Electrical stimulation applied during differentiation drives the hiPSC-CMs towards a mature cardiac conduction-like cells

Thayane Crestani; Clara Steichen; Elida Neri; Mariliza Rodrigues; Miriam Helena Fonseca-Alaniz; Beth Ormrod; Mark R. Holt; Pragati Pandey; Sian Harding; Elisabeth Ehler; Jose E. Krieger

doi:10.1016/j.bbrc.2020.09.021

Electrical stimulation applied during differentiation drives the hiPSC-CMs towards a mature cardiac conduction-like cells

Thayane Crestani, Clara Steichen, Elida Neri, Mariliza Rodrigues, Miriam Helena Fonseca-Alaniz, Beth Ormrod, Mark R. Holt, Pragati Pandey, Sian Harding, Elisabeth Ehler, Jose E. Krieger^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

21 Citations (Scopus)

Abstract

Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) resemble fetal cardiomyocytes and electrical stimulation (ES) has been explored to mature the differentiated cells. Here, we hypothesize that ES applied at the beginning of the differentiation process, triggers both differentiation of the hiPSC-CMs into a specialized conduction system (CS) phenotype and cell maturation. We applied ES for 15 days starting on day 0 of the differentiation process and found an increased expression of transcription factors and proteins associated with the development and function of CS including Irx3, Nkx2.5 and contactin 2, Hcn4 and Scn5a, respectively. We also found activation of intercalated disc proteins (Nrap and β-catenin). We detected ES-induced CM maturation as indicated by increased Tnni1 and Tnni3 expression. Confocal micrographs showed a shift towards expression of the gap junction protein connexin 40 in ES hiPSC-CM compared to the more dominant expression of connexin 43 in controls. Finally, analysis of functional parameters revealed that ES hiPSC-CMs exhibited faster action potential (AP) depolarization, longer intracellular Ca²⁺ transients, and slower AP duration at 90% of repolarization, resembling fast conducting fibers. Altogether, we provided evidence that ES during the differentiation of hiPSC to cardiomyocytes lead to development of cardiac conduction-like cells with more mature cytoarchitecture. Thus, hiPSC-CMs exposed to ES during differentiation can be instrumental to develop CS cells for cardiac disease modelling, screening individual drugs on a precison medicine type platform and support the development of novel therapeutics for arrhythmias.

Original language	English
Pages (from-to)	376-382
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	533
Issue number	3
DOIs	https://doi.org/10.1016/j.bbrc.2020.09.021
Publication status	Published - 10 Dec 2020

Keywords

Differentiation
Electrical stimulation
Human induced pluripotent stem cells derived cardiomyocytes
Maturation
Specialized cardiac conduction cells

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.bbrc.2020.09.021

Cite this

Crestani, T., Steichen, C., Neri, E., Rodrigues, M., Fonseca-Alaniz, M. H., Ormrod, B., Holt, M. R., Pandey, P., Harding, S., Ehler, E., & Krieger, J. E. (2020). Electrical stimulation applied during differentiation drives the hiPSC-CMs towards a mature cardiac conduction-like cells. Biochemical and Biophysical Research Communications, 533(3), 376-382. https://doi.org/10.1016/j.bbrc.2020.09.021

@article{e426a9bc6dc1455fbf297dd3910152cf,

title = "Electrical stimulation applied during differentiation drives the hiPSC-CMs towards a mature cardiac conduction-like cells",

abstract = "Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) resemble fetal cardiomyocytes and electrical stimulation (ES) has been explored to mature the differentiated cells. Here, we hypothesize that ES applied at the beginning of the differentiation process, triggers both differentiation of the hiPSC-CMs into a specialized conduction system (CS) phenotype and cell maturation. We applied ES for 15 days starting on day 0 of the differentiation process and found an increased expression of transcription factors and proteins associated with the development and function of CS including Irx3, Nkx2.5 and contactin 2, Hcn4 and Scn5a, respectively. We also found activation of intercalated disc proteins (Nrap and β-catenin). We detected ES-induced CM maturation as indicated by increased Tnni1 and Tnni3 expression. Confocal micrographs showed a shift towards expression of the gap junction protein connexin 40 in ES hiPSC-CM compared to the more dominant expression of connexin 43 in controls. Finally, analysis of functional parameters revealed that ES hiPSC-CMs exhibited faster action potential (AP) depolarization, longer intracellular Ca2+ transients, and slower AP duration at 90% of repolarization, resembling fast conducting fibers. Altogether, we provided evidence that ES during the differentiation of hiPSC to cardiomyocytes lead to development of cardiac conduction-like cells with more mature cytoarchitecture. Thus, hiPSC-CMs exposed to ES during differentiation can be instrumental to develop CS cells for cardiac disease modelling, screening individual drugs on a precison medicine type platform and support the development of novel therapeutics for arrhythmias.",

keywords = "Differentiation, Electrical stimulation, Human induced pluripotent stem cells derived cardiomyocytes, Maturation, Specialized cardiac conduction cells",

author = "Thayane Crestani and Clara Steichen and Elida Neri and Mariliza Rodrigues and Fonseca-Alaniz, {Miriam Helena} and Beth Ormrod and Holt, {Mark R.} and Pragati Pandey and Sian Harding and Elisabeth Ehler and Krieger, {Jose E.}",

year = "2020",

month = dec,

day = "10",

doi = "10.1016/j.bbrc.2020.09.021",

language = "English",

volume = "533",

pages = "376--382",

journal = "Biochemical and Biophysical Research Communications",

issn = "0006-291X",

publisher = "ACADEMIC PRESS INC",

number = "3",

}

Crestani, T, Steichen, C, Neri, E, Rodrigues, M, Fonseca-Alaniz, MH, Ormrod, B , Holt, MR, Pandey, P, Harding, S, Ehler, E & Krieger, JE 2020, 'Electrical stimulation applied during differentiation drives the hiPSC-CMs towards a mature cardiac conduction-like cells', Biochemical and Biophysical Research Communications, vol. 533, no. 3, pp. 376-382. https://doi.org/10.1016/j.bbrc.2020.09.021

TY - JOUR

T1 - Electrical stimulation applied during differentiation drives the hiPSC-CMs towards a mature cardiac conduction-like cells

AU - Crestani, Thayane

AU - Steichen, Clara

AU - Neri, Elida

AU - Rodrigues, Mariliza

AU - Fonseca-Alaniz, Miriam Helena

AU - Ormrod, Beth

AU - Holt, Mark R.

AU - Pandey, Pragati

AU - Harding, Sian

AU - Ehler, Elisabeth

AU - Krieger, Jose E.

PY - 2020/12/10

Y1 - 2020/12/10

N2 - Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) resemble fetal cardiomyocytes and electrical stimulation (ES) has been explored to mature the differentiated cells. Here, we hypothesize that ES applied at the beginning of the differentiation process, triggers both differentiation of the hiPSC-CMs into a specialized conduction system (CS) phenotype and cell maturation. We applied ES for 15 days starting on day 0 of the differentiation process and found an increased expression of transcription factors and proteins associated with the development and function of CS including Irx3, Nkx2.5 and contactin 2, Hcn4 and Scn5a, respectively. We also found activation of intercalated disc proteins (Nrap and β-catenin). We detected ES-induced CM maturation as indicated by increased Tnni1 and Tnni3 expression. Confocal micrographs showed a shift towards expression of the gap junction protein connexin 40 in ES hiPSC-CM compared to the more dominant expression of connexin 43 in controls. Finally, analysis of functional parameters revealed that ES hiPSC-CMs exhibited faster action potential (AP) depolarization, longer intracellular Ca2+ transients, and slower AP duration at 90% of repolarization, resembling fast conducting fibers. Altogether, we provided evidence that ES during the differentiation of hiPSC to cardiomyocytes lead to development of cardiac conduction-like cells with more mature cytoarchitecture. Thus, hiPSC-CMs exposed to ES during differentiation can be instrumental to develop CS cells for cardiac disease modelling, screening individual drugs on a precison medicine type platform and support the development of novel therapeutics for arrhythmias.

AB - Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) resemble fetal cardiomyocytes and electrical stimulation (ES) has been explored to mature the differentiated cells. Here, we hypothesize that ES applied at the beginning of the differentiation process, triggers both differentiation of the hiPSC-CMs into a specialized conduction system (CS) phenotype and cell maturation. We applied ES for 15 days starting on day 0 of the differentiation process and found an increased expression of transcription factors and proteins associated with the development and function of CS including Irx3, Nkx2.5 and contactin 2, Hcn4 and Scn5a, respectively. We also found activation of intercalated disc proteins (Nrap and β-catenin). We detected ES-induced CM maturation as indicated by increased Tnni1 and Tnni3 expression. Confocal micrographs showed a shift towards expression of the gap junction protein connexin 40 in ES hiPSC-CM compared to the more dominant expression of connexin 43 in controls. Finally, analysis of functional parameters revealed that ES hiPSC-CMs exhibited faster action potential (AP) depolarization, longer intracellular Ca2+ transients, and slower AP duration at 90% of repolarization, resembling fast conducting fibers. Altogether, we provided evidence that ES during the differentiation of hiPSC to cardiomyocytes lead to development of cardiac conduction-like cells with more mature cytoarchitecture. Thus, hiPSC-CMs exposed to ES during differentiation can be instrumental to develop CS cells for cardiac disease modelling, screening individual drugs on a precison medicine type platform and support the development of novel therapeutics for arrhythmias.

KW - Differentiation

KW - Electrical stimulation

KW - Human induced pluripotent stem cells derived cardiomyocytes

KW - Maturation

KW - Specialized cardiac conduction cells

UR - http://www.scopus.com/inward/record.url?scp=85091256277&partnerID=8YFLogxK

U2 - 10.1016/j.bbrc.2020.09.021

DO - 10.1016/j.bbrc.2020.09.021

M3 - Article

AN - SCOPUS:85091256277

SN - 0006-291X

VL - 533

SP - 376

EP - 382

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 3

ER -

Electrical stimulation applied during differentiation drives the hiPSC-CMs towards a mature cardiac conduction-like cells

Abstract

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Cite this