scholarly journals Cardiac microtissues from human pluripotent stem cells recapitulate the phenotype of long-QT syndrome

2021 ◽  
Vol 572 ◽  
pp. 118-124
Author(s):  
Elisa Giacomelli ◽  
Luca Sala ◽  
Dorien Ward-van Oostwaard ◽  
Milena Bellin
Keyword(s):  
Stem Cells ◽  
Long Qt Syndrome ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Long Qt ◽  
Qt Syndrome

scholarly journals Base Editing of Human Pluripotent Stem Cells for modeling Long QT syndrome

2021 ◽  
Author(s):  
Fujian Wu ◽  
Tianwei Guo ◽  
Lixiang Sun ◽  
Furong Li ◽  
Xiaofei Yang
Keyword(s):  
Stem Cells ◽  
Long Qt Syndrome ◽  
Pluripotent Stem Cells ◽  
Disease Modeling ◽  
Human Pluripotent Stem Cells ◽  
Electrode Arrays ◽  
Long Qt ◽  
Base Editing ◽  
Qt Syndrome ◽  
Disease Specific

Abstract Human pluripotent stem cells (hPSCs) have great potential for disease modeling, drug discovery, and regenerative medicine as they can differentiate into many different functional cell types via directed differentiation. However, the application of disease modeling is limited due to a time-consuming and labor-intensive process of introducing known pathogenic mutations into hPSCs. Base editing is a newly developed technology that enables the facile introduction of point mutations into specific loci within the genome of living cells without unwanted genome injured. We describe an optimized stepwise protocol to introduce disease-specific mutations of long QT syndrome (LQTs) into hPSCs. We highlight technical issues, especially those associated with introducing a point mutation to obtain isogenic hPSCs without inserting any resistance cassette and reproducible cardiomyocyte differentiation. Based on the protocol, we succeeded in getting hPSCs carrying LQTs pathogenic mutation with excellent efficiency (21.7 % of heterozygous clones, 9.1 % of homozygous clones) in less than 20 days. In addition, we also provide protocols to analyze electrophysiological of hPSC-derived cardiomyocytes using multi-electrode arrays. This protocol is also applicable to introduce other disease-specific mutations into hPSCs.

scholarly journals Base Editing of Human Pluripotent Stem Cells for Modeling Long QT Syndrome

2022 ◽  
Author(s):  
Fujian Wu ◽  
Tianwei Guo ◽  
Lixiang Sun ◽  
Furong Li ◽  
Xiaofei Yang
Keyword(s):  
Stem Cells ◽  
Long Qt Syndrome ◽  
Pluripotent Stem Cells ◽  
Disease Modeling ◽  
Human Pluripotent Stem Cells ◽  
Electrode Arrays ◽  
Long Qt ◽  
Base Editing ◽  
Qt Syndrome ◽  
Disease Specific

AbstractHuman pluripotent stem cells (hPSCs) have great potential for disease modeling, drug discovery, and regenerative medicine as they can differentiate into many different functional cell types via directed differentiation. However, the application of disease modeling is limited due to a time-consuming and labor-intensive process of introducing known pathogenic mutations into hPSCs. Base editing is a newly developed technology that enables the facile introduction of point mutations into specific loci within the genome of living cells without unwanted genome injured. We describe an optimized stepwise protocol to introduce disease-specific mutations of long QT syndrome (LQTs) into hPSCs. We highlight technical issues, especially those associated with introducing a point mutation to obtain isogenic hPSCs without inserting any resistance cassette and reproducible cardiomyocyte differentiation. Based on the protocol, we succeeded in getting hPSCs carrying LQTs pathogenic mutation with excellent efficiency (31.7% of heterozygous clones, 9.1% of homozygous clones) in less than 20 days. In addition, we also provide protocols to analyze electrophysiological of hPSC-derived cardiomyocytes using multi-electrode arrays. This protocol is also applicable to introduce other disease-specific mutations into hPSCs. Graphical abstract

scholarly journals From patient-specific induced pluripotent stem cells to clinical translation in long QT syndrome Type 2

2019 ◽  
Vol 40 (23) ◽  
pp. 1832-1836 ◽  
Author(s):  
Peter J Schwartz ◽  
Massimiliano Gnecchi ◽  
Federica Dagradi ◽  
Silvia Castelletti ◽  
Gianfranco Parati ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Long Qt Syndrome ◽  
Pluripotent Stem Cells ◽  
Patient Specific ◽  
Syndrome Type ◽  
Long Qt ◽  
Qt Syndrome ◽  
Induced Pluripotent

scholarly journals Induced pluripotent stem cells used to reveal drug actions in a long QT syndrome family with complex genetics

2012 ◽  
Vol 141 (1) ◽  
pp. 61-72 ◽  
Author(s):  
Cecile Terrenoire ◽  
Kai Wang ◽  
Kelvin W. Chan Tung ◽  
Wendy K. Chung ◽  
Robert H. Pass ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Long Qt Syndrome ◽  
Pluripotent Stem Cells ◽  
Na Channel ◽  
Long Qt ◽  
Complex Genetics ◽  
Drug Actions ◽  
Qt Syndrome ◽  
Induced Pluripotent

Understanding the basis for differential responses to drug therapies remains a challenge despite advances in genetics and genomics. Induced pluripotent stem cells (iPSCs) offer an unprecedented opportunity to investigate the pharmacology of disease processes in therapeutically and genetically relevant primary cell types in vitro and to interweave clinical and basic molecular data. We report here the derivation of iPSCs from a long QT syndrome patient with complex genetics. The proband was found to have a de novo SCN5A LQT-3 mutation (F1473C) and a polymorphism (K897T) in KCNH2, the gene for LQT-2. Analysis of the biophysics and molecular pharmacology of ion channels expressed in cardiomyocytes (CMs) differentiated from these iPSCs (iPSC-CMs) demonstrates a primary LQT-3 (Na+ channel) defect responsible for the arrhythmias not influenced by the KCNH2 polymorphism. The F1473C mutation occurs in the channel inactivation gate and enhances late Na+ channel current (INaL) that is carried by channels that fail to inactivate completely and conduct increased inward current during prolonged depolarization, resulting in delayed repolarization, a prolonged QT interval, and increased risk of fatal arrhythmia. We find a very pronounced rate dependence of INaL such that increasing the pacing rate markedly reduces INaL and, in addition, increases its inhibition by the Na+ channel blocker mexiletine. These rate-dependent properties and drug interactions, unique to the proband’s iPSC-CMs, correlate with improved management of arrhythmias in the patient and provide support for this approach in developing patient-specific clinical regimens.

Modelling the long QT syndrome with induced pluripotent stem cells

Nature ◽  
2011 ◽  
Vol 471 (7337) ◽  
pp. 225-229 ◽  
Author(s):  
Ilanit Itzhaki ◽  
Leonid Maizels ◽  
Irit Huber ◽  
Limor Zwi-Dantsis ◽  
Oren Caspi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Long Qt Syndrome ◽  
Pluripotent Stem Cells ◽  
Long Qt ◽  
Qt Syndrome ◽  
Induced Pluripotent
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