-12) with 1,2-dioleoyl-<i>sn</i>-glycero-3-phosphoethanolamine (DOPE) and PEG-Chol exhibited high protein expression in cells. Furthermore, mRNA lipoplexes composed of DC-1-16, DOPE, and PEG-Chol exhibited high protein expression in the lungs and spleen of mice after systemic injection and induced high antigen-specific IgG1 levels upon immunization. These results suggest that the MEI method can potentially increase the efficiency of mRNA transfection, both in vitro and in vivo.https://www.mdpi.com/1999-4923/15/4/1141mRNAlipoplexdeliverymRNA vaccine |
| spellingShingle | Min Tang Ayane Sagawa Nodoka Inoue Satomi Torii Kana Tomita Yoshiyuki Hattori Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method Pharmaceutics mRNA lipoplex delivery mRNA vaccine |
| title | Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method |
| title_full | Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method |
| title_fullStr | Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method |
| title_full_unstemmed | Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method |
| title_short | Efficient mRNA Delivery with mRNA Lipoplexes Prepared Using a Modified Ethanol Injection Method |
| title_sort | efficient mrna delivery with mrna lipoplexes prepared using a modified ethanol injection method |
| topic | mRNA lipoplex delivery mRNA vaccine |
| url | https://www.mdpi.com/1999-4923/15/4/1141 |
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