Analysis of calcium transients in cardiomyocytes derived from
hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy
controls

a 2024

Analysis of calcium transients in cardiomyocytes derived from hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy controls

ŠVECOVÁ, Olga, Martin KRÁL, Štefan ZELENÁK, Jiří PACHERNÍK, Tomáš BÁRTA et. al.

Základní údaje

Originální název

Analysis of calcium transients in cardiomyocytes derived from hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy controls

Autoři

ŠVECOVÁ, Olga (203 Česká republika, domácí), Martin KRÁL (203 Česká republika, domácí), Štefan ZELENÁK (703 Slovensko), Jiří PACHERNÍK (203 Česká republika), Tomáš BÁRTA (203 Česká republika, domácí), Samuel LIETAVA (703 Slovensko, domácí), Iva SYNKOVÁ (203 Česká republika, domácí), Jana ZÍDKOVÁ (203 Česká republika, domácí), Tomáš NOVOTNÝ (203 Česká republika, domácí) a Markéta BÉBAROVÁ (203 Česká republika, garant, domácí)

Vydání

15th New Frontiers in Basic Cardiovascular Research: A France-New EU Members Symposium, 2024

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

30201 Cardiac and Cardiovascular systems

Stát vydavatele

Francie

Utajení

není předmětem státního či obchodního tajemství

Organizační jednotka

Lékařská fakulta

Klíčová slova anglicky

calcium transient; hiPSC-derived cardiomyocytes

Příznaky

Mezinárodní význam

Změněno: 19. 8. 2024 14:13, Mgr. Tereza Miškechová

Anotace

V originále

Introduction: the regulation of intracellular calcium levels is crucial for excitation-contraction coupling. The release of calcium into the intracellular space is controlled by the ryanodine receptor type 2 (RYR2) located on the sarcoplasmic reticulum. Dysfunction of RYR2 is involved in the pathogenesis of inherited and non-inherited diseases such as cardiac arrhythmias, ventricular fibrillation, ventricular tachycardia, etc. The variant p. Y4734C in RYR2 was found in a patient with idiopathic ventricular fibrillation without structural changes in the heart and signs of arrhythmia on clinical examination. Preliminary data show calcium transient of patient-specific cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM). Methods: calcium transients of hiPSC-CM (Y4734C) and hiPSC-CM unrelated healthy controls (WT) were measured using the Myocyte Calcium and Contractility System (IonOptix LLC). Cell clusters were loaded with Fura-2 (Molecular Probes, Invitrogen) at a final concentration of 1 mM. Cells were incubated for 15 min in Tyrode solution with 1 μmol/L Fura-2-am at 37 °C and then washed repeatedly with Tyrode solution followed by incubation for 10 min in Tyrode solution at 37 °C. Measurements were performed in Tyrode solution at 37 ± 0.5 °C. Cells were not stimulated. Analysis of calcium transients was performed using CytoSolver software (IonOptix LLC). Results: calcium transient parameters and frequency of Y4734C and WT were evaluated. Time to peak and Time constant were significantly longer in Y4734C (0,24±xx and 0,20±xx s, respectively; n=8; P˂0.004 and P˂0.048) than in WT (0,09±xx and 0,11±xx s, respectively; n=4). A nonsignificant change was observed in the amplitude of the calcium transient, Y4734C (0,22±xx; n=8) and WT (0,15±xx; n=4). Cell clusters with the variant Y4734C have higher frequency then WT (40±xx and 28±xx b/min, respectively). Conclusions: the preliminary data showed delayed release of calcium from the sarcoplasmic reticulum in cells with the variant Y4734C and prolonged reabsorption of calcium back into the sarcoplasmic reticulum.

Návaznosti

MUNI/A/1547/2023, interní kód MU

Název: Analýza (dys)funkce: od molekul k živému organismu

Investor: Masarykova univerzita, Analýza (dys)funkce: od molekul k živému organismu

NU22-02-00348, projekt VaV

Název: Funkční hodnocení genetických variant u případů klinicky „skutečné“ idiopatické fibrilace komor: in vitro a in silico modelování s cílem odhalit arytmogenní mechanismus

Investor: Ministerstvo zdravotnictví ČR, Funkční hodnocení genetických variant u případů klinicky „skutečné“ idiopatické fibrilace komor: in vitro a in silico modelování s cílem odhalit arytmogenní mechanismus, Podprogram 1 - standardní

Citovat

ŠVECOVÁ, Olga, Martin KRÁL, Štefan ZELENÁK, Jiří PACHERNÍK, Tomáš BÁRTA, Samuel LIETAVA, Iva SYNKOVÁ, Jana ZÍDKOVÁ, Tomáš NOVOTNÝ a Markéta BÉBAROVÁ. Analysis of calcium transients in cardiomyocytes derived from hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy controls. In 15th New Frontiers in Basic Cardiovascular Research: A France-New EU Members Symposium. 2024.

@proceedings{2417402,
   author = {Švecová, Olga and Král, Martin and Zelenák, Štefan and Pacherník, Jiří and Bárta, Tomáš and Lietava, Samuel and Synková, Iva and Zídková, Jana and Novotný, Tomáš and Bébarová, Markéta},
   booktitle = {15th New Frontiers in Basic Cardiovascular Research: A France-New EU Members Symposium},
   keywords = {calcium transient; hiPSC-derived cardiomyocytes},
   language = {eng},
   title = {Analysis of calcium transients in cardiomyocytes derived from hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy controls},
   year = {2024}
}

TY  - CONF
ID  - 2417402
AU  - Švecová, Olga - Král, Martin - Zelenák, Štefan - Pacherník, Jiří - Bárta, Tomáš - Lietava, Samuel - Synková, Iva - Zídková, Jana - Novotný, Tomáš - Bébarová, Markéta
PY  - 2024
TI  - Analysis of calcium transients in cardiomyocytes derived from hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy controls
KW  - calcium transient
KW  - hiPSC-derived cardiomyocytes
N2  - Introduction: the regulation of intracellular calcium levels is crucial for excitation-contraction coupling. The release of calcium into the intracellular space is controlled by the ryanodine receptor type 2 (RYR2) located on the sarcoplasmic reticulum. Dysfunction of RYR2 is involved in the pathogenesis of inherited and non-inherited diseases such as cardiac arrhythmias, ventricular fibrillation, ventricular tachycardia, etc. The variant p. Y4734C in RYR2 was found in a patient with idiopathic ventricular fibrillation without structural changes in the heart and signs of arrhythmia on clinical examination. Preliminary data show calcium transient of patient-specific cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM). Methods: calcium transients of hiPSC-CM (Y4734C) and hiPSC-CM unrelated healthy controls (WT) were measured using the Myocyte Calcium and Contractility System (IonOptix LLC). Cell clusters were loaded with Fura-2 (Molecular Probes, Invitrogen) at a final concentration of 1 mM. Cells were incubated for 15 min in Tyrode solution with 1 μmol/L Fura-2-am at 37 °C and then washed repeatedly with Tyrode solution followed by incubation for 10 min in Tyrode solution at 37 °C. Measurements were performed in Tyrode solution at 37 ± 0.5 °C. Cells were not stimulated. Analysis of calcium transients was performed using CytoSolver software (IonOptix LLC). Results: calcium transient parameters and frequency of Y4734C and WT were evaluated. Time to peak and Time constant were significantly longer in Y4734C (0,24±xx and 0,20±xx s, respectively; n=8; P˂0.004 and P˂0.048) than in WT (0,09±xx and 0,11±xx s, respectively; n=4). A nonsignificant change was observed in the amplitude of the calcium transient, Y4734C (0,22±xx; n=8) and WT (0,15±xx; n=4). Cell clusters with the variant Y4734C have higher frequency then WT (40±xx and 28±xx b/min, respectively). Conclusions: the preliminary data showed delayed release of calcium from the sarcoplasmic reticulum in cells with the variant Y4734C and prolonged reabsorption of calcium back into the sarcoplasmic reticulum.
ER  -

ŠVECOVÁ, Olga, Martin KRÁL, Štefan ZELENÁK, Jiří PACHERNÍK, Tomáš BÁRTA, Samuel LIETAVA, Iva SYNKOVÁ, Jana ZÍDKOVÁ, Tomáš NOVOTNÝ a Markéta BÉBAROVÁ. Analysis of calcium transients in cardiomyocytes derived from hiPSCs: the variant p. Y4734C in RYR2 vs. unrelated healthy controls. In \textit{15th New Frontiers in Basic Cardiovascular Research: A France-New EU Members Symposium}. 2024.

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