Spontaneous electrical activity of patient-specific cardiomyocytes with a variant in the RYR2 gene recorded by multielectrode array technique

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14110%2F24%3A00137555" target="_blank" >RIV/00216224:14110/24:00137555 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Spontaneous electrical activity of patient-specific cardiomyocytes with a variant in the RYR2 gene recorded by multielectrode array technique

Popis výsledku v původním jazyce

Background: Y4734C variant in the ryanodine receptor type 2 (RYR2) was found in a patient with idiopathic ventricular fibrillation and his sister who was diagnosed with catecholaminergic polymorphic ventricular tachycardia (CPVT). The variant hasn´t been functionally tested before. To reveal the proarrhythmic potential of the variant, cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM) of the proband (IF), his sister (CPVT), and his healthy nephew as control (WT) were prepared. Here we bring data showing the spontaneous electrical activity of the hiPSC-CM and its changes under β-adrenergic stimulation. Methods: The spontaneous electrical activity of hiPSC-CM in control conditions (CC) and under the effect of 0.5µM isoprenaline (ISO) stimulating β-adrenergic receptors was recorded by the multielectrode array technique at 37C. The cycle length (CL) and field potential duration (FPD) were evaluated; Bazett´s formula was used to correct FPD (FPDc). The short-term variability of CL (STVCL) was calculated using Origin2024 software. The data are presented as the median±interquartile range of n samples (nWT=11; nIF=19; nCPVT=18), and non-parametric tests (Wilcoxon and Kruskal-Wallis) were used due to non-normal data distribution; P˂0.05 was considered statistically significant. Results: CL was significantly shorter in WT (0.71±0.29s) than in CPVT (1.40±0.81s); IF did not differ from both (1.10±1.23s). There was no difference between WT, IF, and CPVT in FPD (152.1±106.3, 148.5±75.9, and 128.0±46.0ms, respectively), however, FPDc was significantly longer in WT (179.1±139.1ms) than in CPVT (94.4±41.7ms). After adding ISO, CL and FPD were significantly shortened in all groups (WT CLISO=0.62±0.20s, FPDISO=135.2±58.3ms; IF CLISO=0.67±0.84s, FPDISO=118.7±71.8ms; CPVT CLISO=1.12±0.80s, FPDISO=104.4±25.7ms); no significant changes were observed in FPDc. The CPVT showed significantly larger shortening in CL than WT and IF (by 38.7±21.6, 13.8±23.3, and 25.7±28.5%, respectively) whereas no differences were apparent in FPD shortening. IF and CPVT showed significantly larger STVCL than WT in CC (18.3±45.1, 34.0±42.6, and 3.9±13.2ms, respectively). ISO significantly decreased STVCL in both IF and CPVT (9.0±27.8 and 8.5±7.8ms, respectively) but had no significant effect on WT (8.1±42.7ms). Conclusions: CPVT showed a slower beating rate than WT and higher reactivity to ISO than WT and IF. ISO increased beating frequency and shortened FPD in every group. IF and CPVT had higher beat-to-beat variability of CL than WT in CC, but no difference among groups was apparent under ISO.

Název v anglickém jazyce

Spontaneous electrical activity of patient-specific cardiomyocytes with a variant in the RYR2 gene recorded by multielectrode array technique

Popis výsledku anglicky

Background: Y4734C variant in the ryanodine receptor type 2 (RYR2) was found in a patient with idiopathic ventricular fibrillation and his sister who was diagnosed with catecholaminergic polymorphic ventricular tachycardia (CPVT). The variant hasn´t been functionally tested before. To reveal the proarrhythmic potential of the variant, cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM) of the proband (IF), his sister (CPVT), and his healthy nephew as control (WT) were prepared. Here we bring data showing the spontaneous electrical activity of the hiPSC-CM and its changes under β-adrenergic stimulation. Methods: The spontaneous electrical activity of hiPSC-CM in control conditions (CC) and under the effect of 0.5µM isoprenaline (ISO) stimulating β-adrenergic receptors was recorded by the multielectrode array technique at 37C. The cycle length (CL) and field potential duration (FPD) were evaluated; Bazett´s formula was used to correct FPD (FPDc). The short-term variability of CL (STVCL) was calculated using Origin2024 software. The data are presented as the median±interquartile range of n samples (nWT=11; nIF=19; nCPVT=18), and non-parametric tests (Wilcoxon and Kruskal-Wallis) were used due to non-normal data distribution; P˂0.05 was considered statistically significant. Results: CL was significantly shorter in WT (0.71±0.29s) than in CPVT (1.40±0.81s); IF did not differ from both (1.10±1.23s). There was no difference between WT, IF, and CPVT in FPD (152.1±106.3, 148.5±75.9, and 128.0±46.0ms, respectively), however, FPDc was significantly longer in WT (179.1±139.1ms) than in CPVT (94.4±41.7ms). After adding ISO, CL and FPD were significantly shortened in all groups (WT CLISO=0.62±0.20s, FPDISO=135.2±58.3ms; IF CLISO=0.67±0.84s, FPDISO=118.7±71.8ms; CPVT CLISO=1.12±0.80s, FPDISO=104.4±25.7ms); no significant changes were observed in FPDc. The CPVT showed significantly larger shortening in CL than WT and IF (by 38.7±21.6, 13.8±23.3, and 25.7±28.5%, respectively) whereas no differences were apparent in FPD shortening. IF and CPVT showed significantly larger STVCL than WT in CC (18.3±45.1, 34.0±42.6, and 3.9±13.2ms, respectively). ISO significantly decreased STVCL in both IF and CPVT (9.0±27.8 and 8.5±7.8ms, respectively) but had no significant effect on WT (8.1±42.7ms). Conclusions: CPVT showed a slower beating rate than WT and higher reactivity to ISO than WT and IF. ISO increased beating frequency and shortened FPD in every group. IF and CPVT had higher beat-to-beat variability of CL than WT in CC, but no difference among groups was apparent under ISO.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

30201 - Cardiac and Cardiovascular systems

Projekt

<a href="/cs/project/NU22-02-00348" target="_blank" >NU22-02-00348: Funkční hodnocení genetických variant u případů klinicky „skutečné“ idiopatické fibrilace komor: in vitro a in silico modelování s cílem odhalit arytmogenní mechanismus</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů