PUBLICATION
Exogenous WNT5A and WNT11 proteins rescue CITED2 dysfunction in mouse embryonic stem cells and zebrafish morphants
- Authors
- Santos, J.M.A., Mendes-Silva, L., Afonso, V., Martins, G., Machado, R.S.R., Lopes, J.A., Cancela, L., Futschik, M.E., Sachinidis, A., Gavaia, P., Bragança, J.
- ID
- ZDB-PUB-190807-2
- Date
- 2019
- Source
- Cell Death & Disease 10: 582 (Journal)
- Registered Authors
- Cancela, Leonor, Sachinidis, Agapios
- Keywords
- none
- MeSH Terms
-
- Animals
- Cell Differentiation/genetics
- Cell Line
- Disease Models, Animal
- Female
- Heart Defects, Congenital/metabolism*
- Heart Defects, Congenital/prevention & control
- Male
- Mice
- Mice, Knockout
- Morpholinos/administration & dosage
- Morpholinos/pharmacology
- Mouse Embryonic Stem Cells/metabolism*
- Recombinant Proteins/pharmacology
- Repressor Proteins/antagonists & inhibitors
- Repressor Proteins/genetics
- Repressor Proteins/metabolism*
- Trans-Activators/antagonists & inhibitors
- Trans-Activators/genetics
- Trans-Activators/metabolism*
- Transfection
- Wnt Proteins/pharmacology*
- Wnt-5a Protein/pharmacology*
- Zebrafish/embryology*
- PubMed
- 31378782 Full text @ Cell Death Dis.
Citation
Santos, J.M.A., Mendes-Silva, L., Afonso, V., Martins, G., Machado, R.S.R., Lopes, J.A., Cancela, L., Futschik, M.E., Sachinidis, A., Gavaia, P., Bragança, J. (2019) Exogenous WNT5A and WNT11 proteins rescue CITED2 dysfunction in mouse embryonic stem cells and zebrafish morphants. Cell Death & Disease. 10:582.
Abstract
Mutations and inadequate methylation profiles of CITED2 are associated with human congenital heart disease (CHD). In mouse, Cited2 is necessary for embryogenesis, particularly for heart development, and its depletion in embryonic stem cells (ESC) impairs cardiac differentiation. We have now determined that Cited2 depletion in ESC affects the expression of transcription factors and cardiopoietic genes involved in early mesoderm and cardiac specification. Interestingly, the supplementation of the secretome prepared from ESC overexpressing CITED2, during the onset of differentiation, rescued the cardiogenic defects of Cited2-depleted ESC. In addition, we demonstrate that the proteins WNT5A and WNT11 held the potential for rescue. We also validated the zebrafish as a model to investigate cited2 function during development. Indeed, the microinjection of morpholinos targeting cited2 transcripts caused developmental defects recapitulating those of mice knockout models, including the increased propensity for cardiac defects and severe death rate. Importantly, the co-injection of anti-cited2 morpholinos with either CITED2 or WNT5A and WNT11 recombinant proteins corrected the developmental defects of Cited2-morphants. This study argues that defects caused by the dysfunction of Cited2 at early stages of development, including heart anomalies, may be remediable by supplementation of exogenous molecules, offering the opportunity to develop novel therapeutic strategies aiming to prevent CHD.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping