ORIGINAL RESEARCH article
Front. Immunol.
Sec. Alloimmunity and Transplantation
GJA5 and ATP1A1 perturbations recapitulate inflammation-related beat irregularities in iPSC-based atrial myocardium tissue model
Thomas Hutschalik 1,2
Albert Dasí 3
Leto L Riebel 3
Maury Wiendels 4
Frederikus Bakker 5
Lucas J.A.M. Beckers 5
Koen C. Kriege 5
Susanne M. Valster 5
Roland C.M. Vulders 5
Ozan Özgül 1
Rémi Peyronnet 6
Blanca Rodriguez 3
Mariana Argenziano 2
Ulrich Schotten 1,7
Elena Matsa 8,2,9
1. Universiteit Maastricht Cardiovascular Research Institute Maastricht, Maastricht, Netherlands
2. Ncardia Netherlands, Leiden, Netherlands
3. University of Oxford, Oxford, United Kingdom
4. Leids Universitair Medisch Centrum, Leiden, Netherlands
5. Koninklijke Philips NV, Eindhoven, Netherlands
6. Universitats-Herzzentrum Freiburg Bad Krozingen Institut fur Experimentelle Kardiovaskulare Medizin, Freiburg, Germany
7. Maastricht Universitair Medisch Centrum+, Maastricht, Netherlands
8. University College Cork, Cork, Ireland
9. National Institute for Bioprocessing Research and Training, Dublin, Ireland
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Abstract
Atrial fibrillation (AF) is the most common cardiac arrhythmia, linked to greater risk of heart failure, stroke and death. Inflammation has been connected to AF emergence, however mechanisms of inflammation-caused AF remain thus far elusive, leading to a lack of mechanism-based treatments. An isogenic, 3D tissue model containing hiPSC-derived atrial-like cardiomyocytes (aCM), cardiac fibroblasts (cfb), and cardiac macrophages was engineered using custom injection-molded pillar devices. Electrophysiological changes were examined via sharp electrode recordings, calcium imaging, and multi-electrode assays. Gene function was interrogated using siRNA knock-down, lentiviral overexpression, and pharmacological modulation. In silico tissue and whole-heart models validated findings under simulated stress and heterogeneous conditions. Activation of M1 macrophages led to a 50% reduction in contraction amplitude, action potential spike amplitude (aCM+cfb+M1: 61.3 mV ±13.9 vs control: 71.6 mV ±14.5, p < 0.01) and increased beat irregularity (M1: 150.7% ±388.9 vs control, p < 0.001). Calcium transient amplitude was reduced (12.3 a.u. ±14.7, p < 0.05) and upstroke velocity slowed. SCN5A knock-down reduced contraction amplitude (−51.9% ±37.2, p < 0.01) without inducing arrhythmias, whereas combined GJA5 and ATP1A1 knockdown induced significant irregularity (403% ±371.3, p < 0.001), increased conduction heterogeneity (+18%), and reduced velocity (−52.4%). In silico modeling confirmed that paired 50% downregulation of sodium-potassium pump and tissue conductivity induced AF under tachycardia even without ectopic activity. This work reveals a novel, inflammation-driven mechanism for AF initiation. Combined downregulation of GJA5 (connexin 40) and ATP1A1 (NaK ATPase) disrupted intercellular connectivity and ion flux, establishing a substrate for arrhythmogenesis. These results were robust across in vitro, genetic/pharmacological, and in silico models, defining new avenues for translational intervention.
Summary
Keywords
Atrial Fibrillation, Engineered 3D tissue, Inflammation, IPSC, macrophage
Received
06 October 2025
Accepted
05 December 2025
Copyright
© 2025 Hutschalik, Dasí, Riebel, Wiendels, Bakker, Beckers, Kriege, Valster, Vulders, Özgül, Peyronnet, Rodriguez, Argenziano, Schotten and Matsa. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Elena Matsa
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