Species-specific gene expression manipulation in humanized livers of chimeric mice via siRNA-encapsulated lipid nanoparticle treatment

Liver-humanized chimeric mice (PXB-mice) are widely utilized for predicting human pharmacokinetics (PK) and as human disease models. However, residual metabolic activity of mouse hepatocytes in chimeric mice can interfere with accurate human PK estimation. Lipid nanoparticle (LNP)-formulated small i...

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Main Authors: Kazuto Yamazaki, Kenji Kubara, Go Sugahara, Hiroki Muto, Masae Yamamoto, Yuji Mano, Kaoru Mitsuhashi, Chihiro Yamasaki, Yuji Ishida, Chise Tateno, Yuta Suzuki
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Molecular Therapy: Methods & Clinical Development
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Online Access:http://www.sciencedirect.com/science/article/pii/S2329050125000610
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author Kazuto Yamazaki
Kenji Kubara
Go Sugahara
Hiroki Muto
Masae Yamamoto
Yuji Mano
Kaoru Mitsuhashi
Chihiro Yamasaki
Yuji Ishida
Chise Tateno
Yuta Suzuki
author_facet Kazuto Yamazaki
Kenji Kubara
Go Sugahara
Hiroki Muto
Masae Yamamoto
Yuji Mano
Kaoru Mitsuhashi
Chihiro Yamasaki
Yuji Ishida
Chise Tateno
Yuta Suzuki
author_sort Kazuto Yamazaki
collection DOAJ
description Liver-humanized chimeric mice (PXB-mice) are widely utilized for predicting human pharmacokinetics (PK) and as human disease models. However, residual metabolic activity of mouse hepatocytes in chimeric mice can interfere with accurate human PK estimation. Lipid nanoparticle (LNP)-formulated small interfering RNA (siRNA) treatment makes it possible to eliminate the shortcomings of chimeras and create new models. Therefore, we aimed to create a new model in which siRNA for mouse cytochrome P450 oxidoreductase (Por) gene was encapsulated in LNP and administered to PXB-mice. We validated the siRNA-LNP system in PXB-mice, showing that a single intravenous injection of LNP-formulated mouse-specific siRNA against transthyretin (Ttr) knocked down Ttr expression in the liver and decreased plasma mouse TTR levels without affecting hepatic TTR expression and plasma human TTR levels. We produced mouse Por-specific siRNA with high in vitro silencing activity (siPOR(Mm)) and confirmed the efficient knockdown of Por expression in the livers of PXB-mice administered intravenously with LNP-encapsulated siPOR (siPOR(Mm)/LNP). siPOR(Mm)/LNP treatment suppressed 4′-hydroxywarfarin, making the S-warfarin PK profile in PXB-mice more similar to that in humans. Thus, mouse-specific siRNA-LNP is a simple system to control gene expression in the remaining mouse hepatocytes of PXB-mice and create more humanized and invaluable models based on PXB-mice.
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spelling doaj-art-65306dca2504448884ceea0aff34a2332025-08-20T02:29:24ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012025-06-0133210146610.1016/j.omtm.2025.101466Species-specific gene expression manipulation in humanized livers of chimeric mice via siRNA-encapsulated lipid nanoparticle treatmentKazuto Yamazaki0Kenji Kubara1Go Sugahara2Hiroki Muto3Masae Yamamoto4Yuji Mano5Kaoru Mitsuhashi6Chihiro Yamasaki7Yuji Ishida8Chise Tateno9Yuta Suzuki10Eisai Co., Ltd., Tsukuba Research Laboratories, 5-1-3, Tokodai, Tsukuba, Ibaraki 300-2635, Japan; Corresponding author: Kazuto Yamazaki, Ph.D., Eisai Co., Ltd., Tsukuba Research Laboratories, 5-1-3, Tokodai, Tsukuba, Ibaraki 300-2635, Japan.Eisai Co., Ltd., Tsukuba Research Laboratories, 5-1-3, Tokodai, Tsukuba, Ibaraki 300-2635, JapanResearch and Development Department, PhoenixBio Co., Ltd., 3-4-1, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, JapanEisai Co., Ltd., Tsukuba Research Laboratories, 5-1-3, Tokodai, Tsukuba, Ibaraki 300-2635, JapanEisai Co., Ltd., Tsukuba Research Laboratories, 5-1-3, Tokodai, Tsukuba, Ibaraki 300-2635, JapanEisai Co., Ltd., Tsukuba Research Laboratories, 5-1-3, Tokodai, Tsukuba, Ibaraki 300-2635, JapanEisai Co., Ltd., Tsukuba Research Laboratories, 5-1-3, Tokodai, Tsukuba, Ibaraki 300-2635, JapanResearch and Development Department, PhoenixBio Co., Ltd., 3-4-1, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, JapanResearch and Development Department, PhoenixBio Co., Ltd., 3-4-1, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, Japan; Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8553, JapanResearch and Development Department, PhoenixBio Co., Ltd., 3-4-1, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, Japan; Graduate School of Biomedical and Health Sciences, Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima 734-8553, Japan; School of Pharmaceutical Sciences, Wakayama Medical University, 25-1, Shichibancho, Wakayama 640-8156, Japan; Corresponding author: Chise Tateno, Ph.D., Research and Development Department, PhoenixBio Co., Ltd., 3-4-1, Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, Japan.Eisai Co., Ltd., Tsukuba Research Laboratories, 5-1-3, Tokodai, Tsukuba, Ibaraki 300-2635, JapanLiver-humanized chimeric mice (PXB-mice) are widely utilized for predicting human pharmacokinetics (PK) and as human disease models. However, residual metabolic activity of mouse hepatocytes in chimeric mice can interfere with accurate human PK estimation. Lipid nanoparticle (LNP)-formulated small interfering RNA (siRNA) treatment makes it possible to eliminate the shortcomings of chimeras and create new models. Therefore, we aimed to create a new model in which siRNA for mouse cytochrome P450 oxidoreductase (Por) gene was encapsulated in LNP and administered to PXB-mice. We validated the siRNA-LNP system in PXB-mice, showing that a single intravenous injection of LNP-formulated mouse-specific siRNA against transthyretin (Ttr) knocked down Ttr expression in the liver and decreased plasma mouse TTR levels without affecting hepatic TTR expression and plasma human TTR levels. We produced mouse Por-specific siRNA with high in vitro silencing activity (siPOR(Mm)) and confirmed the efficient knockdown of Por expression in the livers of PXB-mice administered intravenously with LNP-encapsulated siPOR (siPOR(Mm)/LNP). siPOR(Mm)/LNP treatment suppressed 4′-hydroxywarfarin, making the S-warfarin PK profile in PXB-mice more similar to that in humans. Thus, mouse-specific siRNA-LNP is a simple system to control gene expression in the remaining mouse hepatocytes of PXB-mice and create more humanized and invaluable models based on PXB-mice.http://www.sciencedirect.com/science/article/pii/S2329050125000610siRNAspecies specificitylipid nanoparticlechimeric micehumanized liverscytochrome P450 oxidoreductase
spellingShingle Kazuto Yamazaki
Kenji Kubara
Go Sugahara
Hiroki Muto
Masae Yamamoto
Yuji Mano
Kaoru Mitsuhashi
Chihiro Yamasaki
Yuji Ishida
Chise Tateno
Yuta Suzuki
Species-specific gene expression manipulation in humanized livers of chimeric mice via siRNA-encapsulated lipid nanoparticle treatment
Molecular Therapy: Methods & Clinical Development
siRNA
species specificity
lipid nanoparticle
chimeric mice
humanized livers
cytochrome P450 oxidoreductase
title Species-specific gene expression manipulation in humanized livers of chimeric mice via siRNA-encapsulated lipid nanoparticle treatment
title_full Species-specific gene expression manipulation in humanized livers of chimeric mice via siRNA-encapsulated lipid nanoparticle treatment
title_fullStr Species-specific gene expression manipulation in humanized livers of chimeric mice via siRNA-encapsulated lipid nanoparticle treatment
title_full_unstemmed Species-specific gene expression manipulation in humanized livers of chimeric mice via siRNA-encapsulated lipid nanoparticle treatment
title_short Species-specific gene expression manipulation in humanized livers of chimeric mice via siRNA-encapsulated lipid nanoparticle treatment
title_sort species specific gene expression manipulation in humanized livers of chimeric mice via sirna encapsulated lipid nanoparticle treatment
topic siRNA
species specificity
lipid nanoparticle
chimeric mice
humanized livers
cytochrome P450 oxidoreductase
url http://www.sciencedirect.com/science/article/pii/S2329050125000610
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