FK506 activates BMPR2, rescues endothelial dysfunction, and reverses pulmonary hypertension
Edda Spiekerkoetter, … , Peter ten Dijke, Marlene Rabinovitch
Edda Spiekerkoetter, … , Peter ten Dijke, Marlene Rabinovitch
Published July 15, 2013
Citation Information: J Clin Invest. 2013;123(8):3600-3613. https://doi.org/10.1172/JCI65592.
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Research Article Vascular biology

FK506 activates BMPR2, rescues endothelial dysfunction, and reverses pulmonary hypertension

Abstract

Dysfunctional bone morphogenetic protein receptor-2 (BMPR2) signaling is implicated in the pathogenesis of pulmonary arterial hypertension (PAH). We used a transcriptional high-throughput luciferase reporter assay to screen 3,756 FDA-approved drugs and bioactive compounds for induction of BMPR2 signaling. The best response was achieved with FK506 (tacrolimus), via a dual mechanism of action as a calcineurin inhibitor that also binds FK-binding protein-12 (FKBP12), a repressor of BMP signaling. FK506 released FKBP12 from type I receptors activin receptor-like kinase 1 (ALK1), ALK2, and ALK3 and activated downstream SMAD1/5 and MAPK signaling and ID1 gene regulation in a manner superior to the calcineurin inhibitor cyclosporine and the FKBP12 ligand rapamycin. In pulmonary artery endothelial cells (ECs) from patients with idiopathic PAH, low-dose FK506 reversed dysfunctional BMPR2 signaling. In mice with conditional Bmpr2 deletion in ECs, low-dose FK506 prevented exaggerated chronic hypoxic PAH associated with induction of EC targets of BMP signaling, such as apelin. Low-dose FK506 also reversed severe PAH in rats with medial hypertrophy following monocrotaline and in rats with neointima formation following VEGF receptor blockade and chronic hypoxia. Our studies indicate that low-dose FK506 could be useful in the treatment of PAH.

Authors

Edda Spiekerkoetter, Xuefei Tian, Jie Cai, Rachel K. Hopper, Deepti Sudheendra, Caiyun G. Li, Nesrine El-Bizri, Hirofumi Sawada, Roxanna Haghighat, Roshelle Chan, Leila Haghighat, Vinicio de Jesus Perez, Lingli Wang, Sushma Reddy, Mingming Zhao, Daniel Bernstein, David E. Solow-Cordero, Philip A. Beachy, Thomas J. Wandless, Peter ten Dijke, Marlene Rabinovitch

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Figure 6

Low-dose FK506 reverses established PAH and neointima formation in rats.

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Low-dose FK506 reverses established PAH and neointima formation in rats....
(A) Timeline of experiments. (B) RVSP and RVH (Fulton index) in rats in normoxia with or without vehicle (VEH) or FK506 and in rats after SUGEN with 3 weeks hypoxia and 5 weeks normoxia, sacrificed at 8 weeks and at 11 weeks following treatment with vehicle or FK506 (n = 8; *P < 0.05; ***P < 0.001 vs. same condition in normoxia, 2-way ANOVA, Bonferroni post test). (C) Representative images (scale bar: 100 μm) and percentage vessels with neointima in hypoxia/SUGEN/normoxia rats at 8 and 11 weeks treated with vehicle or FK506 (n = 8; **P < 0.01 vs. vehicle, 1-way ANOVA, Bonferroni post test). (D) Number of alveoli per mm2 in normoxia (with or without vehicle or FK506) and in rats after hypoxia/SUGEN/normoxia, sacrificed at 11 weeks (with or without 3-week treatment of vehicle or FK506) (n = 8; *P < 0.05 vs. untreated normoxia condition, 2-way ANOVA, Dunnett’s post test). (E) Mean number of arteries per 100 alveoli (5 fields ×20) in same groups as in D (n = 8, #P < 0.05, 1-way ANOVA, selected pairs post test, vehicle vs. FK506). (F) Apelin/β-actin mRNA expression in total lung lysate in same group as in D (n = 8; *P < 0.05, 1-way ANOVA, Dunnett’s post test). (G) Il2/Gapdh mRNA expression in whole lungs of rats in normoxia (with or without vehicle or FK506) and in rats after hypoxia/SUGEN/normoxia, sacrificed at 11 weeks (with or without 3-week treatment of vehicle or FK506) (n = 8; *P < 0.05 vs. vehicle normoxia, #P < 0.05 vs. FK normoxia, nonsignificant trend in decrease of IL-2 with FK506 treatment, 2-way ANOVA, Bonferroni multiple-comparison test). Mean ± SEM.