TGF‐β signaling is dynamically regulated during the alveolarization of rodent and human lungs

Alejandre-Alcázar, Miguel A.; Michiels-Corsten, Matthias; Vicencio, Alfin G.; Ryu, Julie; Krijger, Ronald R. de; Haddad, Gabriel G.; Tibboel, Dick; Seeger, Werner; Eickelberg, Oliver; Morty, Rory E.

doi:10.1002/dvdy.21403

Cited by 90 publications

(59 citation statements)

References 58 publications

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“…This protein is present in pulmonary vascular tissue of fetal, premature, and term neonates (1,10), and its expression increases in infants developing chronic lung disease (7). Given its known angiogenic effects (19), its presence early in life and enhanced expression in normal lungs suggest that endoglin is important for pulmonary vascular development and branching.…”

Section: Discussionmentioning

confidence: 99%

Age-dependent endothelial nitric oxide synthase uncoupling in pulmonary arteries of endoglin heterozygous mice

Belik

Jerkić

McIntyre³

et al. 2009

American Journal of Physiology-Lung Cellular and Molecular Phys

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, and clinical signs of disease are generally more evident later in life. We previously showed that systemic vessels of adult Eng heterozygous (Eng ϩ/Ϫ ) mice exhibit increased vasorelaxation due to uncoupling of endothelial nitric oxide synthase (eNOS). We postulated that these changes may develop with age and evaluated pulmonary arteries from newborn and adult Eng ϩ/Ϫ mice for eNOS-dependent, acetylcholine (ACh-induced) vasorelaxation, compared with that of age-matched littermate controls. While ACh-induced vasorelaxation was similar in all newborn mice, it was significantly increased in the adult Eng ϩ/Ϫ vs. control vessels. The vasodilatory responses were inhibited by L-NAME suggesting eNOS dependence. eNOS uncoupling was observed in lung tissues of adult, but not newborn, heterozygous mice and was associated with increased production of reactive O2 species (ROS) in adult Eng ϩ/Ϫ vs. control lungs. Interestingly, ROS generation was higher in adult than newborn mice and so were the levels of NADPH oxidase 4 and SOD 1, 2, 3 isoforms. However, enzyme protein levels and NADPH activity were normal in adult Eng ϩ/Ϫ lungs indicating that the developmental maturation of ROS generation and scavenging cannot account for the increased vasodilatation observed in adult Eng ϩ/Ϫ mice. Our data suggest that eNOS-dependent H2O2 generation in Eng ϩ/Ϫ lungs accounts for the heightened pulmonary vasorelaxation. To the extent that these mice mimic human HHT1, age-associated pulmonary vascular eNOS uncoupling may explain the late childhood and adult onset of clinical lung manifestations. lung; newborn; pulmonary vascular resistance; hereditary hemorrhagic telangiectasia LUNG BLOOD FLOW IS MOSTLY dependent on the regional arteriolar and venular intraluminal diameter that ultimately determines the pulmonary vascular resistance (PVR). Nitric oxide (NO) is constitutively produced by endothelial and smooth muscle cells (39) via synthases (NOS) of which there are three isoforms: endothelial (eNOS), neuronal (nNOS), and inducible (iNOS). Although there is evidence that iNOS and nNOS are expressed in the fetal pulmonary vasculature (38), lung eNOS protein expression increases during gestation suggesting that its vascular tissue content and/or activity is in part responsible for the high PVR prenatally and the changes occurring after birth (15). In sheep, lung eNOS expression is maximal in late gestation (34), whereas in rats it is highest either before (33), or immediately after, birth (23). Postnatally, lung vascular tissue eNOS expression was shown to decrease with age in pigs (17).eNOS converts L-arginine to L-citrulline to generate NO. Its activity is dependent on Ca 2ϩ /calmodulin (CaM), but also on subcellular localization, posttranslational modifications, and interaction with several regulatory proteins, including Hsp90 (11,14). Hsp90 facilitates CaM-induced release from caveolae and acts as a scaffold factor for eNOS; it is necessary for eNOS phosphorylation at Ser1177 (13, 43). This complex process controls the state of eNOS ac...

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Section: Discussionmentioning

confidence: 99%

Age-dependent endothelial nitric oxide synthase uncoupling in pulmonary arteries of endoglin heterozygous mice

Belik

Jerkić

McIntyre³

et al. 2009

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…2B), suggesting a role for Smad1, which is the key mediator of the Acvrl1/Smad1 axis, in the effects of dexamethasone on TGF-␤ signaling. The Acvrl1/Smad1 axis, which is traditionally considered to be active primarily in the endothelium (29, 48 -50), was demonstrated to be active in NIH/3T3 cells (results not shown) and in fibroblasts (51,52). To examine the functional contribution of Smad1 to the effects of dexamethasone on TGF-␤ signaling, the expression of smad1 was ablated by transfection of NIH/3T3 cells with siRNA directed against smad1, with scrambled siRNA serving as a negative control.…”

Section: Glucocorticoids Inhibit Classical Tgf-␤ Signaling-mentioning

confidence: 90%

Glucocorticoids Recruit Tgfbr3 and Smad1 to Shift Transforming Growth Factor-β Signaling from the Tgfbr1/Smad2/3 Axis to the Acvrl1/Smad1 Axis in Lung Fibroblasts

Schwartze

Becker

Sakkas

et al. 2014

Journal of Biological Chemistry

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“…Real-time RT-PCR was undertaken exactly as described previously [6, 11, 12], using primers listed in table S1, all of which generated C t values below 35 (fig. S2), for human (see table 1 for number of subjects per group), mouse ( n = 5, per group) or cell culture ( n = 3 per group) material.…”

Section: Methodsmentioning

confidence: 99%

“…S2), for human (see table 1 for number of subjects per group), mouse ( n = 5, per group) or cell culture ( n = 3 per group) material. The cDNA was synthesized as described previously [6, 11, 12] from total RNA pools prepared from lung tissue or cultured cell homogenates. Immunoblotting was performed exactly as previously described [6, 11, 12] using: goat anti-Tgm1 (SantaCruz, SC-18127; 1:200); goat anti-Tgm2 (Upstate, 06-471; 1:1000); and rabbit anti-α-tubulin (SantaCruz, SC-5286, 1:2500).…”

Section: Methodsmentioning

confidence: 99%

“…The cDNA was synthesized as described previously [6, 11, 12] from total RNA pools prepared from lung tissue or cultured cell homogenates. Immunoblotting was performed exactly as previously described [6, 11, 12] using: goat anti-Tgm1 (SantaCruz, SC-18127; 1:200); goat anti-Tgm2 (Upstate, 06-471; 1:1000); and rabbit anti-α-tubulin (SantaCruz, SC-5286, 1:2500). Immune complexes were detected ( n = 3 per group, per time-point) with donkey anti-goat IgG horseradish peroxidase conjugate (SantaCruz, SC-2020; 1:1000) and goat anti-rabbit IgG horseradish peroxidase conjugate (Pierce, 31460; 1:3000), using chemiluminescence.…”

Section: Methodsmentioning

confidence: 99%

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Transglutaminase 2: a new player in bronchopulmonary dysplasia?

et al. 2014

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Aberrant remodelling of the extracellular matrix (ECM) in the developing lung may underlie arrested alveolarisation associated with bronchopulmonary dysplasia (BPD). Transglutaminases are regulators of ECM remodelling. Therefore, the expression and activity of transglutaminases were assessed in lungs from human neonates with BPD, and in a rodent model of BPD. Transglutaminase expression and localisation were assessed by RT-PCR, immunoblot, activity assay, and immunohistochemical analyses of human and mouse lung tissues. Transglutaminase regulation by transforming growth factor (TGF)-β was investigated in lung cells by luciferase-based reporter assay, and RT-PCR. TGF-β signalling was neutralised in vivo in an animal model of BPD, to determine whether TGF-β mediated the hyperoxia-induced changes in transglutaminase expression. Transglutaminase 2 expression was upregulated in the lungs of preterm infants with BPD, and in the lungs of hyperoxia-exposed mouse pups, where lung development was arrested. Transglutaminase 2 localised to the developing alveolar septa. TGF-β was identified as a regulator of transglutaminase 2 expression in human and mouse lung epithelial cells. In vivo neutralisation of TGF-β signalling partially restored normal lung structure and normalised lung transglutaminase 2 mRNA expression. Our data point to a role for perturbed transglutaminase 2 activity in the arrested alveolarisation associated with BPD.

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TGF‐β signaling is dynamically regulated during the alveolarization of rodent and human lungs

Cited by 90 publications

References 58 publications

Age-dependent endothelial nitric oxide synthase uncoupling in pulmonary arteries of endoglin heterozygous mice

Age-dependent endothelial nitric oxide synthase uncoupling in pulmonary arteries of endoglin heterozygous mice

Glucocorticoids Recruit Tgfbr3 and Smad1 to Shift Transforming Growth Factor-β Signaling from the Tgfbr1/Smad2/3 Axis to the Acvrl1/Smad1 Axis in Lung Fibroblasts

Transglutaminase 2: a new player in bronchopulmonary dysplasia?

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