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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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YKL-40 enhances interaction of VE-cadherin and β-catenin, but VEGF attenuates the interaction in HMVECsA. (Top) HMEVC lysates treated 24 hr with serum-free medium (SFM) with or without 10 ng/mL VEGF, or 200 ng/mL recombinant YKL-40 were immunoprecipated with an anti-VE-cad Ab and probed for β-cate by western blotting. IgG was used as IP control. (Bottom) Quantification of the western blots above, normalized to IgG levels. *P<0.05 compared with SFM. n=3. B. Representative images of VE-cad (red) and β-cate (green) double stained HMVECs in the presence of either SFM, 10 ng/mL VEGF, or 200 ng/mL for 24 hr. White arrowheads highlight the areas positively co-stained for VE-cad and β-cate (yellow). C. Quantification of the VE-cad/β-cate overlap in the images in part C. A bar: 20 μm. N=3, *P≤0.05 compared to control.
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Figure 4: YKL-40 enhances interaction of VE-cadherin and β-catenin, but VEGF attenuates the interaction in HMVECsA. (Top) HMEVC lysates treated 24 hr with serum-free medium (SFM) with or without 10 ng/mL VEGF, or 200 ng/mL recombinant YKL-40 were immunoprecipated with an anti-VE-cad Ab and probed for β-cate by western blotting. IgG was used as IP control. (Bottom) Quantification of the western blots above, normalized to IgG levels. *P<0.05 compared with SFM. n=3. B. Representative images of VE-cad (red) and β-cate (green) double stained HMVECs in the presence of either SFM, 10 ng/mL VEGF, or 200 ng/mL for 24 hr. White arrowheads highlight the areas positively co-stained for VE-cad and β-cate (yellow). C. Quantification of the VE-cad/β-cate overlap in the images in part C. A bar: 20 μm. N=3, *P≤0.05 compared to control.

Mentions: In order to investigate the roles that YKL-40 and VEGF play individually in the association between VE-cadherin and β-catenin in HMVECs, we treated the cells with recombinant protein of VEGF or YKL-40. Treatment of HMVECs with YKL-40 or VEGF did not alter protein expression of VE-cadherin, N-cadherin, or β-catenin (data not shown). However, YKL-40 induced the interaction between VE-cadherin and β-catenin; in contrast, VEGF inhibited their association (Fig. 4A). The immunocytochemical analysis validated this result that YKL-40 significantly increased co-localization of VE-cadherin and β-catenin by 60% relative to the control, while VEGF treatment decreased their co-localization by 43% (Fig. 4B & 4C). In a trial using different concentrations of these proteins, VEGF (10 ng/ml) and YKL-40 (200 ng/ml) at the pathologic levels were found to have more effects on these adhesion molecule interactions than other concentrations (data not shown). The results indicate that YKL-40, in contrast to VEGF, induces the interaction of VE-cadherin and β-catenin in HMVECs, further supporting the earlier findings using GSDC-conditioned media expressing YKL-40.


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)

YKL-40 enhances interaction of VE-cadherin and β-catenin, but VEGF attenuates the interaction in HMVECsA. (Top) HMEVC lysates treated 24 hr with serum-free medium (SFM) with or without 10 ng/mL VEGF, or 200 ng/mL recombinant YKL-40 were immunoprecipated with an anti-VE-cad Ab and probed for β-cate by western blotting. IgG was used as IP control. (Bottom) Quantification of the western blots above, normalized to IgG levels. *P<0.05 compared with SFM. n=3. B. Representative images of VE-cad (red) and β-cate (green) double stained HMVECs in the presence of either SFM, 10 ng/mL VEGF, or 200 ng/mL for 24 hr. White arrowheads highlight the areas positively co-stained for VE-cad and β-cate (yellow). C. Quantification of the VE-cad/β-cate overlap in the images in part C. A bar: 20 μm. N=3, *P≤0.05 compared to control.
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Figure 4: YKL-40 enhances interaction of VE-cadherin and β-catenin, but VEGF attenuates the interaction in HMVECsA. (Top) HMEVC lysates treated 24 hr with serum-free medium (SFM) with or without 10 ng/mL VEGF, or 200 ng/mL recombinant YKL-40 were immunoprecipated with an anti-VE-cad Ab and probed for β-cate by western blotting. IgG was used as IP control. (Bottom) Quantification of the western blots above, normalized to IgG levels. *P<0.05 compared with SFM. n=3. B. Representative images of VE-cad (red) and β-cate (green) double stained HMVECs in the presence of either SFM, 10 ng/mL VEGF, or 200 ng/mL for 24 hr. White arrowheads highlight the areas positively co-stained for VE-cad and β-cate (yellow). C. Quantification of the VE-cad/β-cate overlap in the images in part C. A bar: 20 μm. N=3, *P≤0.05 compared to control.
Mentions: In order to investigate the roles that YKL-40 and VEGF play individually in the association between VE-cadherin and β-catenin in HMVECs, we treated the cells with recombinant protein of VEGF or YKL-40. Treatment of HMVECs with YKL-40 or VEGF did not alter protein expression of VE-cadherin, N-cadherin, or β-catenin (data not shown). However, YKL-40 induced the interaction between VE-cadherin and β-catenin; in contrast, VEGF inhibited their association (Fig. 4A). The immunocytochemical analysis validated this result that YKL-40 significantly increased co-localization of VE-cadherin and β-catenin by 60% relative to the control, while VEGF treatment decreased their co-localization by 43% (Fig. 4B & 4C). In a trial using different concentrations of these proteins, VEGF (10 ng/ml) and YKL-40 (200 ng/ml) at the pathologic levels were found to have more effects on these adhesion molecule interactions than other concentrations (data not shown). The results indicate that YKL-40, in contrast to VEGF, induces the interaction of VE-cadherin and β-catenin in HMVECs, further supporting the earlier findings using GSDC-conditioned media expressing YKL-40.

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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Related in: MedlinePlus