Time-regulated transcripts with the potential to modulate human pluripotent stem cell-derived cardiomyocyte differentiation

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Munoz, Juan J. A. M. ; Dariolli, Rafael ; da Silva, Caio Mateus ; Neri, Elida A. ; Valadao, Iuri C. ; Turaca, Lauro Thiago ; Lima, Vanessa M. ; Peres de Carvalho, Mariana Lombardi ; Velho, Mariliza R. ; Sobie, Eric A. ; Krieger, Jose E.

• Tipo de documento: Artigo Científico
• Fonte: STEM CELL RESEARCH & THERAPY; v. 13, n. 1, p. 27-pg., 2022-09-02.

Resumo

Background Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) are a promising disease model, even though hiPSC-CMs cultured for extended periods display an undifferentiated transcriptional landscape. MiRNA-target gene interactions contribute to fine-tuning the genetic program governing cardiac maturation and may uncover critical pathways to be targeted. Methods We analyzed a hiPSC-CM public dataset to identify time-regulated miRNA-target gene interactions based on three logical steps of filtering. We validated this process in silico using 14 human and mouse public datasets, and further confirmed the findings by sampling seven time points over a 30-day protocol with a hiPSC-CM clone developed in our laboratory. We then added miRNA mimics from the top eight miRNAs candidates in three cell clones in two different moments of cardiac specification and maturation to assess their impact on differentiation characteristics including proliferation, sarcomere structure, contractility, and calcium handling. Results We uncovered 324 interactions among 29 differentially expressed genes and 51 miRNAs from 20,543 transcripts through 120 days of hiPSC-CM differentiation and selected 16 genes and 25 miRNAs based on the inverse pattern of expression (Pearson R-values < - 0.5) and consistency in different datasets. We validated 16 inverse interactions among eight genes and 12 miRNAs (Person R-values < - 0.5) during hiPSC-CMs differentiation and used miRNAs mimics to verify proliferation, structural and functional features related to maturation. We also demonstrated that miR-124 affects Ca2+ handling altering features associated with hiPSC-CMs maturation. Conclusion We uncovered time-regulated transcripts influencing pathways affecting cardiac differentiation/maturation axis and showed that the top-scoring miRNAs indeed affect primarily structural features highlighting their role in the hiPSC-CM maturation. (AU)

• Processo FAPESP: 16/07541-5 - Fenotipagem de cardiomiócitos derivados de células-tronco pluripotentes através da medida de potencial de ação e transientes de cálcio: desenvolvimento de uma plataforma eletrofisiológica CM-PSCs compatível
• Beneficiário: Rafael Dariolli
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 14/50889-7 - INCT 2014: em Medicina Assistida por Computação Científica (INCT-MACC)
• Beneficiário: José Eduardo Krieger
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• 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: 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
• Processo FAPESP: 14/24577-8 - Uso de microscaffolds (lockyballs) e cardiomiocitos derivados de iPSC para proteção e reparação cardíaca pós-infarto do miocárdio
• Beneficiário: Rafael Dariolli
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado