Long-term culture of patient-derived cardiac organoids recapitulated Duchenne muscular dystrophy cardiomyopathy and disease progression

Vittoria Marini; Fabiola Marino; Flaminia Aliberti; Nefele Giarratana; Enrico Pozzo; Robin Duelen; Álvaro Cortés Calabuig; Rita La Rovere; Tim Vervliet; Daniele Torella; Geert Bultynck; Maurilio Sampaolesi; Yoke Chin Chai

doi:10.3389/fcell.2022.878311

Long-term culture of patient-derived cardiac organoids recapitulated Duchenne muscular dystrophy cardiomyopathy and disease progression

Front Cell Dev Biol. 2022 Aug 11:10:878311. doi: 10.3389/fcell.2022.878311. eCollection 2022.

Authors

Affiliations

¹ Translational Cardiomyology Laboratory, Stem Cell Biology and Embryology, Department of Development and Regeneration, KU Leuven, Leuven, Belgium.
² Department of Experimental and Clinical Medicine, Magna Graecia University, Catanzaro, Italy.
³ Fondazione IRCCS Policlinico San Matteo, Center for Inherited Cardiovascular Diseases, Transplant Research Area, Human Anatomy Unit, Department of Public Health, Experimental and Forensic Medicine, University of Pavia, Pavia, Italy.
⁴ Genomics Core Leuven, KU Leuven, Leuven, Belgium.
⁵ Laboratory of Molecular and Cellular Signaling, Department of Cellular and Molecular Medicine and Leuven Kanker Institute, KU Leuven, Leuven, Belgium.
⁶ Histology and Medical Embryology Unit, Department of Anatomy, Histology, Forensic Medicine and Orthopedics, Sapienza University of Rome, Rome, Italy.

Abstract

Duchenne Muscular Dystrophy (DMD) is an X-linked neuromuscular disease which to date is incurable. The major cause of death is dilated cardiomyopathy however, its pathogenesis is unclear as existing cellular and animal models do not fully recapitulate the human disease phenotypes. In this study, we generated cardiac organoids from patient-derived induced pluripotent stem cells (DMD-COs) and isogenic-corrected controls (DMD-Iso-COs) and studied if DMD-related cardiomyopathy and disease progression occur in the organoids upon long-term culture (up to 93 days). Histological analysis showed that DMD-COs lack initial proliferative capacity, displayed a progressive loss of sarcoglycan localization and high stress in endoplasmic reticulum. Additionally, cardiomyocyte deterioration, fibrosis and aberrant adipogenesis were observed in DMD-COs over time. RNA sequencing analysis confirmed a distinct transcriptomic profile in DMD-COs which was associated with functional enrichment in hypertrophy/dilated cardiomyopathy, arrhythmia, adipogenesis and fibrosis pathways. Moreover, five miRNAs were identified to be crucial in this dysregulated gene network. In conclusion, we generated patient-derived cardiac organoid model that displayed DMD-related cardiomyopathy and disease progression phenotypes in long-term culture. We envision the feasibility to develop a more complex, realistic and reliable in vitro 3D human cardiac-mimics to study DMD-related cardiomyopathies.

Keywords: Duchenne muscular dystrophy; aberrant adipogenesis; cardiac organoids; cardiomyopathy; disease modeling; fibrosis; induced pluripotent stem cells.