A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes
In this study, we explore the therapeutic feasibility of globotriaosylceramide (Gb3) synthase (A4GALT)-specific siRNA-loaded polyhistidine (pHis)-incorporated lipid nanoparticles (HLNPs) for Fabry disease (FD). HLNPs were developed to deliver siRNAs targeting A4GALT using a microfluidic device, with...
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Elsevier
2025-06-01
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| Series: | Molecular Therapy: Nucleic Acids |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253125001271 |
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| author | Yoo Jin Shin Hanbi Lee Xianying Fang Sheng Cui Sun Woo Lim Kang In Lee Jae Young Lee Hong Lim Kim Yuna Oh Can Li Chul Woo Yang Gayeon You Hyeondo Lee Hyejung Mok Byung Ha Chung |
| author_facet | Yoo Jin Shin Hanbi Lee Xianying Fang Sheng Cui Sun Woo Lim Kang In Lee Jae Young Lee Hong Lim Kim Yuna Oh Can Li Chul Woo Yang Gayeon You Hyeondo Lee Hyejung Mok Byung Ha Chung |
| author_sort | Yoo Jin Shin |
| collection | DOAJ |
| description | In this study, we explore the therapeutic feasibility of globotriaosylceramide (Gb3) synthase (A4GALT)-specific siRNA-loaded polyhistidine (pHis)-incorporated lipid nanoparticles (HLNPs) for Fabry disease (FD). HLNPs were developed to deliver siRNAs targeting A4GALT using a microfluidic device, with pHis aiding in endosome escape. The therapy was tested on GLA-knockout human-induced pluripotent-stem-cell-derived endothelial cells (GLA-KO-hiPSC-ECs) and podocytes (GLA-KO-hiPSC-PCs). GLA-KO-hiPSCs-ECs or -PCs, upon differentiation, were treated with A4GALT-siRNA-HLNP. Successful intracellular uptake of A4GALT-siRNA-HLNP was confirmed through fluorescence and electron microscopy in both cell types. A4GALT-siRNA-HLNP treatment confirmed both cell types’ stability at 5 μg/mL. Increased Gb3 deposition and zebra body formation were detected in both cell types, but A4GALT-siRNA-HLNP treatment attenuated these FD phenotypes, demonstrating reduced expression of A4GALT through western blot analysis. RNA sequencing analysis revealed that the expression of transcripts associated with FD was restored by A4GALT-siRNA-HLNP treatment in GLA-KO-hiPSCs-ECs, whereas in GLA-KO-hiPSCs-PCs, this effect was relatively less pronounced. Suppression of A4GALT via siRNA/HLNP treatment significantly rescued FD phenotypes especially in EC, presenting a novel therapeutic approach for FD. |
| format | Article |
| id | doaj-art-2ca493d262b04dc696e1ef7fae2d8615 |
| institution | DOAJ |
| issn | 2162-2531 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj-art-2ca493d262b04dc696e1ef7fae2d86152025-08-20T03:07:24ZengElsevierMolecular Therapy: Nucleic Acids2162-25312025-06-0136210257310.1016/j.omtn.2025.102573A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytesYoo Jin Shin0Hanbi Lee1Xianying Fang2Sheng Cui3Sun Woo Lim4Kang In Lee5Jae Young Lee6Hong Lim Kim7Yuna Oh8Can Li9Chul Woo Yang10Gayeon You11Hyeondo Lee12Hyejung Mok13Byung Ha Chung14Transplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea; Division of Nephrology, Department of Internal Medicine, Seoul St. Mary’s Hospital, The College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaR&D Center, ToolGen, Inc., Seoul 07789, Republic of KoreaR&D Center, ToolGen, Inc., Seoul 07789, Republic of Korea; Department of Anatomy, Ajou University School of Medicine, Suwon 16499, KoreaIntegrative Research Support Center, Laboratory of Electron Microscope, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaIntegrative Research Support Center, Laboratory of Electron Microscope, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaDepartment of Nephrology, Yanbian University Hospital, Yanji, ChinaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea; Division of Nephrology, Department of Internal Medicine, Seoul St. Mary’s Hospital, The College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaDepartment of Bioscience and Biotechnology, Konkuk University, Seoul 05029, Republic of KoreaDepartment of Bioscience and Biotechnology, Konkuk University, Seoul 05029, Republic of KoreaDepartment of Bioscience and Biotechnology, Konkuk University, Seoul 05029, Republic of Korea; Corresponding author: Hyejung Mok, Ph.D., Department of Bioscience and Biotechnology, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea.Transplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea; Division of Nephrology, Department of Internal Medicine, Seoul St. Mary’s Hospital, The College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea; Corresponding author: Byung Ha Chung M.D., Ph.D., Transplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul, Korea; Division of Nephrology, Department of Internal Medicine, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, Seoul, South Korea.In this study, we explore the therapeutic feasibility of globotriaosylceramide (Gb3) synthase (A4GALT)-specific siRNA-loaded polyhistidine (pHis)-incorporated lipid nanoparticles (HLNPs) for Fabry disease (FD). HLNPs were developed to deliver siRNAs targeting A4GALT using a microfluidic device, with pHis aiding in endosome escape. The therapy was tested on GLA-knockout human-induced pluripotent-stem-cell-derived endothelial cells (GLA-KO-hiPSC-ECs) and podocytes (GLA-KO-hiPSC-PCs). GLA-KO-hiPSCs-ECs or -PCs, upon differentiation, were treated with A4GALT-siRNA-HLNP. Successful intracellular uptake of A4GALT-siRNA-HLNP was confirmed through fluorescence and electron microscopy in both cell types. A4GALT-siRNA-HLNP treatment confirmed both cell types’ stability at 5 μg/mL. Increased Gb3 deposition and zebra body formation were detected in both cell types, but A4GALT-siRNA-HLNP treatment attenuated these FD phenotypes, demonstrating reduced expression of A4GALT through western blot analysis. RNA sequencing analysis revealed that the expression of transcripts associated with FD was restored by A4GALT-siRNA-HLNP treatment in GLA-KO-hiPSCs-ECs, whereas in GLA-KO-hiPSCs-PCs, this effect was relatively less pronounced. Suppression of A4GALT via siRNA/HLNP treatment significantly rescued FD phenotypes especially in EC, presenting a novel therapeutic approach for FD.http://www.sciencedirect.com/science/article/pii/S2162253125001271MT: Oligonucleotides: Therapies and ApplicationsFabry diseaseA4GALT siRNApolyhistidine-incorporating lipid nanoparticlehuman-induced pluripotent-stem-cell-derived endothelial cellpodocyte |
| spellingShingle | Yoo Jin Shin Hanbi Lee Xianying Fang Sheng Cui Sun Woo Lim Kang In Lee Jae Young Lee Hong Lim Kim Yuna Oh Can Li Chul Woo Yang Gayeon You Hyeondo Lee Hyejung Mok Byung Ha Chung A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes Molecular Therapy: Nucleic Acids MT: Oligonucleotides: Therapies and Applications Fabry disease A4GALT siRNA polyhistidine-incorporating lipid nanoparticle human-induced pluripotent-stem-cell-derived endothelial cell podocyte |
| title | A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes |
| title_full | A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes |
| title_fullStr | A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes |
| title_full_unstemmed | A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes |
| title_short | A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes |
| title_sort | a4galt targeting sirna lipid nanoparticles ameliorate fabry disease phenotype greater efficacy in endothelial cells than in podocytes |
| topic | MT: Oligonucleotides: Therapies and Applications Fabry disease A4GALT siRNA polyhistidine-incorporating lipid nanoparticle human-induced pluripotent-stem-cell-derived endothelial cell podocyte |
| url | http://www.sciencedirect.com/science/article/pii/S2162253125001271 |
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