AFM nano‐mechanical study of the beating profile of hiPSC‐derived cardiomyocytes beating bodies WT and DM1

Abstract

Myotonic Dystrophy type 1 (DM1) is the most common form of muscular dystrophy in adults, characterized by a variety of multisystemic features and associated with cardiac anomalies. Among cardiac phenomena, conduction defects, ventricular arrhythmias, and dilated cardiomyopathy represent the main cause of sudden death in DM1 patients. Patient-specific induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) represent a powerful in vitro model for molecular, biochemical, and physiological studies of disease in the target cells. Here, we used an Atomic Force Microscope (AFM) to measure the beating profiles of a large number of cells, organized in CM clusters (Beating Bodies, BBs), obtained from wild type (WT) and DM1 patients. We monitored the evolution over time of the frequency and intensity of the beating. We determined the variations between different BBs and over various areas of a single BB, caused by morphological and biomechanical variations. We exploited the AFM tip to apply a controlled force over the BBs, to carefully assess the biomechanical reaction of the different cell clusters over time, both in terms of beating frequency and intensity. Our measurements demonstrated differences between the WT and DM1 clusters highlighting, for the DM1 samples, an instability which was not observed in WT cells. We measured differences in the cellular response to the applied mechanical stimulus in terms of beating synchronicity over time and cell tenacity, which are in good agreement with the cellular behavior in vivo. Overall, the combination of hiPSC-CMs with AFM characterization can become a new tool to study the collective movements of cell clusters in different conditions and can be extended to the characterization of the BB response to chemical and pharmacological stimuli.

Tools for studying and modulating (cardiac muscle) cell mechanics and mechanosensing across the scales

“…When AFM was used to measure contraction dynamics of hiPSC-derived cardiomyocytes from control and myotonic dystrophy type 1 patients, a higher mechanical resistance was observed based on altered beating impulse or beat duration (Dinarelli et al 2018). Similarly, a multi-parameter AFM-based study measured contraction force, rate and duration as well as elastic modulus in hESC-CM and iPSC-CM, allowing to map spatial heterogeneity of height, elastic modulus and contraction force, together demonstrating aberrant contractility and mechanical properties in iPSCcardiomyocytes from dilated cardiomyopathy patients (DCM) patients (Liu et al 2012).…”

Fifteen years of Servitude et Grandeur to the application of a biophysical technique in medicine: The tale of AFMBioMed

“…A virtual issue appeared in the Journal of Molecular Recognition and is linked with this editorial. Ten research articles composed this special issue Pleskova S. N., Gorshkova E. N., and Kriukov R. N. Dynamics of formation and morphological features of neutrophil extracellular traps formed under the influence of opsonized Staphylococcus aureus .…”

Aminophylline Induces Two Types of Arrhythmic Events in Human Pluripotent Stem Cell–Derived Cardiomyocytes

“…The model was originally described in previous studies (Burridge et al, 2012;Pesl et al, 2014). Its functional properties were further validated using atomic force microscopy (Liu et al, 2012;Dinarelli et al, 2018;Pribyl et al, 2019), a method enabling real-time monitoring of contractions and thus also beating properties of single cardiomyocytes or cardiac cell clusters, alone or in combination with other methods such as microelectrode array (Caluori et al, 2019b) or calcium imaging (Odstrcilik et al, 2015;Caluori et al, 2019a). This model was recently used for disease modeling in studies by Acimovic et al, 2018;Jelinkova et al, 2019. The effects of aminophylline were further validated on an independent model of HL-1 cardiomyocyte cells.…”