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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
References
Tuder, R.M., Marecki, J.C., Richter, A., Fijalkowska, I. & Flores, S. Pathology of pulmonary hypertension. Clin. Chest Med. 28, 23–42, vii (2007).
Humbert, M. et al. Cellular and molecular pathobiology of pulmonary arterial hypertension. J. Am. Coll. Cardiol. 43, 13S–24S (2004).
Hassoun, P.M. et al. Inflammation, growth factors, and pulmonary vascular remodeling. J. Am. Coll. Cardiol. 54, S10–S19 (2009).
Schermuly, R.T., Ghofrani, H.A., Wilkins, M.R. & Grimminger, F. Mechanisms of disease: pulmonary arterial hypertension. Nat. Rev. Cardiol. 8, 443–455 (2011).
Izikki, M. et al. Endothelial-derived FGF2 contributes to the progression of pulmonary hypertension in humans and rodents. J. Clin. Invest. 119, 512–523 (2009).
Eddahibi, S. et al. Cross talk between endothelial and smooth muscle cells in pulmonary hypertension: critical role for serotonin-induced smooth muscle hyperplasia. Circulation 113, 1857–1864 (2006).
Dewachter, L. et al. Angiopoietin/Tie2 pathway influences smooth muscle hyperplasia in idiopathic pulmonary hypertension. Am. J. Respir. Crit. Care Med. 174, 1025–1033 (2006).
Chandra, S.M. et al. Disruption of the apelin-APJ system worsens hypoxia-induced pulmonary hypertension. Arterioscler. Thromb. Vasc. Biol. 31, 814–820 (2011).
Alastalo, T.P. et al. Disruption of PPARγ/β-catenin-mediated regulation of apelin impairs BMP-induced mouse and human pulmonary arterial EC survival. J. Clin. Invest. 121, 3735–3746 (2011).
Sheikh, A.Y. et al. In vivo genetic profiling and cellular localization of apelin reveals a hypoxia-sensitive, endothelial-centered pathway activated in ischemic heart failure. Am. J. Physiol. Heart Circ. Physiol. 294, H88–H98 (2008).
Hosoya, M. et al. Molecular and functional characteristics of APJ. Tissue distribution of mRNA and interaction with the endogenous ligand apelin. J. Biol. Chem. 275, 21061–21067 (2000).
Regard, J.B., Sato, I.T. & Coughlin, S.R. Anatomical profiling of G protein–coupled receptor expression. Cell 135, 561–571 (2008).
Goetze, J.P. et al. Apelin: a new plasma marker of cardiopulmonary disease. Regul. Pept. 133, 134–138 (2006).
Tuder, R.M., Groves, B., Badesch, D.B. & Voelkel, N.F. Exuberant endothelial cell growth and elements of inflammation are present in plexiform lesions of pulmonary hypertension. Am. J. Pathol. 144, 275–285 (1994).
Masri, F.A. et al. Hyperproliferative apoptosis-resistant endothelial cells in idiopathic pulmonary arterial hypertension. Am. J. Physiol. Lung Cell. Mol. Physiol. 293, L548–L554 (2007).
Benisty, J.I. et al. Elevated basic fibroblast growth factor levels in patients with pulmonary arterial hypertension. Chest 126, 1255–1261 (2004).
Falcão-Pires, I. et al. Apelin decreases myocardial injury and improves right ventricular function in monocrotaline-induced pulmonary hypertension. Am. J. Physiol. Heart Circ. Physiol. 296, H2007–H2014 (2009).
Charo, D.N. et al. Endogenous regulation of cardiovascular function by apelin-APJ. Am. J. Physiol. Heart Circ. Physiol. 297, H1904–H1913 (2009).
Habata, Y. et al. Apelin, the natural ligand of the orphan receptor APJ, is abundantly secreted in the colostrum. Biochim. Biophys. Acta 1452, 25–35 (1999).
Chamorro-Jorganes, A. et al. MicroRNA-16 and microRNA-424 regulate cell-autonomous angiogenic functions in endothelial cells via targeting vascular endothelial growth factor receptor-2 and fibroblast growth factor receptor-1. Arterioscler. Thromb. Vasc. Biol. 31, 2595–2606 (2011).
Murakami, M. et al. The FGF system has a key role in regulating vascular integrity. J. Clin. Invest. 118, 3355–3366 (2008).
Stenmark, K.R., Meyrick, B., Galie, N., Mooi, W.J. & McMurtry, I.F. Animal models of pulmonary arterial hypertension: the hope for etiological discovery and pharmacological cure. Am. J. Physiol. Lung Cell. Mol. Physiol. 297, L1013–L1032 (2009).
Taraseviciene-Stewart, L. et al. Inhibition of the VEGF receptor 2 combined with chronic hypoxia causes cell death–dependent pulmonary endothelial cell proliferation and severe pulmonary hypertension. FASEB J. 15, 427–438 (2001).
Tu, L. et al. Autocrine fibroblast growth factor-2 signaling contributes to altered endothelial phenotype in pulmonary hypertension. Am. J. Respir. Cell Mol. Biol. 45, 311–322 (2011).
Masri, B., Morin, N., Cornu, M., Knibiehler, B. & Audigier, Y. Apelin (65–77) activates p70 S6 kinase and is mitogenic for umbilical endothelial cells. FASEB J. 18, 1909–1911 (2004).
Eyries, M. et al. Hypoxia-induced apelin expression regulates endothelial cell proliferation and regenerative angiogenesis. Circ. Res. 103, 432–440 (2008).
Kälin, R.E. et al. Paracrine and autocrine mechanisms of apelin signaling govern embryonic and tumor angiogenesis. Dev. Biol. 305, 599–614 (2007).
del Toro, R. et al. Identification and functional analysis of endothelial tip cell-enriched genes. Blood 116, 4025–4033 (2010).
Kasai, A. et al. Apelin is a crucial factor for hypoxia-induced retinal angiogenesis. Arterioscler. Thromb. Vasc. Biol. 30, 2182–2187 (2010).
Kasai, A. et al. Retardation of retinal vascular development in apelin-deficient mice. Arterioscler. Thromb. Vasc. Biol. 28, 1717–1722 (2008).
Kidoya, H., Naito, H. & Takakura, N. Apelin induces enlarged and nonleaky blood vessels for functional recovery from ischemia. Blood 115, 3166–3174 (2010).
Caruso, P. et al. Dynamic changes in lung microRNA profiles during the development of pulmonary hypertension due to chronic hypoxia and monocrotaline. Arterioscler. Thromb. Vasc. Biol. 30, 716–723 (2010).
Forrest, A.R. et al. Induction of microRNAs, mir-155, mir-222, mir-424 and mir-503, promotes monocytic differentiation through combinatorial regulation. Leukemia 24, 460–466 (2010).
Sarkar, S., Dey, B.K. & Dutta, A. MiR-322/424 and -503 are induced during muscle differentiation and promote cell cycle quiescence and differentiation by down-regulation of Cdc25A. Mol. Biol. Cell 21, 2138–2149 (2010).
Ghosh, G. et al. Hypoxia-induced microRNA-424 expression in human endothelial cells regulates HIF-alpha isoforms and promotes angiogenesis. J. Clin. Invest. 120, 4141–4154 (2010).
Comhair, S.A. et al. Human primary lung endothelial cells in culture. Am. J. Respir. Cell Mol. Biol. 46, 723–730 (2012).
Aytekin, M. et al. High levels of hyaluronan in idiopathic pulmonary arterial hypertension. Am. J. Physiol. Lung Cell. Mol. Physiol. 295, L789–L799 (2008).
Chun, H.J. et al. Apelin signaling antagonizes Ang II effects in mouse models of atherosclerosis. J. Clin. Invest. 118, 3343–3354 (2008).
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.).
Author information
Authors and Affiliations
Contributions
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.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–19 and Supplementary Tables 1 and 2 (PDF 1819 kb)
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nm.3040
This article is cited by
-
Comprehensive analysis of the diagnostic and therapeutic value of the hypoxia-related gene PLAUR in the progression of atherosclerosis
Scientific Reports (2023)
-
Growth differentiation factor 11 induces skeletal muscle atrophy via a STAT3-dependent mechanism in pulmonary arterial hypertension
Skeletal Muscle (2022)
-
Upregulated anti-angiogenic miR-424-5p in type 1 diabetes (model of subclinical cardiovascular disease) correlates with endothelial progenitor cells, CXCR1/2 and other parameters of vascular health
Stem Cell Research & Therapy (2021)
-
Identifying metastasis-initiating miRNA-target regulations of colorectal cancer from expressional changes in primary tumors
Scientific Reports (2020)
-
Therapeutic effects of histone deacetylase inhibitors on heart disease
Archives of Pharmacal Research (2020)