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

Author(s):Show less -

Crestani, Thayane ^[1] ; Steichen, Clara ^[1] ; Neri, Elida ^[1] ; Rodrigues, Mariliza ^[1] ; Fonseca-Alaniz, Miriam Helena ^[1] ; Ormrod, Beth ^{[2, 3]} ; Holt, Mark R. ^{[2, 3]} ; Pandey, Pragati ^[4] ; Harding, Sian ^[4] ; Ehler, Elisabeth ^{[2, 3]} ; Krieger, Jose E. ^[1]

Affiliation

^[1] Univ Sao Paulo, Heart Inst InCor, Med Sch, Sao Paulo, SP - Brazil

^[2] Kings Coll London, BHF Res Excellence Ctr, Sch Cardiovasc Med & Sci, London - England

^[3] Kings Coll London, Randall Ctr Cell & Mol Biophys, Sch Basic & Med Biosci, London - England

^[4] Imperial Coll London, Natl Heart & Lung Inst, London - England

• Document type: Journal article
• Source: Biochemical and Biophysical Research Communications; v. 533, n. 3, p. 376-382, DEC 10 2020.
• Web of Science Citations: 0

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 beta-catenin). We detected ES-induced CM maturation as indicated by increased Tnnil 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. (C) 2020 Elsevier Inc. All rights reserved. (AU)

• FAPESP's process: 13/17368-0 - Cardiovascular genomics: mechanisms & novel therapeutics - CVGen mech2ther
• Grantee: José Eduardo Krieger
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 16/11321-0 - Mechanical stimulus as a key component for maturation and characterization of human induced pluripotent stem cell-derived cardiomyocytes.
• Grantee: Thayane Antoniolli Crestani
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 14/14086-7 - Transplantation of pig iPSC-derived cardiac cells in a pig model of myocardial infarction
• Grantee: Clara Steichen
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 18/16860-2 - Morphological and functional characterization of human induced pluripotent stem cells derived cardiomyocytes after electrical stimulation
• Grantee: Thayane Antoniolli Crestani
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 15/50216-5 - Characterization of the optimal hiPSC-derived cardiac cell population for heart regeneration after myocardial infarction
• Grantee: José Eduardo Krieger
• Support Opportunities: Research Grants - Research Partnership for Technological Innovation - PITE