Abstract
Pulmonary hypertension (PH) is characterized by increased proliferation and apoptosis resistance of pulmonary artery smooth muscle cells (PASMCs). Forkhead box O (FoxO) transcription factors are key regulators of cellular proliferation. Here we show that in pulmonary vessels and PASMCs of human and experimental PH lungs, FoxO1 expression is downregulated and FoxO1 is inactivated via phosphorylation and nuclear exclusion. These findings could be reproduced using ex vivo exposure of PASMCs to growth factors and inflammatory cytokines. Pharmacological inhibition and genetic ablation of FoxO1 in smooth muscle cells reproduced PH features in vitro and in vivo. Either pharmacological reconstitution of FoxO1 activity using intravenous or inhaled paclitaxel, or reconstitution of the transcriptional activity of FoxO1 by gene therapy, restored the physiologically quiescent PASMC phenotype in vitro, linked to changes in cell cycle control and bone morphogenic protein receptor type 2 (BMPR2) signaling, and reversed vascular remodeling and right-heart hypertrophy in vivo. Thus, PASMC FoxO1 is a critical integrator of multiple signaling pathways driving PH, and reconstitution of FoxO1 activity offers a potential therapeutic option for PH.
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Acknowledgements
We thank J. Clardy and R. Kalb from Harvard Medical School for kindly providing Sam A. We thank M. Rabinovitch (Stanford University School of Medicine) for providing the mouse PASMC isolation protocol. We also acknowledge the excellent technical assistance of K. Leib in performing cell culture experiments, U. Eule, C. Vroom and A. Hecker in intact animal experiments and M. Schmoranzer, M. Hoeck and V. Golchert in histological studies. We thank K. Quanz for performing isolated lung experiments. We acknowledge M. Potente for providing genotyping protocol and siRNAs. This work was supported by the Max Planck Society, the Scientific and Economic Excellence in Hesse (LOEWE) Programm and the German Research Foundation through Excellence Cluster 147 Cardio-Pulmonary System (ECCPS), Cluster of Excellence REBIRTH (From Regenerative Biology to Reconstructive Therapy).
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R.S., D.S., H.M.A.-T., B.K., C.M., R.T.S. and S.S.P. acquired the data. R.S., D.S., H.M.A.-T., B.K., C.M., N.W., R.T.S. and S.S.P. analyzed and interpreted the data. R.S., N.W., R.T.S., D.S., W.S. and S.S.P. conceived and designed research. M.R.C. contributed transgenic mice. R.S., W.S., F.G. and S.S.P. drafted the manuscript. R.S., W.S., N.W., R.T.S. and S.S.P. handled the funding and supervision.
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Savai, R., Al-Tamari, H., Sedding, D. et al. Pro-proliferative and inflammatory signaling converge on FoxO1 transcription factor in pulmonary hypertension. Nat Med 20, 1289–1300 (2014). https://doi.org/10.1038/nm.3695
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DOI: https://doi.org/10.1038/nm.3695
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