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| Status |
Public on May 31, 2024 |
| Title |
PITX2 knockout induces key findings of electrical remodeling as seen in persistent atrial fibrillation |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Introduction: Atrial fibrillation (AF) is the most common arrhythmia and affects around 1% of the population with increasing incidence and substantial morbidity. Functional effects of genetic variants associated with AF should help to identify targets involved in AF initiation and/or subsequent remodeling. Gene variants next to the gene of the transcription factor PITX2 (paired-like homeodomain transcription factor 2) showed the strongest association with AF. A significant number of AF patients showed reduced PITX2 levels in the left atrium. Mouse models have been used to study functional aspects of PITX2 gene deletion (“knock out”), but there are substantial differences in atrial electrophysiology between mouse and human . HiPSC-CM should be able to bridge the gap, provided the cells reproduce typical characteristics of human atrium. Methods: PITX2 knock out samples from a control cell line of human induced pluripotent stem cells (hiPSC) were obtained. Differentiation and generation of Atrial-like engineered heart tissue (aEHT) was performed. RNA-sequencing (RNA-Seq), Quantitative Real-time PCR (RT-qPCR), Protein analysis by Western Blot, Contraction and force analysis, action potential measurements, Calcium current measurements and differential gene expression analysis were run on the samples. Results: Effective knock out of PITX2 was confirmed by mRNA sequencing and WB. It was shown that PITX2 deficiency reduces contraction force. PITX2 deficiency induces action potential triangulation with more negative maximum diastolic potential. Activation of muscarinic receptors shortens APD in both PITX2 knockout and controls but hyperpolarization is lost in PITX2 knock out. It was shown that Ca2+ current density is smaller in PITX2 knock out than controls. We found that more negative MDP in PITX2 knock out is not associated with higher IK1. Conclusion: We improved atrial differentiation, coming very close to human atrial electrophysiology. We demonstrate that PITX2 deficiency induces key characteristics known from persistent AF.
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| Overall design |
Control group (Cell line of human induced pluripotent stem cells): 3 samples; Study group (PITX2 Knockout): 3 samples
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| Contributor(s) |
Schulz C, Lemoine MD, Mearini G, Sani J, Koivumäki J, Ghalawinji A, Stoll M, Kirchhof P, Hansen A, Eschenhagen T, Christ T |
| Citation missing |
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| |
| Submission date |
Jun 01, 2021 |
| Last update date |
May 31, 2024 |
| Contact name |
Amer Ghalawinji |
| E-mail(s) |
amer.ghalawinji@uni-muenster.de
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| Organization name |
WWU
|
| Department |
Institute of Human Genetics
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| Lab |
Department of Genetic Epidemiology
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| Street address |
Domagkstrasse 3
|
| City |
Muenster |
| ZIP/Postal code |
48149 |
| Country |
Germany |
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| Platforms (1) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA734337 |
| SRA |
SRP322186 |
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