Ready-to-Use-Type Lyophilized Lipid Nanoparticle Formulation for the Postencapsulation of Messenger RNA

Hiroki Tanaka; Shinya Hagiwara; Daiki Shirane; Takuma Yamakawa; Yuka Sato; Chika Matsumoto; Kota Ishizaki; Miho Hishinuma; Katsuyuki Chida; Kasumi Sasaki; Etsuo Yonemochi; Keisuke Ueda; Kenjirou Higashi; Kunikazu Moribe; Takashi Tadokoro; Katsumi Maenaka; Sakura Taneichi; Yuta Nakai; Kota Tange; Yu Sakurai; Hidetaka Akita

doi:10.1021/acsnano.2c10501

Ready-to-Use-Type Lyophilized Lipid Nanoparticle Formulation for the Postencapsulation of Messenger RNA

Hiroki Tanaka, Shinya Hagiwara, Daiki Shirane, Takuma Yamakawa, Yuka Sato, Chika Matsumoto, Kota Ishizaki, Miho Hishinuma, Katsuyuki Chida, Kasumi Sasaki, Etsuo Yonemochi, Keisuke Ueda, Kenjirou Higashi, Kunikazu Moribe, Takashi Tadokoro, Katsumi Maenaka, Sakura Taneichi, Yuta Nakai, Kota Tange, Yu SakuraiHidetaka Akita

PHA - Life and Pharmaceutical Science

Research output: Contribution to journal › Article › peer-review

Abstract

Based on the clinical success of an in vitro transcribed mRNA (IVT-mRNA) that is encapsulated in lipid nanoparticles (mRNA-LNPs), there is a growing demand by researchers to test whether their own biological findings might be applicable for use in mRNA-based therapeutics. However, the equipment and/or know-how required for manufacturing such nanoparticles is often inaccessible. To encourage more innovation in mRNA therapeutics, a simple method for preparing mRNA-LNPs is prerequisite. In this study, we report on a method for encapsulating IVT-mRNA into LNPs by rehydrating a Ready-to-Use empty freeze-dried LNP (LNPs(RtoU)) formulation with IVT-mRNA solution followed by heating. The resulting mRNA-LNPs(RtoU) had a similar intraparticle structure compared to the mRNA-LNPs prepared by conventional microfluidic mixing. In vivo genome editing, a promising application of these types of mRNA-LNPs, was accomplished using the LNPs(RtoU) containing co-encapsulated Cas9-mRNA and a small guide RNA.

Original language	English
Pages (from-to)	2588-2601
Number of pages	14
Journal	ACS Nano
Volume	17
Issue number	3
DOIs	https://doi.org/10.1021/acsnano.2c10501
Publication status	Published - 2023 Feb 14

Keywords

freeze-drying
gene delivery
gene editing
lipid nanoparticles
mRNA delivery

ASJC Scopus subject areas

Materials Science(all)
Engineering(all)
Physics and Astronomy(all)

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10.1021/acsnano.2c10501

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Tanaka, H., Hagiwara, S., Shirane, D., Yamakawa, T., Sato, Y., Matsumoto, C., Ishizaki, K., Hishinuma, M., Chida, K., Sasaki, K., Yonemochi, E., Ueda, K., Higashi, K., Moribe, K., Tadokoro, T., Maenaka, K., Taneichi, S., Nakai, Y., Tange, K., ... Akita, H. (2023). Ready-to-Use-Type Lyophilized Lipid Nanoparticle Formulation for the Postencapsulation of Messenger RNA. ACS Nano, 17(3), 2588-2601. https://doi.org/10.1021/acsnano.2c10501

Tanaka, H, Hagiwara, S, Shirane, D, Yamakawa, T, Sato, Y, Matsumoto, C, Ishizaki, K, Hishinuma, M, Chida, K, Sasaki, K, Yonemochi, E, Ueda, K, Higashi, K, Moribe, K, Tadokoro, T, Maenaka, K, Taneichi, S, Nakai, Y, Tange, K, Sakurai, Y & Akita, H 2023, 'Ready-to-Use-Type Lyophilized Lipid Nanoparticle Formulation for the Postencapsulation of Messenger RNA', ACS Nano, vol. 17, no. 3, pp. 2588-2601. https://doi.org/10.1021/acsnano.2c10501

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title = "Ready-to-Use-Type Lyophilized Lipid Nanoparticle Formulation for the Postencapsulation of Messenger RNA",

abstract = "Based on the clinical success of an in vitro transcribed mRNA (IVT-mRNA) that is encapsulated in lipid nanoparticles (mRNA-LNPs), there is a growing demand by researchers to test whether their own biological findings might be applicable for use in mRNA-based therapeutics. However, the equipment and/or know-how required for manufacturing such nanoparticles is often inaccessible. To encourage more innovation in mRNA therapeutics, a simple method for preparing mRNA-LNPs is prerequisite. In this study, we report on a method for encapsulating IVT-mRNA into LNPs by rehydrating a Ready-to-Use empty freeze-dried LNP (LNPs(RtoU)) formulation with IVT-mRNA solution followed by heating. The resulting mRNA-LNPs(RtoU) had a similar intraparticle structure compared to the mRNA-LNPs prepared by conventional microfluidic mixing. In vivo genome editing, a promising application of these types of mRNA-LNPs, was accomplished using the LNPs(RtoU) containing co-encapsulated Cas9-mRNA and a small guide RNA.",

keywords = "freeze-drying, gene delivery, gene editing, lipid nanoparticles, mRNA delivery",

author = "Hiroki Tanaka and Shinya Hagiwara and Daiki Shirane and Takuma Yamakawa and Yuka Sato and Chika Matsumoto and Kota Ishizaki and Miho Hishinuma and Katsuyuki Chida and Kasumi Sasaki and Etsuo Yonemochi and Keisuke Ueda and Kenjirou Higashi and Kunikazu Moribe and Takashi Tadokoro and Katsumi Maenaka and Sakura Taneichi and Yuta Nakai and Kota Tange and Yu Sakurai and Hidetaka Akita",

note = "Funding Information: H. Tanaka was supported by the JSPS KAKENHI [grant numbers 18K18377, 21K18035] and Kato Memorial Bioscience Foundation. H. Akita was supported by a JST CREST grant [grant number JPMJCR17H1], JSPS KAKENHI [grant numbers 20H00657, 21K18320], and the Asahi Glass Foundation. H.T. and H.A. were funded by NOF CORPORATION as a joint research. K. Maenaka was partly supported by Hokkaido University, Global Facility Center (GFC), Pharma Science Open Unit (PSOU), AMED [grant numbers JP21am0101093, JP22ama121037], JSPS KAKENHI [grant number JP20H05873], and Takeda Science Foundation. Publisher Copyright: {\textcopyright} 2023 American Chemical Society.",

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doi = "10.1021/acsnano.2c10501",

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T1 - Ready-to-Use-Type Lyophilized Lipid Nanoparticle Formulation for the Postencapsulation of Messenger RNA

AU - Tanaka, Hiroki

AU - Hagiwara, Shinya

AU - Shirane, Daiki

AU - Yamakawa, Takuma

AU - Sato, Yuka

AU - Matsumoto, Chika

AU - Ishizaki, Kota

AU - Hishinuma, Miho

AU - Chida, Katsuyuki

AU - Sasaki, Kasumi

AU - Yonemochi, Etsuo

AU - Ueda, Keisuke

AU - Higashi, Kenjirou

AU - Moribe, Kunikazu

AU - Tadokoro, Takashi

AU - Maenaka, Katsumi

AU - Taneichi, Sakura

AU - Nakai, Yuta

AU - Tange, Kota

AU - Sakurai, Yu

AU - Akita, Hidetaka

N1 - Funding Information: H. Tanaka was supported by the JSPS KAKENHI [grant numbers 18K18377, 21K18035] and Kato Memorial Bioscience Foundation. H. Akita was supported by a JST CREST grant [grant number JPMJCR17H1], JSPS KAKENHI [grant numbers 20H00657, 21K18320], and the Asahi Glass Foundation. H.T. and H.A. were funded by NOF CORPORATION as a joint research. K. Maenaka was partly supported by Hokkaido University, Global Facility Center (GFC), Pharma Science Open Unit (PSOU), AMED [grant numbers JP21am0101093, JP22ama121037], JSPS KAKENHI [grant number JP20H05873], and Takeda Science Foundation. Publisher Copyright: © 2023 American Chemical Society.

PY - 2023/2/14

Y1 - 2023/2/14

N2 - Based on the clinical success of an in vitro transcribed mRNA (IVT-mRNA) that is encapsulated in lipid nanoparticles (mRNA-LNPs), there is a growing demand by researchers to test whether their own biological findings might be applicable for use in mRNA-based therapeutics. However, the equipment and/or know-how required for manufacturing such nanoparticles is often inaccessible. To encourage more innovation in mRNA therapeutics, a simple method for preparing mRNA-LNPs is prerequisite. In this study, we report on a method for encapsulating IVT-mRNA into LNPs by rehydrating a Ready-to-Use empty freeze-dried LNP (LNPs(RtoU)) formulation with IVT-mRNA solution followed by heating. The resulting mRNA-LNPs(RtoU) had a similar intraparticle structure compared to the mRNA-LNPs prepared by conventional microfluidic mixing. In vivo genome editing, a promising application of these types of mRNA-LNPs, was accomplished using the LNPs(RtoU) containing co-encapsulated Cas9-mRNA and a small guide RNA.

AB - Based on the clinical success of an in vitro transcribed mRNA (IVT-mRNA) that is encapsulated in lipid nanoparticles (mRNA-LNPs), there is a growing demand by researchers to test whether their own biological findings might be applicable for use in mRNA-based therapeutics. However, the equipment and/or know-how required for manufacturing such nanoparticles is often inaccessible. To encourage more innovation in mRNA therapeutics, a simple method for preparing mRNA-LNPs is prerequisite. In this study, we report on a method for encapsulating IVT-mRNA into LNPs by rehydrating a Ready-to-Use empty freeze-dried LNP (LNPs(RtoU)) formulation with IVT-mRNA solution followed by heating. The resulting mRNA-LNPs(RtoU) had a similar intraparticle structure compared to the mRNA-LNPs prepared by conventional microfluidic mixing. In vivo genome editing, a promising application of these types of mRNA-LNPs, was accomplished using the LNPs(RtoU) containing co-encapsulated Cas9-mRNA and a small guide RNA.

KW - freeze-drying

KW - gene delivery

KW - gene editing

KW - lipid nanoparticles

KW - mRNA delivery

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ER -

Ready-to-Use-Type Lyophilized Lipid Nanoparticle Formulation for the Postencapsulation of Messenger RNA

Abstract

Keywords

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