Ibrahim El-Battrawy1, Zhihan Zhao1, Huan Lan1, Xin Li1, Gökhan Yücel1, Siegfried Lang1, Katherine Sattler1, Jan-Dierk Schünemann1, Wolfram-Hubertus Zimmermann1, Lukas Cyganek1, Jochen Utikal1, Thomas Wieland1, Karen Bieback1, Ralf Bauer1, Antonius Ratte1, Regina Pribe-Wolferts1, Kleopatra Rapti1, Daniel Nowak1, Janina Wittig1, Dierk Thomas1, Patrick Most1, Hugo A Katus1, Ursula Ravens1, Constanze Schmidt1, Martin Borggrefe1, Xiao-Bo Zhou2, Oliver J Müller1, Ibrahim Akin1. 1. From the First Department of Medicine, Faculty of Medicine (I.E.-B., Z.Z., H.L., X.L., G.Y., S.L., K.S., J.-D.S., M.B., X.-B.Z., I.A.) and Department of Dermatology, Venereology and Allergology (J.U.), University Medical Centre Mannheim, University of Heidelberg, Germany; DZHK (German Center for Cardiovascular Research), Partner Sites, Heidelberg-Mannheim and Göttingen (I.E.-B., Z.Z., H.L., G.Y., S.L., W.-H.Z., L.C., J.U., T.W., R.B., A.R., D.T., P.M., H.A.K., C.S., M.B., X.-B.Z., O.J.M., I.A.); Institute of Pharmacology and Toxicology, University of Göttingen, Germany (W.-H.Z.); Stem Cell Unit, Clinic for Cardiology and Pneumology, University Medical Center Göttingen, Germany (L.C.); Skin Cancer Unit, German Cancer Research Center (DKFZ), Heidelberg (J.U.); Institute of Experimental and Clinical Pharmacology and Toxicology (T.W.) and Department of Hematology and Oncology (D.N., J.W.), Medical Faculty Mannheim, University of Heidelberg, Germany; Institute for Transfusion Medicine and Immunology, Mannheim, Germany (K.B.); Internal Medicine III, University Hospital Heidelberg, Germany (R.B., A.R., R.P.-W., K.R., D.T., P.M., H.A.K., C.S., O.J.M.); Institute of Experimental Cardiovascular Medicine, University Heart Centre Freiburg, Germany (U.R.); Medical Faculty, University of Freiburg, Germany (U.R.); and Key Laboratory of Medical Electrophysiology of Ministry of Education, Institute of Cardiovascular Research, Southwest Medical University, Luzhou, China (H.L., X.-B.Z.). 2. From the First Department of Medicine, Faculty of Medicine (I.E.-B., Z.Z., H.L., X.L., G.Y., S.L., K.S., J.-D.S., M.B., X.-B.Z., I.A.) and Department of Dermatology, Venereology and Allergology (J.U.), University Medical Centre Mannheim, University of Heidelberg, Germany; DZHK (German Center for Cardiovascular Research), Partner Sites, Heidelberg-Mannheim and Göttingen (I.E.-B., Z.Z., H.L., G.Y., S.L., W.-H.Z., L.C., J.U., T.W., R.B., A.R., D.T., P.M., H.A.K., C.S., M.B., X.-B.Z., O.J.M., I.A.); Institute of Pharmacology and Toxicology, University of Göttingen, Germany (W.-H.Z.); Stem Cell Unit, Clinic for Cardiology and Pneumology, University Medical Center Göttingen, Germany (L.C.); Skin Cancer Unit, German Cancer Research Center (DKFZ), Heidelberg (J.U.); Institute of Experimental and Clinical Pharmacology and Toxicology (T.W.) and Department of Hematology and Oncology (D.N., J.W.), Medical Faculty Mannheim, University of Heidelberg, Germany; Institute for Transfusion Medicine and Immunology, Mannheim, Germany (K.B.); Internal Medicine III, University Hospital Heidelberg, Germany (R.B., A.R., R.P.-W., K.R., D.T., P.M., H.A.K., C.S., O.J.M.); Institute of Experimental Cardiovascular Medicine, University Heart Centre Freiburg, Germany (U.R.); Medical Faculty, University of Freiburg, Germany (U.R.); and Key Laboratory of Medical Electrophysiology of Ministry of Education, Institute of Cardiovascular Research, Southwest Medical University, Luzhou, China (H.L., X.-B.Z.). Xiaobo.zhou@medma.uni-heidelberg.de.
Abstract
BACKGROUND: Limb-Girdle muscular dystrophies (LGMD) are a heritable group of genetically determined disorders with a primary involvement of the pelvic or shoulder girdle musculature with partially cardiac manifestation, such as dilated cardiomyopathy (DCM) and life-threatening tachyarrhythmia. We report here that human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes from a patient with LGMD2I and DCM associated with recurrent ventricular tachycardia displayed ion channel dysfunction and abnormality of calcium homeostasis. METHODS: Dermal fibroblasts obtained from a patient with LGMD2I harboring a fukutin-related protein gene mutation (826C>A; Leu276Ile) and 3 healthy donors were reprogrammed to hiPSCs. The hiPSCs were differentiated into cardiomyocytes and used for biological and electrophysiological studies. RESULTS: Compared with hiPSC cardiomyocytes from the healthy donors, the hiPSC cardiomyocytes from the patient exhibited abnormal action potentials characterized by reduced amplitude and upstroke velocity. The peak and late Na channel currents (INa) as well as the peak L-type calcium channel currents were significantly reduced. The expression of SCN5A and CACNA1C was reduced in DCM cardiomyocytes, consistent with reduction of INa and L-type calcium channel currents. In addition, the rapidly activating delayed rectifier potassium current (IKr) was reduced, whereas the transient outward current (Ito) and slowly activating delayed rectifier potassium current (IKs) were similar in DCM and control cardiomyocytes. Finally, a significant reduction of systolic and diastolic intracellular Ca2+ concentrations was detected in DCM cardiomyocytes. CONCLUSIONS: This study demonstrates that patient-specific hiPSC cardiomyocytes can recapitulate some phenotypic properties of LGMD2I with DCM and provide a platform for studies on the cardiac events in LGMD.
BACKGROUND:Limb-Girdle muscular dystrophies (LGMD) are a heritable group of genetically determined disorders with a primary involvement of the pelvic or shoulder girdle musculature with partially cardiac manifestation, such as dilated cardiomyopathy (DCM) and life-threatening tachyarrhythmia. We report here that human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes from a patient with LGMD2I and DCM associated with recurrent ventricular tachycardia displayed ion channel dysfunction and abnormality of calcium homeostasis. METHODS: Dermal fibroblasts obtained from a patient with LGMD2I harboring a fukutin-related protein gene mutation (826C>A; Leu276Ile) and 3 healthy donors were reprogrammed to hiPSCs. The hiPSCs were differentiated into cardiomyocytes and used for biological and electrophysiological studies. RESULTS: Compared with hiPSC cardiomyocytes from the healthy donors, the hiPSC cardiomyocytes from the patient exhibited abnormal action potentials characterized by reduced amplitude and upstroke velocity. The peak and late Na channel currents (INa) as well as the peak L-type calcium channel currents were significantly reduced. The expression of SCN5A and CACNA1C was reduced in DCM cardiomyocytes, consistent with reduction of INa and L-type calcium channel currents. In addition, the rapidly activating delayed rectifier potassium current (IKr) was reduced, whereas the transient outward current (Ito) and slowly activating delayed rectifier potassium current (IKs) were similar in DCM and control cardiomyocytes. Finally, a significant reduction of systolic and diastolic intracellular Ca2+ concentrations was detected in DCM cardiomyocytes. CONCLUSIONS: This study demonstrates that patient-specific hiPSC cardiomyocytes can recapitulate some phenotypic properties of LGMD2I with DCM and provide a platform for studies on the cardiac events in LGMD.
Authors: Andreas Brodehl; Hans Ebbinghaus; Marcus-André Deutsch; Jan Gummert; Anna Gärtner; Sandra Ratnavadivel; Hendrik Milting Journal: Int J Mol Sci Date: 2019-09-06 Impact factor: 5.923