Long-Term Culture of Patient-Derived Cardiac Organoids Recapitulated Duchenne Muscular Dystrophy Cardiomyopathy and Disease Progression

Abstract

Duchenne Muscular Dystrophy (DMD) is an X-linked neuromuscular disease which to-date incurable. The major cause of death is dilated cardiomyopathy, however the 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 pluripotent stem cells (DMD-CO) and isogenic-corrected controls (DMD-Iso-CO) 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-CO lacks initial proliferative capacity, displayed a progressive loss of α-sarcoglycan localization and high stress in endoplasmic reticulum. Additionally, the cardiomyocyte deteriorated over time, and fibrosis and adipogenesis were observed in DMD-CO. RNA sequencing analysis confirmed a distinct transcriptomic profile in DMD-CO which were 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.

From bedside to the bench: patient-specific hiPSC-EC models uncover endothelial dysfunction in genetic cardiomyopathies

“…This is known to increase CM maturation, possibly due to cardiac endothelium cardiotropic function (Segers et al, 2018), and to induce vascularization. Only few studies, in which hiPSC-EC were included in a MT or EHT, have modelled cardiomyopathies, such as ACM (Giacomelli et al, 2020), LQTS (Giacomelli et al, 2021), and Duchenne muscular dystrophyassociated cardiomyopathy (Marini et al, 2022), with no specific reference to ED and its effect on the tissue in vitro. In addition, most cardiac organoids, obtained by differentiation and self-organization of hiPSCs show a proportion of ECs (Mills et al, 2017;Lewis-Israeli et al, 2021).…”

Extracellular vesicles and Duchenne muscular dystrophy pathology: Modulators of disease progression

“…More recently, it was discovered that exosomes isolated from DMD muscle-derived fibroblasts stimulate fibrosis, mainly mediated by exosomal miR-199-5p (Zanotti et al, 2018). With regard to three dimensional (3D) cardiac models, recently, our laboratory unveiled that cardiac organoids derived from DMD iPSCs display DMD-like cardiomyopathy and disease progression phenotypes in long-term cultures compared to clustered regularly interspaced short palindromic repeats (CRISPR)/(CRISPR/Cas9) mutationcorrected isogenic controls (Marini et al, 2022). It was revealed that in the DMD cardiac organoid dysregulated gene network, three miRNAs were found to play crucial roles.…”

Exploring the prospects, advancements, and challenges of in vitro modeling of the heart-brain axis

“…Assembloids have recapitulated cortico-thalamic, cortical-subpallium, cortico-striatal, gut-brain, visual system, and hindbrain-skeletal muscle interactions (Bagley et al, 2017;Birey et al, 2017;Xiang et al, 2019;Andersen et al, 2020;Miura et al, 2020;Fligor et al, 2021;Reiner et al, 2021). Morphogens guide cell fate differentiation, enhance reproducibility, and yield brain organoids with specific cell fates (Muguruma et al, 2015;Birey et al, 2017;Xiang et al, 2017Xiang et al, , 2019 and cardiac organoids with neurons or endothelial cells resembling fetal heart development (Marini et al, 2022). Culturing adult cell types is difficult, but in vitro modeling of Huntington's and Parkinson's has identified early markers of these midlife or late-life diseases (Smits et al, 2019;Andrews and Kriegstein, 2022).…”