PMC7494730_01
Contractile properties of LMNA-mutant DCM hiPSCMs assessed in the 2D sheet approach. (A) (Left) Pedigree of a familial DCM cohort with a heterozygous mutation in LMNA. Black arrow indicates the DCM patient; white arrow indicates the healthy sibling who served as a healthy control (HC). (Right) Sanger sequencing of DNA from the patient indicating a heterozygous mutation in LMNA. (B) (Left) Representative images of impedance recording obtained from xCELLigence CardioECR. Red, recording from the HC sample; green, recording from the DCM patient sample. Amp, amplitude. (Upper Right) Comparison of beating rate and impedance amplitude obtained from HC and DCM hiPSCM sheets (HC, n = 12; DCM, n = 13). CI, cellular impedance. (Lower Right) Comparison of beating rate and impedance amplitude of HC hiPSCM sheets before and after treatment with 100 nM isoproterenol (n = 10, respectively). (C) (Left) Conceptual image of the Cell Motion Imaging System and parameters calculated by the device. CV, contraction velocity; RV, relaxation velocity. (Upper Right) Comparison of beating rate, CV, and RV obtained from HC and DCM hiPSCM sheets (HC, n = 7; DCM, n = 6). (Upper Left) Comparison of beating rate, CV, and RV of HC hiPSCM sheets before and after treatment with 100 nM isoproterenol (n = 8, respectively). *p < 0.05; **p < 0.01; ***p < 0.001.
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F1
A heterozygous missense mutation in LMNA (p.Q353R) was revealed by panel sequencing of candidate cardiomyopathy genes (Figure 1A). hiPSCMs were produced from a male patient with an LMNA mutation and his healthy sibling (Figure 1A, Supplementary Figure 1). These results suggest that this system partially reproduced the functional abnormalities of DCM; however, it would not be suitable for analysis of the pharmacological response including beta-adrenergic receptor signaling (Figure 1B, Supplementary Figure 3). On the other hand, when isoproterenol was added to the hiPSCMs, CV and RV were both increased significantly, which suggested that the positive inotropic and lusitropic effects of the beta-adrenergic stimulant could be assessed using this system (Figure 1C).