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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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Flow cytometry for endothelial progenitor markers on non-myocyte cells harvested from uninjured hearts of Col1a2CreERT:R26RtdTomato mice and bone marrow cells isolated from the same animal(a,b) Hearts of mice were digested, myocytes discarded and the entire non-myocyte population without any further selection was subjected to flow cytometry. Expression of tdTomato and (a) CD45 and Flk-1 (APC fluorophore) and (b) CD133, CD34 and combined expression of CD34 and Flk-1 in non-myocyte population. (c–f) Bone marrow cells were isolated from Col1a2CreERT:R26RtdTomato mice and without further culture subjected to flow cytometry. Expression of tdTomato and (c) CD45 (d) Flk-1 (e) CD133 (f) CD34 and combined expression of Flk1 and CD34 in bone marrow cells (g) Expression of tdTomato in bone marrow derived mesenchymal stem cell colonies (black arrowhead points to rare tdTomato positive cell, white arrowheads point to mesenchymal stem cells). Scale bar: 50μm.
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Figure 9: Flow cytometry for endothelial progenitor markers on non-myocyte cells harvested from uninjured hearts of Col1a2CreERT:R26RtdTomato mice and bone marrow cells isolated from the same animal(a,b) Hearts of mice were digested, myocytes discarded and the entire non-myocyte population without any further selection was subjected to flow cytometry. Expression of tdTomato and (a) CD45 and Flk-1 (APC fluorophore) and (b) CD133, CD34 and combined expression of CD34 and Flk-1 in non-myocyte population. (c–f) Bone marrow cells were isolated from Col1a2CreERT:R26RtdTomato mice and without further culture subjected to flow cytometry. Expression of tdTomato and (c) CD45 (d) Flk-1 (e) CD133 (f) CD34 and combined expression of Flk1 and CD34 in bone marrow cells (g) Expression of tdTomato in bone marrow derived mesenchymal stem cell colonies (black arrowhead points to rare tdTomato positive cell, white arrowheads point to mesenchymal stem cells). Scale bar: 50μm.

Mentions: Next we investigated whether Col1a2 expressing endothelial progenitors in the heart or bone marrow could have been labeled during administration of tamoxifen and subsequently generated tdTomato+ endothelial cells. We analyzed the heart and bone marrow but observed less than 0.5% of tdTomato cells in the heart (Extended Data Fig. 5a,b) or bone marrow (Extended Data Fig. 5c–f) to express endothelial progenitor markers (CD133 CD34 and Flk1)23. Bone marrow derived mesenchymal stem cells did not exhibit tdTomato fluorescence with the exception of rare cells (Extended Data Fig. 5g). Collectively, these observations suggest that substantial labeling of endothelial progenitors did not occur making them an unlikely source of tdTomato+ endothelial cells.


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)

Flow cytometry for endothelial progenitor markers on non-myocyte cells harvested from uninjured hearts of Col1a2CreERT:R26RtdTomato mice and bone marrow cells isolated from the same animal(a,b) Hearts of mice were digested, myocytes discarded and the entire non-myocyte population without any further selection was subjected to flow cytometry. Expression of tdTomato and (a) CD45 and Flk-1 (APC fluorophore) and (b) CD133, CD34 and combined expression of CD34 and Flk-1 in non-myocyte population. (c–f) Bone marrow cells were isolated from Col1a2CreERT:R26RtdTomato mice and without further culture subjected to flow cytometry. Expression of tdTomato and (c) CD45 (d) Flk-1 (e) CD133 (f) CD34 and combined expression of Flk1 and CD34 in bone marrow cells (g) Expression of tdTomato in bone marrow derived mesenchymal stem cell colonies (black arrowhead points to rare tdTomato positive cell, white arrowheads point to mesenchymal stem cells). Scale bar: 50μm.
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Related In: Results  -  Collection

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

Figure 9: Flow cytometry for endothelial progenitor markers on non-myocyte cells harvested from uninjured hearts of Col1a2CreERT:R26RtdTomato mice and bone marrow cells isolated from the same animal(a,b) Hearts of mice were digested, myocytes discarded and the entire non-myocyte population without any further selection was subjected to flow cytometry. Expression of tdTomato and (a) CD45 and Flk-1 (APC fluorophore) and (b) CD133, CD34 and combined expression of CD34 and Flk-1 in non-myocyte population. (c–f) Bone marrow cells were isolated from Col1a2CreERT:R26RtdTomato mice and without further culture subjected to flow cytometry. Expression of tdTomato and (c) CD45 (d) Flk-1 (e) CD133 (f) CD34 and combined expression of Flk1 and CD34 in bone marrow cells (g) Expression of tdTomato in bone marrow derived mesenchymal stem cell colonies (black arrowhead points to rare tdTomato positive cell, white arrowheads point to mesenchymal stem cells). Scale bar: 50μm.
Mentions: Next we investigated whether Col1a2 expressing endothelial progenitors in the heart or bone marrow could have been labeled during administration of tamoxifen and subsequently generated tdTomato+ endothelial cells. We analyzed the heart and bone marrow but observed less than 0.5% of tdTomato cells in the heart (Extended Data Fig. 5a,b) or bone marrow (Extended Data Fig. 5c–f) to express endothelial progenitor markers (CD133 CD34 and Flk1)23. Bone marrow derived mesenchymal stem cells did not exhibit tdTomato fluorescence with the exception of rare cells (Extended Data Fig. 5g). Collectively, these observations suggest that substantial labeling of endothelial progenitors did not occur making them an unlikely source of tdTomato+ endothelial cells.

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