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Identification of circular RNAs regulating cardiomyocyte proliferation in neonatal pig hearts
Ling Tang, Verah Nyarige, Pengsheng Li, Junwen Wang, Wuqiang Zhu
Ling Tang, Verah Nyarige, Pengsheng Li, Junwen Wang, Wuqiang Zhu
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Research Article Cardiology

Identification of circular RNAs regulating cardiomyocyte proliferation in neonatal pig hearts

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Abstract

Little is known about the expression patterns and functions of circular RNAs (circRNAs) in the heart of large mammals. In this study, we examined the expression profiles of circRNAs, microRNAs (miRNAs), and messenger RNAs (mRNAs) in neonatal pig hearts. Pig heart samples collected on postnatal days 1 (P1), 3 (P3), 7 (P7), and 28 (P28) were sent for total RNA sequencing. Our data revealed a total of 7,000 circRNAs in the 24 pig hearts. Pathway enrichment analysis of hallmark gene sets demonstrated that differentially expressed circRNAs were engaged in different pathways. The most significant difference was observed between P1 and the other 3 groups (P3, P7, and P28) in pathways related to cell cycle and muscle development. Out of the 10 circRNAs that were validated through real-time quantitative PCR to verify their expression, 6 exhibited significant effects on cell cycle activity in human induced pluripotent stem cell–derived cardiomyocytes following small interfering RNA–mediated knockdown. circRNA-miRNA-mRNA networks were constructed to understand the potential mechanisms of circRNAs in the heart. In conclusion, our study provided a data set for exploring the roles of circRNAs in pig hearts. In addition, we identified several circRNAs that regulate cardiomyocyte cell cycle.

Authors

Ling Tang, Verah Nyarige, Pengsheng Li, Junwen Wang, Wuqiang Zhu

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Figure 5

circRNAs that modulate cardiomyocyte cell cycle.

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circRNAs that modulate cardiomyocyte cell cycle.
hiPSC-CMs at day 28 aft...
hiPSC-CMs at day 28 after initiation of cardiac differentiation were utilized. The efficiency of siRNA-based knockdown of circRNAs in hiPSC-CMs was determined by qRT-PCR. The relative expression levels of circRNAs were normalized to GAPDH reference gene. Cell number was evaluated by bioluminescence analysis. Data were presented as percentage. (A, C, E, G, I, and K) Evaluation of the expression of hsa-ABLIM1_0001, hsa-RNF13_0004, hsa-KIF1B_0001, hsa-MYOM1_0001, hsa-AC096949_0001, and hsa-PDLIM5_0001 in hiPSC-CMs after siRNA treatment. (B, D, F, H, J, and L) Bioluminescence assay and quantification of bioluminescent signal intensity in hiPSC-CMs with siRNA-based knockdown of the specific circRNAs. All data were presented as mean ± SEM. Statistical analysis was performed via 2-tailed Student’s t test. n = 3 technical replicates in each group for panels A, C, E, G, I, and K. n = 4 technical replicates in each group for panels B, D, F, H, J, and L. **P < 0.01, ***P < 0.001, and ****P < 0.0001.

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ISSN 2379-3708

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