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An endothelial apelin-FGF link mediated by miR-424 and miR-503 is disrupted in pulmonary arterial hypertension

Abstract

Pulmonary arterial hypertension (PAH) is characterized by vascular remodeling associated with obliteration of pulmonary arterioles and formation of plexiform lesions composed of hyperproliferative endothelial and vascular smooth-muscle cells. Here we describe a microRNA (miRNA)-dependent association between apelin (APLN) and fibroblast growth factor 2 (FGF2) signaling in pulmonary artery endothelial cells (PAECs). APLN deficiency in these cells led to increased expression of FGF2 and its receptor FGFR1 as a consequence of decreased expression of miR-424 and miR-503, which directly target FGF2 and FGFR1. miR-424 and miR-503 were downregulated in PAH, exerted antiproliferative effects in PAECs and inhibited the capacity of PAEC-conditioned medium to induce the proliferation of pulmonary artery smooth-muscle cells. Reconstitution of miR-424 and miR-503 in vivo ameliorated pulmonary hypertension in experimental models. These studies identify an APLN-dependent miRNA-FGF signaling axis needed for the maintenance of pulmonary vascular homeostasis.

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Figure 1: APLN downregulates FGF2 expression in PAECs in an miRNA-dependent manner.
Figure 2: The APLN-FGF2 axis is mediated by APLN-regulated expression of miR-424 and miR-503.
Figure 3: miR-424 and miR-503 regulate FGF2 and FGFR1 expression and signaling.
Figure 4: Downregulation of miR-424 and miR-503 in PAH is associated with increased FGF2 and FGFR1 expression.
Figure 5: Endothelial miR-424 and miR-503 regulate PAEC proliferation and migration and induce paracrine inhibition of PASMC proliferation.
Figure 6: miR-424 and miR-503 overexpression ameliorate experimental pulmonary hypertension models in rats.

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Acknowledgements

We thank M. Simons and P. Yu for critical reading of the manuscript, R. Homer for pathology slide review, T. Quertermous (Stanford University) for the Apln-null mice, P. Lee and Y. Zhang for guidance with the lentiviral work and the Yale Center for Genome Analysis for miRNA array analyses. This study was supported by grants from the US National Institutes of Health (HL095654, HL113005 and HL101284 to H.J.C., HL069170 to S.C.E. and HL093362 to D.M.G.), the Howard Hughes Medical Institute (Physician Scientist Early Career Award to H.J.C.), an American Heart Association Grant-in-Aid (12GRNT9410029 to H.J.C.) and the Pfizer ASPIRE Young Investigator Research Award (H.J.C.).

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J.K. and H.J.C. designed the research. J.K., Y. Kang, Y. Kojima, J.K.L., X.H., D.L.M., H.P. and H.J.C. performed the experiments. M.A.A., S.A.C. and S.C.E. collected and prepared the subject samples. D.M.G. and S.C.E. assisted with data analysis and review of the manuscript. J.K. and H.J.C. prepared the figures. J.K. and H.J.C. wrote the manuscript.

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Correspondence to Hyung J Chun.

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Kim, J., Kang, Y., Kojima, Y. et al. An endothelial apelin-FGF link mediated by miR-424 and miR-503 is disrupted in pulmonary arterial hypertension. Nat Med 19, 74–82 (2013). https://doi.org/10.1038/nm.3040

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