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

Autor(es):Mostrar menos -

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: Biochemical and Biophysical Research Communications; v. 533, n. 3, p. 376-382, DEC 10 2020.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 13/17368-0 - Genômica cardiovascular: mechanismos & novas terapias - CVGen mech2ther
• Beneficiário: José Eduardo Krieger
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 16/11321-0 - Estímulo mecânico como componente chave para a maturação e caracterização de cardiomiócitos humanos derivados de células-tronco pluripotentes induzidas.
• Beneficiário: Thayane Antoniolli Crestani
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 14/14086-7 - Transplante de cardiomiócitos derivados de iPSC pós-infarto do miocárdio em modelo suíno
• Beneficiário: Clara Steichen
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 18/16860-2 - Caracterização morfológica e funcional de cardiomiócitos humanos derivados de células-tronco pluripotentes induzidas após estímulo elétrico
• Beneficiário: Thayane Antoniolli Crestani
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado
• Processo FAPESP: 15/50216-5 - Caracterização da população ideal de células cardíacas derivadas de hiPSC para a regeneração cardíaca após infarto do miocárdio
• Beneficiário: José Eduardo Krieger
• Modalidade de apoio: Auxílio à Pesquisa - Parceria para Inovação Tecnológica - PITE