Functional screening identifies miRNAs inducing cardiac regeneration

Abstract

In mammals, enlargement of the heart during embryonic development is primarily dependent on the increase in cardiomyocyte numbers. Shortly after birth, however, cardiomyocytes stop proliferating and further growth of the myocardium occurs through hypertrophic enlargement of the existing myocytes. As a consequence of the minimal renewal of cardiomyocytes during adult life, repair of cardiac damage through myocardial regeneration is very limited. Here we show that the exogenous administration of selected microRNAs (miRNAs) markedly stimulates cardiomyocyte proliferation and promotes cardiac repair. We performed a high-content microscopy, high-throughput functional screening for human miRNAs that promoted neonatal cardiomyocyte proliferation using a whole-genome miRNA library. Forty miRNAs strongly increased both DNA synthesis and cytokinesis in neonatal mouse and rat cardiomyocytes. Two of these miRNAs (hsa-miR-590 and hsa-miR-199a) were further selected for testing and were shown to promote cell cycle re-entry of adult cardiomyocytes ex vivo and to promote cardiomyocyte proliferation in both neonatal and adult animals. After myocardial infarction in mice, these miRNAs stimulated marked cardiac regeneration and almost complete recovery of cardiac functional parameters. The miRNAs identified hold great promise for the treatment of cardiac pathologies consequent to cardiomyocyte loss.

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Figure 1: High-content screening identifies miRNAs regulating CM proliferation.
Figure 2: miRNAs increase cytokinesis and proliferation of post-natal CMs.
Figure 3: Genes downregulated by miR-590-3p and miR-199a-3p increase CM proliferation.
Figure 4: miR-590 and miR-199a induce CM proliferation in vivo.
Figure 5: miR-590-3p and miR-199a-3p induce marked cardiac regeneration after myocardial infarction.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

The cardiomyocyte miRNA expression microarray and transcriptomic data are deposited at GEO, under accession numbers GSE41537 and GSE41538, respectively.

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Acknowledgements

The authors are grateful to M. Dapas and M. Zotti for technical support in AAV production, to M. Sturnega for help in animal experimentation and to S. Kerbavcic for editorial assistance. A.E. is recipient of a FEBS Long Term Fellowship. This work was supported by Advanced Grant 250124 from the European Research Council (ERC) to M.G. and from Project CTC from the Fondazione CRTrieste, Trieste, Italy.

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A.E., M.M. and M.G. designed the study. A.E., M.M., S.Z. and M.D.F. performed the experiments and analysed the data. G.S. provided clinical consultancy for the animal study. L.Z. was responsible for AAV production. A.E., M.M. and M.G. wrote the manuscript.

Corresponding author

Correspondence to Mauro Giacca.

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Competing interests

A.E., M.M., S.Z. and M.G. are listed as co-inventors on a pending patent held by ICGEB that relates to the clinical development of the miRNAs promoting cardiac proliferation described in this paper.

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Eulalio, A., Mano, M., Ferro, M. et al. Functional screening identifies miRNAs inducing cardiac regeneration. Nature 492, 376–381 (2012). https://doi.org/10.1038/nature11739

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