| Full text | |
| Author(s): Show less - |
Munoz, Juan J. A. M.
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Dariolli, Rafael
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da Silva, Caio Mateus
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Neri, Elida A.
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Valadao, Iuri C.
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Turaca, Lauro Thiago
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Lima, Vanessa M.
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Peres de Carvalho, Mariana Lombardi
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Velho, Mariliza R.
;
Sobie, Eric A.
;
Krieger, Jose E.
Total Authors: 11
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| Document type: | Journal article |
| Source: | STEM CELL RESEARCH & THERAPY; v. 13, n. 1, p. 27-pg., 2022-09-02. |
| Abstract | |
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) | |
| FAPESP's process: | 16/07541-5 - Action potential and Calcium transient phenotyping of cardiomyocytes derived from pluripotent stem cells: development of an electrophysiological platform CM-PSCs compatible to study disease and drug modeling and cardiac regeneration approaches |
| Grantee: | Rafael Dariolli |
| Support Opportunities: | Scholarships abroad - Research Internship - Post-doctor |
| FAPESP's process: | 14/50889-7 - National Institute of Science and Technology Medicine Assisted by Scientific Computing (INCT-MACC) |
| Grantee: | José Eduardo Krieger |
| Support Opportunities: | Research Projects - Thematic Grants |
| 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: | 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 |
| FAPESP's process: | 14/24577-8 - Nanostructures and iPSC derived cardiomyocytes approaches for cardiac protection and repair post-MI |
| Grantee: | Rafael Dariolli |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |