A human cardiomyocyte screen identifies optimized lipid nanoparticles for in vivo cardiac gene editing
Abstract
RNA therapeutics offer promising potential for treating heart disease; however, their clinical application has been limited by insufficient cardiac delivery. Here, we developed a screening platform using human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs) to identify human cardiotropic lipid nanoparticles (LNPs). By screening a chemically diverse LNP library, we identified a lead LNP, 18:1 TAP10, with potent human cardiac transfection capability. Using the Ai14 LoxP-Stop-LoxP-tdTomato reporter mouse model, we assessed 18:1 TAP10 LNP delivery of Cre mRNA in vivo via four administration routes (intravenous, intrathoracic, intracoronary, and intramyocardial (IM)) and found that each administration path enabled both the spatial distribution and cellular tropism in the heart to varying degrees. While all routes achieved robust transfection across non-CM populations (up to 40% in endothelial cells), IM injection notably enabled substantial CM transfection (36% at injection site, 13% distally). To demonstrate therapeutic potential, 18:1 TAP10 LNPs were used to deliver adenine base editor (ABE) components to Duchenne muscular dystrophy (DMD) patient-derived hiPSC-CMs, resulting in 80% on-target gene correction and restoration of dystrophin expression. In a humanized DMD mouse model, we observed dystrophin-positive cardiomyocytes in the injected left ventricle following IM administration of LNP-ABE. This work establishes a human cell-based platform for discovering organ-tropic delivery vehicles and highlights LNPs as a potential modality for therapeutic mRNA delivery and gene editing in the heart.
Article Details
Journal Info
Proceedings of the National Academy of Sciences
National Academy of Sciences
Authors (20)
Yehui Sun
Department of Biomedical Engineering, Simmons Comprehensive Cancer Center, Program in Genetic Drug Engineering, The University of Texas Southwestern Medical Center
Yu-Chung Pien
Department of Molecular Biology, The University of Texas Southwestern Medical Center
Yufen Xiao
Department of Biomedical Engineering, Simmons Comprehensive Cancer Center, Program in Genetic Drug Engineering, The University of Texas Southwestern Medical Center
Wei Tan
Efrain Sanchez-Ortiz
Department of Molecular Biology, The University of Texas Southwestern Medical Center
Mateusz Z. Durbacz
Department of Molecular Biology, The University of Texas Southwestern Medical Center
Zexiang Chen
Department of Biomedical Engineering, Simmons Comprehensive Cancer Center, Program in Genetic Drug Engineering, The University of Texas Southwestern Medical Center
John R. McAnally
Department of Molecular Biology, Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center
Yu Zhang
Xiangya Hospital, Central South University Changsha China
Hui Li
Andreas C. Chai
Department of Molecular Biology, The University of Texas Southwestern Medical Center
Francesco Chemello
Department of Molecular Biology, The University of Texas Southwestern Medical Center
Fangyu Zhang
Department of Biomedical Engineering, Simmons Comprehensive Cancer Center, Program in Genetic Drug Engineering, The University of Texas Southwestern Medical Center
Sang M. Lee
Department of Biomedical Engineering, Simmons Comprehensive Cancer Center, Program in Genetic Drug Engineering, The University of Texas Southwestern Medical Center
Priyanka Patel
Department of Biomedical Engineering, Simmons Comprehensive Cancer Center, Program in Genetic Drug Engineering, The University of Texas Southwestern Medical Center
Denise M. Ramirez
Department of Neurology, The University of Texas Southwestern Medical Center
Sumanta Chatterjee
Department of Biomedical Engineering, Simmons Comprehensive Cancer Center, Program in Genetic Drug Engineering, The University of Texas Southwestern Medical Center
Ning Liu
Eric N. Olson
Department of Molecular Biology, Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center
Daniel J. Siegwart
Department of Biomedical Engineering, Simmons Comprehensive Cancer Center, Program in Genetic Drug Engineering, The University of Texas Southwestern Medical Center