Electrophysiological and calcium-handling development during long-term culture of human-induced pluripotent stem cell-derived cardiomyocytes

Seibertz, Fitzwilliam; Sutanto, Henry; Dülk, Rebekka; Pronto, Julius Ryan; Springer, Robin; Rapedius, Markus; Liutkute, Aiste; Ritter, Melanie; Jung, Philipp; Stelzer, Lea; Hüsgen, Luisa M; Klopp, Marie; Rubio, Tony; Fakuade, Funsho E; Mason, Fleur E.; Hartmann, Nico; Pabel, Steffen; Streckfuß‐Bömeke, Katrin; Cyganek, Lukas; Sossalla, Samuel; Heijman, Jordi; Voigt, Niels

doi:10.1007/s00395-022-00973-0

Cited by 16 publications

(9 citation statements)

References 81 publications

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“…In the in vitro experiments of the present study, NRCFs isolated from rats and MCFs isolated from mice were used to explore the effect of NPRC deficiency on CFs. To further verify the experimental results obtained from animal cells, CFs and CMs differentiated from hiPSCs were also used, which have been increasingly applied for cardiac disease modeling and drug screening ( 51 , 52 ). Here, NPRC deficiency in hiPSC-CFs activated cAMP/PKA and cGMP/PKG signaling, regulated TGIF1 and TGF-β1/Smad signaling, and decreased collagen synthesis under HG stimulation.…”

Section: Discussionmentioning

confidence: 99%

NPRC deletion attenuates cardiac fibrosis in diabetic mice by activating PKA/PKG and inhibiting TGF-β1/Smad pathways

Meng

Wang

et al. 2023

Sci. Adv.

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Cardiac fibrosis plays a key role in the progression of diabetic cardiomyopathy (DCM). Previous studies demonstrated the cardioprotective effects of natriuretic peptides. However, the effects of natriuretic peptide receptor C (NPRC) on cardiac fibrosis in DCM remains unknown. Here, we observed that myocardial NPRC expression was increased in mice and patients with DCM. NPRC −/− diabetic mice showed alleviated cardiac fibrosis, as well as improved cardiac function and remodeling. NPRC knockdown in both cardiac fibroblasts and cardiomyocytes decreased collagen synthesis and proliferation of cardiac fibroblasts. RNA sequencing identified that NPRC deletion up-regulated the expression of TGF-β–induced factor homeobox 1 (TGIF1), which inhibited the phosphorylation of Smad2/3. Furthermore, TGIF1 up-regulation was mediated by the activation of cAMP/PKA and cGMP/PKG signaling induced by NPRC deletion. These findings suggest that NPRC deletion attenuated cardiac fibrosis and improved cardiac remodeling and function in diabetic mice, providing a promising approach to the treatment of diabetic cardiac fibrosis.

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Section: Discussionmentioning

confidence: 99%

NPRC deletion attenuates cardiac fibrosis in diabetic mice by activating PKA/PKG and inhibiting TGF-β1/Smad pathways

Meng

Wang

et al. 2023

Sci. Adv.

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“…HiPSCs were differentiated in atrial- and ventricular-like hiPSC-derived cardiomyocytes (hiPSC-CMs) following previously published protocols [ 43 ]. Automated patch clamp (APC) experiments were performed using the SyncroPatch 384 APC System (Nanion Technologies, Munich, Germany) as previously described [ 44 ]. Electrical activity of hiPSC-CM monolayers was assessed using the Maestro Pro multi-electrode array (MEA) system (Axion Biosystems, Atlanta, USA) as described in the Supplemental Material online.…”

Section: Methodsmentioning

confidence: 99%

Acute antiarrhythmic effects of SGLT2 inhibitors–dapagliflozin lowers the excitability of atrial cardiomyocytes

Paasche,

Wiedmann,

Kraft

et al. 2024

Basic Res Cardiol

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In recent years, SGLT2 inhibitors have become an integral part of heart failure therapy, and several mechanisms contributing to cardiorenal protection have been identified. In this study, we place special emphasis on the atria and investigate acute electrophysiological effects of dapagliflozin to assess the antiarrhythmic potential of SGLT2 inhibitors. Direct electrophysiological effects of dapagliflozin were investigated in patch clamp experiments on isolated atrial cardiomyocytes. Acute treatment with elevated-dose dapagliflozin caused a significant reduction of the action potential inducibility, the amplitude and maximum upstroke velocity. The inhibitory effects were reproduced in human induced pluripotent stem cell-derived cardiomyocytes, and were more pronounced in atrial compared to ventricular cells. Hypothesizing that dapagliflozin directly affects the depolarization phase of atrial action potentials, we examined fast inward sodium currents in human atrial cardiomyocytes and found a significant decrease of peak sodium current densities by dapagliflozin, accompanied by a moderate inhibition of the transient outward potassium current. Translating these findings into a porcine large animal model, acute elevated-dose dapagliflozin treatment caused an atrial-dominant reduction of myocardial conduction velocity in vivo. This could be utilized for both, acute cardioversion of paroxysmal atrial fibrillation episodes and rhythm control of persistent atrial fibrillation. In this study, we show that dapagliflozin alters the excitability of atrial cardiomyocytes by direct inhibition of peak sodium currents. In vivo, dapagliflozin exerts antiarrhythmic effects, revealing a potential new additional role of SGLT2 inhibitors in the treatment of atrial arrhythmias.

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“…Stable human embryonic kidney (HEK)-293- or Chinese hamster ovary (CHO) cell lines overexpressing hERG1a display large current densities that can be reliably studied. In contrast, hERG1a expression levels in native cardiomyocytes and hiPSC-derived cardiomyocytes are very low ( Goversen et al, 2019 ; Seibertz et al, 2023b ). Normalized to membrane capacitance, a maximal I Kr density is in the order of 1 pA/pF, which translates to absolute values of ∼150 pA and ∼30 pA in adult cardiomyocytes and hiPSC-CM, respectively.…”

Section: Introductionmentioning

confidence: 99%

Recording ten-fold larger IKr conductances with automated patch clamping using equimolar Cs+ solutions

Bloothooft,

Verbruggen,

Seibertz

et al. 2024

Front. Physiol.

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Background: The rapid delayed rectifier potassium current (IKr) is important for cardiac repolarization and is most often involved in drug-induced arrhythmias. However, accurately measuring this current can be challenging in human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes because of its small current density. Interestingly, the ion channel conducting IKr, hERG channel, is not only permeable to K+ ions but also to Cs+ ions when present in equimolar concentrations inside and outside of the cell.Methods: In this study, IhERG was measured from Chinese hamster ovary (CHO)-hERG cells and hiPSC-CM using either Cs+ or K+ as the charge carrier. Equimolar Cs+ has been used in the literature in manual patch-clamp experiments, and here, we apply this approach using automated patch-clamp systems. Four different (pre)clinical drugs were tested to compare their effects on Cs+- and K+-based currents.Results: Using equimolar Cs+ solutions gave rise to approximately ten-fold larger hERG conductances. Comparison of Cs+- and K+-mediated currents upon application of dofetilide, desipramine, moxifloxacin, or LUF7244 revealed many similarities in inhibition or activation properties of the drugs studied. Using equimolar Cs+ solutions gave rise to approximately ten-fold larger hERG conductances. In hiPSC-CM, the Cs+-based conductance is larger compared to the known K+-based conductance, and the Cs+ hERG conductance can be inhibited similarly to the K+-based conductance.Conclusion: Using equimolar Cs+ instead of K+ for IhERG measurements in an automated patch-clamp system gives rise to a new method by which, for example, quick scans can be performed on effects of drugs on hERG currents. This application is specifically relevant when such experiments are performed using cells which express small IKr current densities in combination with small membrane capacitances.

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Electrophysiological and calcium-handling development during long-term culture of human-induced pluripotent stem cell-derived cardiomyocytes

Cited by 16 publications

References 81 publications

NPRC deletion attenuates cardiac fibrosis in diabetic mice by activating PKA/PKG and inhibiting TGF-β1/Smad pathways

NPRC deletion attenuates cardiac fibrosis in diabetic mice by activating PKA/PKG and inhibiting TGF-β1/Smad pathways

Acute antiarrhythmic effects of SGLT2 inhibitors–dapagliflozin lowers the excitability of atrial cardiomyocytes

Recording ten-fold larger IKr conductances with automated patch clamping using equimolar Cs+ solutions

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