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Tumor-derived mural-like cells coordinate with endothelial cells: role of YKL-40 in mural cell-mediated angiogenesis.

Francescone R, Ngernyuang N, Yan W, Bentley B, Shao R - Oncogene (2013)

Bottom Line: YKL-40 expressed by GSDCs was associated with increased interaction of neural cadherin/β-catenin/smooth muscle alpha actin; thus, mediating cell-cell adhesion and permeability.In cell co-culture systems, YKL-40 enhanced both GSDC and HMVEC contacts, restricted vascular leakage, and stabilized vascular networks.Collectively, the data inform new mechanistic insights into the cooperation of mural cells with endothelial cells induced by YKL-40 during tumor angiogenesis, and also enhance our understanding of YKL-40 in both mural and endothelial cell biology.

View Article: PubMed Central - PubMed

Affiliation: Molecular and Cellular Biology Program, Morrill Science Center, University of Massachusetts, Amherst, MA, USA.

ABSTRACT
Tumor neo-vasculature is characterized by spatial coordination of endothelial cells with mural cells, which delivers oxygen and nutrients. Here, we explored a key role of the secreted glycoprotein YKL-40, a mesenchymal marker, in the interaction between endothelial cells and mesenchymal mural-like cells for tumor angiogenesis. Xenotransplantation of tumor-derived mural-like cells (GSDCs) expressing YKL-40 in mice developed extensive and stable blood vessels covered with more GSDCs than those in YKL-40 gene knockdown tumors. YKL-40 expressed by GSDCs was associated with increased interaction of neural cadherin/β-catenin/smooth muscle alpha actin; thus, mediating cell-cell adhesion and permeability. YKL-40 also induced the interaction of vascular endothelial cadherin/β-catenin/actin in endothelial cells (HMVECs). In cell co-culture systems, YKL-40 enhanced both GSDC and HMVEC contacts, restricted vascular leakage, and stabilized vascular networks. Collectively, the data inform new mechanistic insights into the cooperation of mural cells with endothelial cells induced by YKL-40 during tumor angiogenesis, and also enhance our understanding of YKL-40 in both mural and endothelial cell biology.

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Co-culture of HMVECs and YKL-40-expressing GSDCs displays interaction between cadherin and cateninA. (Top) HMVECs cultured with control or YKL-40 shRNA GSDCs were stained for VE-cad (red) and β-cate (green). White arrowheads mark regions of co-localization of VE-cad and β-cate. (Middle) VE-cad (red) staining was to show HMVECs distinct from VE-cad-negative GSDCs. White asterisks indicate HMVECs. DAPI (blue) was used as a nuclear stain for all images. (Bottom) Quantification of the percentage of HMVECs that have co-expression (yellow) of both VE-cad and β-cate in a total of HMVECs. Bars: 10 μm. N=3, *P≤0.05 compared to control. B. (Top) HMVECs and either control or YKL-40 shRNA GSDCs were cultured together and tested for overlapping of N-cad (green) and β-cate (red). DAPI was pre-stained for the nuclei of GSDCs only, but not for HMVECs to discriminate different cell types. (Bottom, Left) Representative images of the different types of cell to cell contacts: GSDC-GSDC (white arrowhead), HMVEC-HMVEC (black arrowhead), and HMVEC-GSDC (white arrow). (Bottom, right) Quantification of the percentage of cells containing N-cad/β-cate overlapping contacts in a total of individual cell types, displayed by cell contact types. Bars: 10 μm. N=3, *P≤0.05 compared to control.
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Figure 5: Co-culture of HMVECs and YKL-40-expressing GSDCs displays interaction between cadherin and cateninA. (Top) HMVECs cultured with control or YKL-40 shRNA GSDCs were stained for VE-cad (red) and β-cate (green). White arrowheads mark regions of co-localization of VE-cad and β-cate. (Middle) VE-cad (red) staining was to show HMVECs distinct from VE-cad-negative GSDCs. White asterisks indicate HMVECs. DAPI (blue) was used as a nuclear stain for all images. (Bottom) Quantification of the percentage of HMVECs that have co-expression (yellow) of both VE-cad and β-cate in a total of HMVECs. Bars: 10 μm. N=3, *P≤0.05 compared to control. B. (Top) HMVECs and either control or YKL-40 shRNA GSDCs were cultured together and tested for overlapping of N-cad (green) and β-cate (red). DAPI was pre-stained for the nuclei of GSDCs only, but not for HMVECs to discriminate different cell types. (Bottom, Left) Representative images of the different types of cell to cell contacts: GSDC-GSDC (white arrowhead), HMVEC-HMVEC (black arrowhead), and HMVEC-GSDC (white arrow). (Bottom, right) Quantification of the percentage of cells containing N-cad/β-cate overlapping contacts in a total of individual cell types, displayed by cell contact types. Bars: 10 μm. N=3, *P≤0.05 compared to control.

Mentions: To monitor the interaction of cadherin and catenin between endothelial cells and mural cells that may recapitulate their functional relationship in vessels in vivo, we undertook an immunocytochemical approach probing the co-localization of either VE-cadherin/β-catenin or N-cadherin/β-catenin in a cell co-culture system with GSDCs and HMVECs. Intense co-staining of VE-cadherin/β-catenin was found in HMVECs mixed with control GSDCs, but this co-localization was decreased by 58% in HMVECs when co-cultured with YKL-40 shRNA GSDCs (Fig. 5A), in which HMVECs were distinguished from GSDCs by positive staining of VE-cadherin as indicated with white asterisks (Fig. 5A). This is indicative that YKL-40 expressing cells are crucial to VE-cadherin/β-catenin interaction in HMVECs.


Tumor-derived mural-like cells coordinate with endothelial cells: role of YKL-40 in mural cell-mediated angiogenesis.

Francescone R, Ngernyuang N, Yan W, Bentley B, Shao R - Oncogene (2013)

Co-culture of HMVECs and YKL-40-expressing GSDCs displays interaction between cadherin and cateninA. (Top) HMVECs cultured with control or YKL-40 shRNA GSDCs were stained for VE-cad (red) and β-cate (green). White arrowheads mark regions of co-localization of VE-cad and β-cate. (Middle) VE-cad (red) staining was to show HMVECs distinct from VE-cad-negative GSDCs. White asterisks indicate HMVECs. DAPI (blue) was used as a nuclear stain for all images. (Bottom) Quantification of the percentage of HMVECs that have co-expression (yellow) of both VE-cad and β-cate in a total of HMVECs. Bars: 10 μm. N=3, *P≤0.05 compared to control. B. (Top) HMVECs and either control or YKL-40 shRNA GSDCs were cultured together and tested for overlapping of N-cad (green) and β-cate (red). DAPI was pre-stained for the nuclei of GSDCs only, but not for HMVECs to discriminate different cell types. (Bottom, Left) Representative images of the different types of cell to cell contacts: GSDC-GSDC (white arrowhead), HMVEC-HMVEC (black arrowhead), and HMVEC-GSDC (white arrow). (Bottom, right) Quantification of the percentage of cells containing N-cad/β-cate overlapping contacts in a total of individual cell types, displayed by cell contact types. Bars: 10 μm. N=3, *P≤0.05 compared to control.
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Figure 5: Co-culture of HMVECs and YKL-40-expressing GSDCs displays interaction between cadherin and cateninA. (Top) HMVECs cultured with control or YKL-40 shRNA GSDCs were stained for VE-cad (red) and β-cate (green). White arrowheads mark regions of co-localization of VE-cad and β-cate. (Middle) VE-cad (red) staining was to show HMVECs distinct from VE-cad-negative GSDCs. White asterisks indicate HMVECs. DAPI (blue) was used as a nuclear stain for all images. (Bottom) Quantification of the percentage of HMVECs that have co-expression (yellow) of both VE-cad and β-cate in a total of HMVECs. Bars: 10 μm. N=3, *P≤0.05 compared to control. B. (Top) HMVECs and either control or YKL-40 shRNA GSDCs were cultured together and tested for overlapping of N-cad (green) and β-cate (red). DAPI was pre-stained for the nuclei of GSDCs only, but not for HMVECs to discriminate different cell types. (Bottom, Left) Representative images of the different types of cell to cell contacts: GSDC-GSDC (white arrowhead), HMVEC-HMVEC (black arrowhead), and HMVEC-GSDC (white arrow). (Bottom, right) Quantification of the percentage of cells containing N-cad/β-cate overlapping contacts in a total of individual cell types, displayed by cell contact types. Bars: 10 μm. N=3, *P≤0.05 compared to control.
Mentions: To monitor the interaction of cadherin and catenin between endothelial cells and mural cells that may recapitulate their functional relationship in vessels in vivo, we undertook an immunocytochemical approach probing the co-localization of either VE-cadherin/β-catenin or N-cadherin/β-catenin in a cell co-culture system with GSDCs and HMVECs. Intense co-staining of VE-cadherin/β-catenin was found in HMVECs mixed with control GSDCs, but this co-localization was decreased by 58% in HMVECs when co-cultured with YKL-40 shRNA GSDCs (Fig. 5A), in which HMVECs were distinguished from GSDCs by positive staining of VE-cadherin as indicated with white asterisks (Fig. 5A). This is indicative that YKL-40 expressing cells are crucial to VE-cadherin/β-catenin interaction in HMVECs.

Bottom Line: YKL-40 expressed by GSDCs was associated with increased interaction of neural cadherin/β-catenin/smooth muscle alpha actin; thus, mediating cell-cell adhesion and permeability.In cell co-culture systems, YKL-40 enhanced both GSDC and HMVEC contacts, restricted vascular leakage, and stabilized vascular networks.Collectively, the data inform new mechanistic insights into the cooperation of mural cells with endothelial cells induced by YKL-40 during tumor angiogenesis, and also enhance our understanding of YKL-40 in both mural and endothelial cell biology.

View Article: PubMed Central - PubMed

Affiliation: Molecular and Cellular Biology Program, Morrill Science Center, University of Massachusetts, Amherst, MA, USA.

ABSTRACT
Tumor neo-vasculature is characterized by spatial coordination of endothelial cells with mural cells, which delivers oxygen and nutrients. Here, we explored a key role of the secreted glycoprotein YKL-40, a mesenchymal marker, in the interaction between endothelial cells and mesenchymal mural-like cells for tumor angiogenesis. Xenotransplantation of tumor-derived mural-like cells (GSDCs) expressing YKL-40 in mice developed extensive and stable blood vessels covered with more GSDCs than those in YKL-40 gene knockdown tumors. YKL-40 expressed by GSDCs was associated with increased interaction of neural cadherin/β-catenin/smooth muscle alpha actin; thus, mediating cell-cell adhesion and permeability. YKL-40 also induced the interaction of vascular endothelial cadherin/β-catenin/actin in endothelial cells (HMVECs). In cell co-culture systems, YKL-40 enhanced both GSDC and HMVEC contacts, restricted vascular leakage, and stabilized vascular networks. Collectively, the data inform new mechanistic insights into the cooperation of mural cells with endothelial cells induced by YKL-40 during tumor angiogenesis, and also enhance our understanding of YKL-40 in both mural and endothelial cell biology.

Show MeSH
Related in: MedlinePlus