17β-Estradiol and estrogen receptor α protect right ventricular function in pulmonary hypertension via BMPR2 and apelin
Andrea L. Frump, … , Sebastien Bonnet, Tim Lahm
Andrea L. Frump, … , Sebastien Bonnet, Tim Lahm
Published January 26, 2021
Citation Information: J Clin Invest. 2021;131(6):e129433. https://doi.org/10.1172/JCI129433.
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Research Article Pulmonology Vascular biology

17β-Estradiol and estrogen receptor α protect right ventricular function in pulmonary hypertension via BMPR2 and apelin

Abstract

Women with pulmonary arterial hypertension (PAH) exhibit better right ventricular (RV) function and survival than men; however, the underlying mechanisms are unknown. We hypothesized that 17β-estradiol (E2), through estrogen receptor α (ER-α), attenuates PAH-induced RV failure (RVF) by upregulating the procontractile and prosurvival peptide apelin via a BMPR2-dependent mechanism. We found that ER-α and apelin expression were decreased in RV homogenates from patients with RVF and from rats with maladaptive (but not adaptive) RV remodeling. RV cardiomyocyte apelin abundance increased in vivo or in vitro after treatment with E2 or ER-α agonist. Studies employing ER-α–null or ER-β–null mice, ER-α loss-of-function mutant rats, or siRNA demonstrated that ER-α is necessary for E2 to upregulate RV apelin. E2 and ER-α increased BMPR2 in pulmonary hypertension RVs and in isolated RV cardiomyocytes, associated with ER-α binding to the Bmpr2 promoter. BMPR2 is required for E2-mediated increases in apelin abundance, and both BMPR2 and apelin are necessary for E2 to exert RV-protective effects. E2 or ER-α agonist rescued monocrotaline pulmonary hypertension and restored RV apelin and BMPR2. We identified what we believe to be a novel cardioprotective E2/ER-α/BMPR2/apelin axis in the RV. Harnessing this axis may lead to novel RV-targeted therapies for PAH patients of either sex.

Authors

Andrea L. Frump, Marjorie Albrecht, Bakhtiyor Yakubov, Sandra Breuils-Bonnet, Valérie Nadeau, Eve Tremblay, Francois Potus, Junichi Omura, Todd Cook, Amanda Fisher, Brooke Rodriguez, R. Dale Brown, Kurt R. Stenmark, C. Dustin Rubinstein, Kathy Krentz, Diana M. Tabima, Rongbo Li, Xin Sun, Naomi C. Chesler, Steeve Provencher, Sebastien Bonnet, Tim Lahm

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Figure 10

ER-α binds to Bmpr2 promoter and is necessary and sufficient to increase RV BMPR2 in vitro and in vivo.

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ER-α binds to Bmpr2 promoter and is necessary and sufficient to increase...
(A) RV BMPR2 and ER-α protein correlate positively in male and female control and SuHx-PH rats. (B) ChIP of ER-α binding at the Bmpr2 promoter. H9c2 cardiomyoblasts were treated with E2 (100 nM) for 0 (control), 4, or 20 hours. DNA/protein complexes were cross-linked and immunoprecipitated with anti–ER-α antibody or IgG isotype control. RNA polymerase II (RNA Pol II) binding to Gapdh promoter was used as positive control (bottom panel). NTC: no template control. (C) Time course of BMPR2 expression in H9c2 cardiomyoblasts after ER-α siRNA knockdown (5 nM; 24 hours prior to E2 [100 nM]; see Figure 4B for ER-α knockdown efficacy). (D) BMPR2 protein in H9c2 cells treated with ER-α agonist BTP-α (100 nM, 24 hours). (E) BMPR2 protein in RV cardiomyocytes (RVCMs) isolated from male rats and treated with E2 (10 nM, 24 hours) or BTP-α (100 nM, 24 hours) in vitro. (F) BMPR2 protein in RVCMs isolated from male and female SuHx-PH rats and treated in vitro with BTP-α (100 nM, 24 hours). (G) BMPR2 expression in RV homogenates from male normoxia, SuHx-PH, or SuHx-PH rats treated with E2 or ER-α agonist PPT (75 or 850 μg/kg/day; s.c. pellets). (H) BMPR2 protein in RVCMs from groups shown in F. n = 3 independent experiments in B–D; n = 4 rats/group in E–H. B–G depict representative Western blots. Densitometries include data from all experiments or animals. Pearson’s R value and P value shown in A. Dashed lines represent 95% CI. *P < 0.05 versus Scr (scrambled control), $P < 0.05 versus E2 in C; *P < 0.05 versus control or normoxia in D–H; #P < 0.05 versus untreated SuHx in F and H. P values in B, C, and E–H by ANOVA/post hoc Tukey correction; P value in D by Student’s t test. Error bars represent mean ± SEM; each data point represents 1 experiment/animal.