Sequential Ligand-Dependent Notch Signaling Activation Regulates Valve Primordium Formation and Morphogenesis

Donal MacGrogan, Gaetano D'Amato, Stanislao Travisano, Beatriz Martinez-Poveda, Guillermo Luxán, Gonzalo Del Monte-Nieto, Tania Papoutsi, Mauro Sbroggio, Vanesa Bou, Pablo Gomez-Del Arco, Manuel Jose Gómez, Bin Zhou, Juan Miguel Redondo, Luis J. Jiménez-Borreguero, José Luis De La Pompa

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Rationale: The Notch signaling pathway is crucial for primitive cardiac valve formation by epithelial-mesenchymal transition, and NOTCH1 mutations cause bicuspid aortic valve; however, the temporal requirement for the various Notch ligands and receptors during valve ontogeny is poorly understood. Objective: The aim of this study is to determine the functional specificity of Notch in valve development. Methods and Results: Using cardiac-specific conditional targeted mutant mice, we find that endothelial/endocardial deletion of Mib1-Dll4-Notch1 signaling, possibly favored by Manic-Fringe, is specifically required for cardiac epithelial-mesenchymal transition. Mice lacking endocardial Jag1, Notch1, or RBPJ displayed enlarged valve cusps, bicuspid aortic valve, and septal defects, indicating that endocardial Jag1 to Notch1 signaling is required for post-epithelial-mesenchymal transition valvulogenesis. Valve dysmorphology was associated with increased mesenchyme proliferation, indicating that Jag1-Notch1 signaling restricts mesenchyme cell proliferation non-cell autonomously. Gene profiling revealed upregulated Bmp signaling in Jag1-mutant valves, providing a molecular basis for the hyperproliferative phenotype. Significantly, the negative regulator of mesenchyme proliferation, Hbegf, was markedly reduced in Jag1-mutant valves. Hbegf expression in embryonic endocardial cells could be readily activated through a RBPJ-binding site, identifying Hbegf as an endocardial Notch target. Accordingly, addition of soluble heparin-binding EGF-like growth factor to Jag1-mutant outflow tract explant cultures rescued the hyperproliferative phenotype. Conclusions: During cardiac valve formation, Dll4-Notch1 signaling leads to epithelial-mesenchymal transition and cushion formation. Jag1-Notch1 signaling subsequently restrains Bmp-mediated valve mesenchyme proliferation by sustaining Hbegf-EGF receptor signaling. Our studies identify a mechanism of signaling cross talk during valve morphogenesis involved in the origin of congenital heart defects associated with reduced NOTCH function.

Original languageEnglish (US)
Pages (from-to)1480-1497
Number of pages18
JournalCirculation Research
Volume118
Issue number10
DOIs
StatePublished - May 13 2016

Fingerprint

Epithelial-Mesenchymal Transition
Mesoderm
Morphogenesis
Ligands
Heart Valves
Notch Receptors
Phenotype
Congenital Heart Defects
Epidermal Growth Factor Receptor
Binding Sites
Cell Proliferation
Mutation
Genes
Bicuspid Aortic Valve

Keywords

  • endocardial cushions
  • endocardium
  • epithelial-mesenchymal transition
  • heparin-binding EGF-like growth factor
  • valve morphogenesis

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

MacGrogan, D., D'Amato, G., Travisano, S., Martinez-Poveda, B., Luxán, G., Del Monte-Nieto, G., ... De La Pompa, J. L. (2016). Sequential Ligand-Dependent Notch Signaling Activation Regulates Valve Primordium Formation and Morphogenesis. Circulation Research, 118(10), 1480-1497. https://doi.org/10.1161/CIRCRESAHA.115.308077

Sequential Ligand-Dependent Notch Signaling Activation Regulates Valve Primordium Formation and Morphogenesis. / MacGrogan, Donal; D'Amato, Gaetano; Travisano, Stanislao; Martinez-Poveda, Beatriz; Luxán, Guillermo; Del Monte-Nieto, Gonzalo; Papoutsi, Tania; Sbroggio, Mauro; Bou, Vanesa; Gomez-Del Arco, Pablo; Gómez, Manuel Jose; Zhou, Bin; Redondo, Juan Miguel; Jiménez-Borreguero, Luis J.; De La Pompa, José Luis.

In: Circulation Research, Vol. 118, No. 10, 13.05.2016, p. 1480-1497.

Research output: Contribution to journalArticle

MacGrogan, D, D'Amato, G, Travisano, S, Martinez-Poveda, B, Luxán, G, Del Monte-Nieto, G, Papoutsi, T, Sbroggio, M, Bou, V, Gomez-Del Arco, P, Gómez, MJ, Zhou, B, Redondo, JM, Jiménez-Borreguero, LJ & De La Pompa, JL 2016, 'Sequential Ligand-Dependent Notch Signaling Activation Regulates Valve Primordium Formation and Morphogenesis', Circulation Research, vol. 118, no. 10, pp. 1480-1497. https://doi.org/10.1161/CIRCRESAHA.115.308077
MacGrogan D, D'Amato G, Travisano S, Martinez-Poveda B, Luxán G, Del Monte-Nieto G et al. Sequential Ligand-Dependent Notch Signaling Activation Regulates Valve Primordium Formation and Morphogenesis. Circulation Research. 2016 May 13;118(10):1480-1497. https://doi.org/10.1161/CIRCRESAHA.115.308077
MacGrogan, Donal ; D'Amato, Gaetano ; Travisano, Stanislao ; Martinez-Poveda, Beatriz ; Luxán, Guillermo ; Del Monte-Nieto, Gonzalo ; Papoutsi, Tania ; Sbroggio, Mauro ; Bou, Vanesa ; Gomez-Del Arco, Pablo ; Gómez, Manuel Jose ; Zhou, Bin ; Redondo, Juan Miguel ; Jiménez-Borreguero, Luis J. ; De La Pompa, José Luis. / Sequential Ligand-Dependent Notch Signaling Activation Regulates Valve Primordium Formation and Morphogenesis. In: Circulation Research. 2016 ; Vol. 118, No. 10. pp. 1480-1497.
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AU - D'Amato, Gaetano

AU - Travisano, Stanislao

AU - Martinez-Poveda, Beatriz

AU - Luxán, Guillermo

AU - Del Monte-Nieto, Gonzalo

AU - Papoutsi, Tania

AU - Sbroggio, Mauro

AU - Bou, Vanesa

AU - Gomez-Del Arco, Pablo

AU - Gómez, Manuel Jose

AU - Zhou, Bin

AU - Redondo, Juan Miguel

AU - Jiménez-Borreguero, Luis J.

AU - De La Pompa, José Luis

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N2 - Rationale: The Notch signaling pathway is crucial for primitive cardiac valve formation by epithelial-mesenchymal transition, and NOTCH1 mutations cause bicuspid aortic valve; however, the temporal requirement for the various Notch ligands and receptors during valve ontogeny is poorly understood. Objective: The aim of this study is to determine the functional specificity of Notch in valve development. Methods and Results: Using cardiac-specific conditional targeted mutant mice, we find that endothelial/endocardial deletion of Mib1-Dll4-Notch1 signaling, possibly favored by Manic-Fringe, is specifically required for cardiac epithelial-mesenchymal transition. Mice lacking endocardial Jag1, Notch1, or RBPJ displayed enlarged valve cusps, bicuspid aortic valve, and septal defects, indicating that endocardial Jag1 to Notch1 signaling is required for post-epithelial-mesenchymal transition valvulogenesis. Valve dysmorphology was associated with increased mesenchyme proliferation, indicating that Jag1-Notch1 signaling restricts mesenchyme cell proliferation non-cell autonomously. Gene profiling revealed upregulated Bmp signaling in Jag1-mutant valves, providing a molecular basis for the hyperproliferative phenotype. Significantly, the negative regulator of mesenchyme proliferation, Hbegf, was markedly reduced in Jag1-mutant valves. Hbegf expression in embryonic endocardial cells could be readily activated through a RBPJ-binding site, identifying Hbegf as an endocardial Notch target. Accordingly, addition of soluble heparin-binding EGF-like growth factor to Jag1-mutant outflow tract explant cultures rescued the hyperproliferative phenotype. Conclusions: During cardiac valve formation, Dll4-Notch1 signaling leads to epithelial-mesenchymal transition and cushion formation. Jag1-Notch1 signaling subsequently restrains Bmp-mediated valve mesenchyme proliferation by sustaining Hbegf-EGF receptor signaling. Our studies identify a mechanism of signaling cross talk during valve morphogenesis involved in the origin of congenital heart defects associated with reduced NOTCH function.

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