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Mesenchymal-endothelial transition contributes to cardiac neovascularization.

Ubil E, Duan J, Pillai IC, Rosa-Garrido M, Wu Y, Bargiacchi F, Lu Y, Stanbouly S, Huang J, Rojas M, Vondriska TM, Stefani E, Deb A - Nature (2014)

Bottom Line: We show that the transcription factor p53 regulates such a switch in cardiac fibroblast fate.Loss of p53 in cardiac fibroblasts severely decreases the formation of fibroblast-derived endothelial cells, reduces post-infarct vascular density and worsens cardiac function.These observations demonstrate that mesenchymal-to-endothelial transition contributes to neovascularization of the injured heart and represents a potential therapeutic target for enhancing cardiac repair.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology &Physiology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599, USA.

ABSTRACT
Endothelial cells contribute to a subset of cardiac fibroblasts by undergoing endothelial-to-mesenchymal transition, but whether cardiac fibroblasts can adopt an endothelial cell fate and directly contribute to neovascularization after cardiac injury is not known. Here, using genetic fate map techniques, we demonstrate that cardiac fibroblasts rapidly adopt an endothelial-cell-like phenotype after acute ischaemic cardiac injury. Fibroblast-derived endothelial cells exhibit anatomical and functional characteristics of native endothelial cells. We show that the transcription factor p53 regulates such a switch in cardiac fibroblast fate. Loss of p53 in cardiac fibroblasts severely decreases the formation of fibroblast-derived endothelial cells, reduces post-infarct vascular density and worsens cardiac function. Conversely, stimulation of the p53 pathway in cardiac fibroblasts augments mesenchymal-to-endothelial transition, enhances vascularity and improves cardiac function. These observations demonstrate that mesenchymal-to-endothelial transition contributes to neovascularization of the injured heart and represents a potential therapeutic target for enhancing cardiac repair.

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Effect of serum starvation on p53 levels and effects of Pifithrin-α and RITA on tube formation of serum fed cardiac fibroblasts(a) Western blot for p53 in cardiac fibroblasts subjected to serum starvation for 24, 48 and 72 hours (representative sample from n=3) (b) densitometric quantitation of Western blot (mean±S.E.M., *p<0.05 compared to cells in 10% serum). (c) Effect on tube formation after adding Pifithrin-α or RITA to cardiac fibroblasts grown in 10% serum Scale bar: 250μm (d) quantitation of tube formation (mean±S.E.M., *p<0.05, n=3)
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Figure 11: Effect of serum starvation on p53 levels and effects of Pifithrin-α and RITA on tube formation of serum fed cardiac fibroblasts(a) Western blot for p53 in cardiac fibroblasts subjected to serum starvation for 24, 48 and 72 hours (representative sample from n=3) (b) densitometric quantitation of Western blot (mean±S.E.M., *p<0.05 compared to cells in 10% serum). (c) Effect on tube formation after adding Pifithrin-α or RITA to cardiac fibroblasts grown in 10% serum Scale bar: 250μm (d) quantitation of tube formation (mean±S.E.M., *p<0.05, n=3)

Mentions: To determine whether p53 plays a regulatory role in MEndoT, we first established an ex vivo model. Cells subjected to types of cellular stress ex vivo, such as serum deprivation, upregulate p53 levels27,28. We subjected cardiac fibroblasts to serum starvation and observed increased p53 levels by Western Blotting (Extended Data Fig. 7a,b). We next seeded labeled cardiac fibroblasts (99% purity by flow cytometry), on matrigel (a mixture of basement membrane proteins that facilitates capillary tube formation) and subjected them to serum starvation. In contrast to control fibroblasts (Fig. 2e,g) serum starved fibroblasts formed capillary tube like structures, expressed VECAD (Fig. 2f) and took up AcLDL (Fig. 2h), consistent with adoption of an endothelial cell like phenotype.


Mesenchymal-endothelial transition contributes to cardiac neovascularization.

Ubil E, Duan J, Pillai IC, Rosa-Garrido M, Wu Y, Bargiacchi F, Lu Y, Stanbouly S, Huang J, Rojas M, Vondriska TM, Stefani E, Deb A - Nature (2014)

Effect of serum starvation on p53 levels and effects of Pifithrin-α and RITA on tube formation of serum fed cardiac fibroblasts(a) Western blot for p53 in cardiac fibroblasts subjected to serum starvation for 24, 48 and 72 hours (representative sample from n=3) (b) densitometric quantitation of Western blot (mean±S.E.M., *p<0.05 compared to cells in 10% serum). (c) Effect on tube formation after adding Pifithrin-α or RITA to cardiac fibroblasts grown in 10% serum Scale bar: 250μm (d) quantitation of tube formation (mean±S.E.M., *p<0.05, n=3)
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4214889&req=5

Figure 11: Effect of serum starvation on p53 levels and effects of Pifithrin-α and RITA on tube formation of serum fed cardiac fibroblasts(a) Western blot for p53 in cardiac fibroblasts subjected to serum starvation for 24, 48 and 72 hours (representative sample from n=3) (b) densitometric quantitation of Western blot (mean±S.E.M., *p<0.05 compared to cells in 10% serum). (c) Effect on tube formation after adding Pifithrin-α or RITA to cardiac fibroblasts grown in 10% serum Scale bar: 250μm (d) quantitation of tube formation (mean±S.E.M., *p<0.05, n=3)
Mentions: To determine whether p53 plays a regulatory role in MEndoT, we first established an ex vivo model. Cells subjected to types of cellular stress ex vivo, such as serum deprivation, upregulate p53 levels27,28. We subjected cardiac fibroblasts to serum starvation and observed increased p53 levels by Western Blotting (Extended Data Fig. 7a,b). We next seeded labeled cardiac fibroblasts (99% purity by flow cytometry), on matrigel (a mixture of basement membrane proteins that facilitates capillary tube formation) and subjected them to serum starvation. In contrast to control fibroblasts (Fig. 2e,g) serum starved fibroblasts formed capillary tube like structures, expressed VECAD (Fig. 2f) and took up AcLDL (Fig. 2h), consistent with adoption of an endothelial cell like phenotype.

Bottom Line: We show that the transcription factor p53 regulates such a switch in cardiac fibroblast fate.Loss of p53 in cardiac fibroblasts severely decreases the formation of fibroblast-derived endothelial cells, reduces post-infarct vascular density and worsens cardiac function.These observations demonstrate that mesenchymal-to-endothelial transition contributes to neovascularization of the injured heart and represents a potential therapeutic target for enhancing cardiac repair.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology &Physiology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599, USA.

ABSTRACT
Endothelial cells contribute to a subset of cardiac fibroblasts by undergoing endothelial-to-mesenchymal transition, but whether cardiac fibroblasts can adopt an endothelial cell fate and directly contribute to neovascularization after cardiac injury is not known. Here, using genetic fate map techniques, we demonstrate that cardiac fibroblasts rapidly adopt an endothelial-cell-like phenotype after acute ischaemic cardiac injury. Fibroblast-derived endothelial cells exhibit anatomical and functional characteristics of native endothelial cells. We show that the transcription factor p53 regulates such a switch in cardiac fibroblast fate. Loss of p53 in cardiac fibroblasts severely decreases the formation of fibroblast-derived endothelial cells, reduces post-infarct vascular density and worsens cardiac function. Conversely, stimulation of the p53 pathway in cardiac fibroblasts augments mesenchymal-to-endothelial transition, enhances vascularity and improves cardiac function. These observations demonstrate that mesenchymal-to-endothelial transition contributes to neovascularization of the injured heart and represents a potential therapeutic target for enhancing cardiac repair.

Show MeSH
Related in: MedlinePlus