A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes

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

doi:10.1016/j.omtn.2025.102573

A4GALT-targeting siRNA lipid nanoparticles ameliorate Fabry disease phenotype: Greater efficacy in endothelial cells than in podocytes

Mol Ther Nucleic Acids. 2025 May 20;36(2):102573. doi: 10.1016/j.omtn.2025.102573. eCollection 2025 Jun 10.

Authors

Yoo Jin Shin¹, Hanbi Lee^{1

2}, Xianying Fang¹, Sheng Cui¹, Sun Woo Lim¹, Kang In Lee³, Jae Young Lee^{3

4}, Hong Lim Kim⁵, Yuna Oh⁵, Can Li⁶, Chul Woo Yang^{1

2}, Gayeon You⁷, Hyeondo Lee⁷, Hyejung Mok⁷, Byung Ha Chung^{1

2}

Affiliations

¹ Transplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.
² 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.
³ R&D Center, ToolGen, Inc., Seoul 07789, Republic of Korea.
⁴ Department of Anatomy, Ajou University School of Medicine, Suwon 16499, Korea.
⁵ Integrative Research Support Center, Laboratory of Electron Microscope, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.
⁶ Department of Nephrology, Yanbian University Hospital, Yanji, China.
⁷ Department of Bioscience and Biotechnology, Konkuk University, Seoul 05029, Republic of Korea.

Abstract

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.

Keywords: A4GALT siRNA; Fabry disease; MT: Oligonucleotides: Therapies and Applications; human-induced pluripotent-stem-cell-derived endothelial cell; podocyte; polyhistidine-incorporating lipid nanoparticle.