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Selective enhancement of endothelial BMPR-II with BMP9 reverses pulmonary arterial hypertension.

Long L, Ormiston ML, Yang X, Southwood M, Gräf S, Machado RD, Mueller M, Kinzel B, Yung LM, Wilkinson JM, Moore SD, Drake KM, Aldred MA, Yu PB, Upton PD, Morrell NW - Nat. Med. (2015)

Bottom Line: However, selective targeting of this signaling pathway using BMP ligands has not yet been explored as a therapeutic strategy.Administration of BMP9 reversed established PAH in these mice, as well as in two other experimental PAH models, in which PAH develops in response to either monocrotaline or VEGF receptor inhibition combined with chronic hypoxia.These results demonstrate the promise of direct enhancement of endothelial BMP signaling as a new therapeutic strategy for PAH.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK.

ABSTRACT
Genetic evidence implicates the loss of bone morphogenetic protein type II receptor (BMPR-II) signaling in the endothelium as an initiating factor in pulmonary arterial hypertension (PAH). However, selective targeting of this signaling pathway using BMP ligands has not yet been explored as a therapeutic strategy. Here, we identify BMP9 as the preferred ligand for preventing apoptosis and enhancing monolayer integrity in both pulmonary arterial endothelial cells and blood outgrowth endothelial cells from subjects with PAH who bear mutations in the gene encoding BMPR-II, BMPR2. Mice bearing a heterozygous knock-in allele of a human BMPR2 mutation, R899X, which we generated as an animal model of PAH caused by BMPR-II deficiency, spontaneously developed PAH. Administration of BMP9 reversed established PAH in these mice, as well as in two other experimental PAH models, in which PAH develops in response to either monocrotaline or VEGF receptor inhibition combined with chronic hypoxia. These results demonstrate the promise of direct enhancement of endothelial BMP signaling as a new therapeutic strategy for PAH.

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BMP9 reverses established pulmonary hypertension in rats in response to treatment with the VEGF receptor inhibitior SU-5416 in combination with chronic hypoxia(a) Rats were given vehicle injections and maintained in normoxia (n=4) or challenged with SU-5416 (20 mg/kg, i.p.) and 3 weeks hypoxia (10% O2) prior to 5 weeks normoxia and assessment at 8 weeks (n=7) or at 11 weeks following daily treatment with saline vehicle (n=11) or BMP9 (n=11; 600 ng/day, i.p.). (b) Assessment of right ventricular systolic pressure (RVSP) and (c) RV hypertrophy (Fulton index) for the rats described in a (1-way ANOVA, Tukey’s post test). (d) Quantification of non-, partially- and fully-muscularized arteries as a percentage of total alveolar wall and duct arteries (n=4 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test for fully muscularized vessels). (e) Assessment of pulmonary arterial wall thickness as a percentage of luminal diameter (n=4 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). (f) Quantification of neointimal lesion frequency in the lungs of the rats described in a (n=3 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). (g) Quantification of cleaved caspase-3 positive endothelial cells in lung sections from the groups described in a (n=3 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). ***P<0.001, **P<0.01, *P<0.05. Mean +/− SEM. (h) Neointima formation in the lungs of the rats described in a. Lung sections were stained for smooth muscle α-actin (SMA) or with elastic van Gieson (EVG) stain. Scale bar = 50μm.
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Figure 6: BMP9 reverses established pulmonary hypertension in rats in response to treatment with the VEGF receptor inhibitior SU-5416 in combination with chronic hypoxia(a) Rats were given vehicle injections and maintained in normoxia (n=4) or challenged with SU-5416 (20 mg/kg, i.p.) and 3 weeks hypoxia (10% O2) prior to 5 weeks normoxia and assessment at 8 weeks (n=7) or at 11 weeks following daily treatment with saline vehicle (n=11) or BMP9 (n=11; 600 ng/day, i.p.). (b) Assessment of right ventricular systolic pressure (RVSP) and (c) RV hypertrophy (Fulton index) for the rats described in a (1-way ANOVA, Tukey’s post test). (d) Quantification of non-, partially- and fully-muscularized arteries as a percentage of total alveolar wall and duct arteries (n=4 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test for fully muscularized vessels). (e) Assessment of pulmonary arterial wall thickness as a percentage of luminal diameter (n=4 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). (f) Quantification of neointimal lesion frequency in the lungs of the rats described in a (n=3 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). (g) Quantification of cleaved caspase-3 positive endothelial cells in lung sections from the groups described in a (n=3 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). ***P<0.001, **P<0.01, *P<0.05. Mean +/− SEM. (h) Neointima formation in the lungs of the rats described in a. Lung sections were stained for smooth muscle α-actin (SMA) or with elastic van Gieson (EVG) stain. Scale bar = 50μm.

Mentions: We also examined the efficacy of BMP9 in the more severe PAH model of rats exposed to chronic hypoxia in combination with the VEGF receptor blocker SU-541617 (Fig. 6a). Rats given SU-5416 and exposed to three weeks of hypoxia (10% O2) and an additional 5 weeks of normoxia developed severe pulmonary hypertension and right ventricular hypertrophy (Fig. 6b–c) as well as extensive pulmonary arterial muscularization (Fig. 6d–e). Daily BMP9 (600 ng/day, i.p.) from weeks 8 to 11 resulted in a reversal of established disease, including a reduction in RVSP, RV hypertrophy and vascular remodeling when compared to saline-treated controls (Fig. 6b–e).


Selective enhancement of endothelial BMPR-II with BMP9 reverses pulmonary arterial hypertension.

Long L, Ormiston ML, Yang X, Southwood M, Gräf S, Machado RD, Mueller M, Kinzel B, Yung LM, Wilkinson JM, Moore SD, Drake KM, Aldred MA, Yu PB, Upton PD, Morrell NW - Nat. Med. (2015)

BMP9 reverses established pulmonary hypertension in rats in response to treatment with the VEGF receptor inhibitior SU-5416 in combination with chronic hypoxia(a) Rats were given vehicle injections and maintained in normoxia (n=4) or challenged with SU-5416 (20 mg/kg, i.p.) and 3 weeks hypoxia (10% O2) prior to 5 weeks normoxia and assessment at 8 weeks (n=7) or at 11 weeks following daily treatment with saline vehicle (n=11) or BMP9 (n=11; 600 ng/day, i.p.). (b) Assessment of right ventricular systolic pressure (RVSP) and (c) RV hypertrophy (Fulton index) for the rats described in a (1-way ANOVA, Tukey’s post test). (d) Quantification of non-, partially- and fully-muscularized arteries as a percentage of total alveolar wall and duct arteries (n=4 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test for fully muscularized vessels). (e) Assessment of pulmonary arterial wall thickness as a percentage of luminal diameter (n=4 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). (f) Quantification of neointimal lesion frequency in the lungs of the rats described in a (n=3 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). (g) Quantification of cleaved caspase-3 positive endothelial cells in lung sections from the groups described in a (n=3 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). ***P<0.001, **P<0.01, *P<0.05. Mean +/− SEM. (h) Neointima formation in the lungs of the rats described in a. Lung sections were stained for smooth muscle α-actin (SMA) or with elastic van Gieson (EVG) stain. Scale bar = 50μm.
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Figure 6: BMP9 reverses established pulmonary hypertension in rats in response to treatment with the VEGF receptor inhibitior SU-5416 in combination with chronic hypoxia(a) Rats were given vehicle injections and maintained in normoxia (n=4) or challenged with SU-5416 (20 mg/kg, i.p.) and 3 weeks hypoxia (10% O2) prior to 5 weeks normoxia and assessment at 8 weeks (n=7) or at 11 weeks following daily treatment with saline vehicle (n=11) or BMP9 (n=11; 600 ng/day, i.p.). (b) Assessment of right ventricular systolic pressure (RVSP) and (c) RV hypertrophy (Fulton index) for the rats described in a (1-way ANOVA, Tukey’s post test). (d) Quantification of non-, partially- and fully-muscularized arteries as a percentage of total alveolar wall and duct arteries (n=4 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test for fully muscularized vessels). (e) Assessment of pulmonary arterial wall thickness as a percentage of luminal diameter (n=4 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). (f) Quantification of neointimal lesion frequency in the lungs of the rats described in a (n=3 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). (g) Quantification of cleaved caspase-3 positive endothelial cells in lung sections from the groups described in a (n=3 for control, n=6 for all other groups; 1-way ANOVA, Tukey’s post test). ***P<0.001, **P<0.01, *P<0.05. Mean +/− SEM. (h) Neointima formation in the lungs of the rats described in a. Lung sections were stained for smooth muscle α-actin (SMA) or with elastic van Gieson (EVG) stain. Scale bar = 50μm.
Mentions: We also examined the efficacy of BMP9 in the more severe PAH model of rats exposed to chronic hypoxia in combination with the VEGF receptor blocker SU-541617 (Fig. 6a). Rats given SU-5416 and exposed to three weeks of hypoxia (10% O2) and an additional 5 weeks of normoxia developed severe pulmonary hypertension and right ventricular hypertrophy (Fig. 6b–c) as well as extensive pulmonary arterial muscularization (Fig. 6d–e). Daily BMP9 (600 ng/day, i.p.) from weeks 8 to 11 resulted in a reversal of established disease, including a reduction in RVSP, RV hypertrophy and vascular remodeling when compared to saline-treated controls (Fig. 6b–e).

Bottom Line: However, selective targeting of this signaling pathway using BMP ligands has not yet been explored as a therapeutic strategy.Administration of BMP9 reversed established PAH in these mice, as well as in two other experimental PAH models, in which PAH develops in response to either monocrotaline or VEGF receptor inhibition combined with chronic hypoxia.These results demonstrate the promise of direct enhancement of endothelial BMP signaling as a new therapeutic strategy for PAH.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK.

ABSTRACT
Genetic evidence implicates the loss of bone morphogenetic protein type II receptor (BMPR-II) signaling in the endothelium as an initiating factor in pulmonary arterial hypertension (PAH). However, selective targeting of this signaling pathway using BMP ligands has not yet been explored as a therapeutic strategy. Here, we identify BMP9 as the preferred ligand for preventing apoptosis and enhancing monolayer integrity in both pulmonary arterial endothelial cells and blood outgrowth endothelial cells from subjects with PAH who bear mutations in the gene encoding BMPR-II, BMPR2. Mice bearing a heterozygous knock-in allele of a human BMPR2 mutation, R899X, which we generated as an animal model of PAH caused by BMPR-II deficiency, spontaneously developed PAH. Administration of BMP9 reversed established PAH in these mice, as well as in two other experimental PAH models, in which PAH develops in response to either monocrotaline or VEGF receptor inhibition combined with chronic hypoxia. These results demonstrate the promise of direct enhancement of endothelial BMP signaling as a new therapeutic strategy for PAH.

Show MeSH
Related in: MedlinePlus