DMD Pluripotent Stem Cell Derived Cardiac Cells Recapitulate in vitro Human Cardiac Pathophysiology

Sarka Jelinkova; Aleksandra Vilotic; Jan Pribyl; Franck Aimond; Anton Salykin; Ivana Acimovic; Martin Pesl; Guido Caluori; Simon Klimovic; Tomas Urban; Hana Dobrovolna; Vladimir Soska; Petr Skladal; Alain Lacampagne; Petr Dvorak; Albano C Meli; Vladimir Rotrekl

doi:10.3389/fbioe.2020.00535

DMD Pluripotent Stem Cell Derived Cardiac Cells Recapitulate in vitro Human Cardiac Pathophysiology

Front Bioeng Biotechnol. 2020 Jun 19:8:535. doi: 10.3389/fbioe.2020.00535. eCollection 2020.

Authors

Sarka Jelinkova^{1

2}, Aleksandra Vilotic¹, Jan Pribyl³, Franck Aimond⁴, Anton Salykin¹, Ivana Acimovic¹, Martin Pesl^{1

2

5}, Guido Caluori^{2

5}, Simon Klimovic⁶, Tomas Urban¹, Hana Dobrovolna⁷, Vladimir Soska^{7

8}, Petr Skladal^{5

6}, Alain Lacampagne⁴, Petr Dvorak^{1

2}, Albano C Meli^{1

4}, Vladimir Rotrekl^{1

2}

Affiliations

¹ Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czechia.
² International Clinical Research Center ICRC, St. Anne's University Hospital Brno, Brno, Czechia.
³ CEITEC, Masaryk University, Brno, Czechia.
⁴ PhyMedExp, University of Montpellier, INSERM, CNRS, Montpellier, France.
⁵ First Department of Internal Medicine-Cardioangiology, Faculty of Medicine, Masaryk University, Brno, Czechia.
⁶ Department of Biochemistry, Faculty of Science, Masaryk University, Brno, Czechia.
⁷ Department of Clinical Biochemistry, St. Anne's University Hospital of Brno, Brno, Czechia.
⁸ Second Clinic of Internal Medicine, Masaryk University of Brno, Brno, Czechia.

Abstract

Duchenne muscular dystrophy (DMD) is a severe genetic disorder characterized by the lack of functional dystrophin. DMD is associated with progressive dilated cardiomyopathy, eventually leading to heart failure as the main cause of death in DMD patients. Although several molecular mechanisms leading to the DMD cardiomyocyte (DMD-CM) death were described, mostly in mouse model, no suitable human CM model was until recently available together with proper clarification of the DMD-CM phenotype and delay in cardiac symptoms manifestation. We obtained several independent dystrophin-deficient human pluripotent stem cell (hPSC) lines from DMD patients and CRISPR/Cas9-generated DMD gene mutation. We differentiated DMD-hPSC into cardiac cells (CC) creating a human DMD-CC disease model. We observed that mutation-carrying cells were less prone to differentiate into CCs. DMD-CCs demonstrated an enhanced cell death rate in time. Furthermore, ion channel expression was altered in terms of potassium (Kir2.1 overexpression) and calcium handling (dihydropyridine receptor overexpression). DMD-CCs exhibited increased time of calcium transient rising compared to aged-matched control, suggesting mishandling of calcium release. We observed mechanical impairment (hypocontractility), bradycardia, increased heart rate variability, and blunted β-adrenergic response connected with remodeling of β-adrenergic receptors expression in DMD-CCs. Overall, these results indicated that our DMD-CC models are functionally affected by dystrophin-deficiency associated and recapitulate functional defects and cardiac wasting observed in the disease. It offers an accurate tool to study human cardiomyopathy progression and test therapies in vitro.

Keywords: DMD; adrenergic response; cardiomyocyte death; cardiomyocytes; duchenne muscular dystrophy; excitation-contraction coupling; human pluripotent stem cells; intracellular calcium.