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Sphingosine 1-phosphate induces differentiation of mesoangioblasts towards smooth muscle. A role for GATA6.

Donati C, Marseglia G, Magi A, Serratì S, Cencetti F, Bernacchioni C, Nannetti G, Benelli M, Brunelli S, Torricelli F, Cossu G, Bruni P - PLoS ONE (2011)

Bottom Line: Quantitative mRNA and protein analysis corroborated the microarray results demonstrating enhanced expression of myogenic marker proteins and regulation of the expression of transcription factor GATA6 and its co-regulator, LMCD1.Finally, the pharmacological inhibition of endogenous S1P formation in response to TGFβ abrogated GATA6 up-regulation, supporting the view that the S1P pathway plays a physiological role in mediating the pro-myogenic effect of TGFβ.This study individuates GATA6 as novel player in the complex transcriptional regulation of mesoangioblast differentiation into SM cells and highlights a role for S1P to favour vascular regeneration.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Scienze Biochimiche, Università di Firenze, Firenze, Italia.

ABSTRACT
Different cells can contribute to repair following vascular injury by differentiating into smooth muscle (SM) cells; however the extracellular signals involved are presently poorly characterized. Mesoangioblasts are progenitor cells capable of differentiating into various mesoderm cell types including SM cells. In this study the biological action exerted by the pleiotropic sphingolipid sphingosine 1-phosphate (S1P) in human mesoangioblasts has been initially investigated by cDNA microarray analysis. Obtained data confirmed the anti-apoptotic action of this sphingolipid and identified for the first time a strong differentiating action toward SM cells. Quantitative mRNA and protein analysis corroborated the microarray results demonstrating enhanced expression of myogenic marker proteins and regulation of the expression of transcription factor GATA6 and its co-regulator, LMCD1. Importantly, GATA6 up-regulation induced by S1P was responsible for the enhanced expression of SM-specific contractile proteins. Moreover, by specific gene silencing experiments GATA6 was critical in the pro-differentiating activity of the cytokine TGFβ. Finally, the pharmacological inhibition of endogenous S1P formation in response to TGFβ abrogated GATA6 up-regulation, supporting the view that the S1P pathway plays a physiological role in mediating the pro-myogenic effect of TGFβ. This study individuates GATA6 as novel player in the complex transcriptional regulation of mesoangioblast differentiation into SM cells and highlights a role for S1P to favour vascular regeneration.

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Related in: MedlinePlus

Dose-dependence of GATA6 expression level induced by S1P (A) and role of the transcription factors GATA6, LMCD1 (B,C) on the expression levels of SM markers induced by S1P in mesoangioblasts.Mutual regulation of the transcription factors GATA6, LMCD1 and TRPS1 (D,E,F). Real time PCR analysis of GATA6 (A) was performed in cells challenged with the indicated concentrations of S1P for 6 h. Real-Time PCR analysis of CNN1 (B), TPM1 (C), GATA6 (D), LMCD1 (E), TRPS1 (F) was performed in cells transfected with scrambled-, GATA6- or LMCD1-siRNA and stimulated with S1P for 24 h.
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pone-0020389-g003: Dose-dependence of GATA6 expression level induced by S1P (A) and role of the transcription factors GATA6, LMCD1 (B,C) on the expression levels of SM markers induced by S1P in mesoangioblasts.Mutual regulation of the transcription factors GATA6, LMCD1 and TRPS1 (D,E,F). Real time PCR analysis of GATA6 (A) was performed in cells challenged with the indicated concentrations of S1P for 6 h. Real-Time PCR analysis of CNN1 (B), TPM1 (C), GATA6 (D), LMCD1 (E), TRPS1 (F) was performed in cells transfected with scrambled-, GATA6- or LMCD1-siRNA and stimulated with S1P for 24 h.

Mentions: Given that GATA6 is recognized as a transcription factor critical for SM differentiation [31], to better characterize the increase of this transcription factor induced by S1P, we next determined if the sphingolipid augmented GATA6 mRNA expression levels in a dose-dependent manner. As shown in Fig. 3A, the sphingolipid was efficacious in upregulating GATA6 mRNA at a concentration of 100 nmol/L up to 1 µmol/L, while was ineffective at a higher concentration, in keeping with the affinity towards S1P exhibited by its receptors.


Sphingosine 1-phosphate induces differentiation of mesoangioblasts towards smooth muscle. A role for GATA6.

Donati C, Marseglia G, Magi A, Serratì S, Cencetti F, Bernacchioni C, Nannetti G, Benelli M, Brunelli S, Torricelli F, Cossu G, Bruni P - PLoS ONE (2011)

Dose-dependence of GATA6 expression level induced by S1P (A) and role of the transcription factors GATA6, LMCD1 (B,C) on the expression levels of SM markers induced by S1P in mesoangioblasts.Mutual regulation of the transcription factors GATA6, LMCD1 and TRPS1 (D,E,F). Real time PCR analysis of GATA6 (A) was performed in cells challenged with the indicated concentrations of S1P for 6 h. Real-Time PCR analysis of CNN1 (B), TPM1 (C), GATA6 (D), LMCD1 (E), TRPS1 (F) was performed in cells transfected with scrambled-, GATA6- or LMCD1-siRNA and stimulated with S1P for 24 h.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3101247&req=5

pone-0020389-g003: Dose-dependence of GATA6 expression level induced by S1P (A) and role of the transcription factors GATA6, LMCD1 (B,C) on the expression levels of SM markers induced by S1P in mesoangioblasts.Mutual regulation of the transcription factors GATA6, LMCD1 and TRPS1 (D,E,F). Real time PCR analysis of GATA6 (A) was performed in cells challenged with the indicated concentrations of S1P for 6 h. Real-Time PCR analysis of CNN1 (B), TPM1 (C), GATA6 (D), LMCD1 (E), TRPS1 (F) was performed in cells transfected with scrambled-, GATA6- or LMCD1-siRNA and stimulated with S1P for 24 h.
Mentions: Given that GATA6 is recognized as a transcription factor critical for SM differentiation [31], to better characterize the increase of this transcription factor induced by S1P, we next determined if the sphingolipid augmented GATA6 mRNA expression levels in a dose-dependent manner. As shown in Fig. 3A, the sphingolipid was efficacious in upregulating GATA6 mRNA at a concentration of 100 nmol/L up to 1 µmol/L, while was ineffective at a higher concentration, in keeping with the affinity towards S1P exhibited by its receptors.

Bottom Line: Quantitative mRNA and protein analysis corroborated the microarray results demonstrating enhanced expression of myogenic marker proteins and regulation of the expression of transcription factor GATA6 and its co-regulator, LMCD1.Finally, the pharmacological inhibition of endogenous S1P formation in response to TGFβ abrogated GATA6 up-regulation, supporting the view that the S1P pathway plays a physiological role in mediating the pro-myogenic effect of TGFβ.This study individuates GATA6 as novel player in the complex transcriptional regulation of mesoangioblast differentiation into SM cells and highlights a role for S1P to favour vascular regeneration.

View Article: PubMed Central - PubMed

Affiliation: Dipartimento di Scienze Biochimiche, Università di Firenze, Firenze, Italia.

ABSTRACT
Different cells can contribute to repair following vascular injury by differentiating into smooth muscle (SM) cells; however the extracellular signals involved are presently poorly characterized. Mesoangioblasts are progenitor cells capable of differentiating into various mesoderm cell types including SM cells. In this study the biological action exerted by the pleiotropic sphingolipid sphingosine 1-phosphate (S1P) in human mesoangioblasts has been initially investigated by cDNA microarray analysis. Obtained data confirmed the anti-apoptotic action of this sphingolipid and identified for the first time a strong differentiating action toward SM cells. Quantitative mRNA and protein analysis corroborated the microarray results demonstrating enhanced expression of myogenic marker proteins and regulation of the expression of transcription factor GATA6 and its co-regulator, LMCD1. Importantly, GATA6 up-regulation induced by S1P was responsible for the enhanced expression of SM-specific contractile proteins. Moreover, by specific gene silencing experiments GATA6 was critical in the pro-differentiating activity of the cytokine TGFβ. Finally, the pharmacological inhibition of endogenous S1P formation in response to TGFβ abrogated GATA6 up-regulation, supporting the view that the S1P pathway plays a physiological role in mediating the pro-myogenic effect of TGFβ. This study individuates GATA6 as novel player in the complex transcriptional regulation of mesoangioblast differentiation into SM cells and highlights a role for S1P to favour vascular regeneration.

Show MeSH
Related in: MedlinePlus