The requirement for fibroblasts in angiogenesis: fibroblast-derived matrix proteins are essential for endothelial cell lumen formation
| Autor: | Martin N. Nakatsu, Andrew C. Newman, Christopher C.W. Hughes, Wayne Chou, Paul D. Gershon |
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| Rok vydání: | 2011 |
| Předmět: |
Angiogenesis
Gene Expression Pregnancy Proteins Mass Spectrometry Extracellular matrix 0302 clinical medicine Transforming Growth Factor beta Cell Interactions Angiogenic Proteins RNA Small Interfering Chromatography Extracellular Matrix Proteins 0303 health sciences Neovascularization Pathologic Intracellular Signaling Peptides and Proteins Articles Extracellular Matrix Cell biology Endothelial stem cell medicine.anatomical_structure Gene Knockdown Techniques 030220 oncology & carcinogenesis Hepatocyte growth factor Collagen Rheology Cell Division Signal Transduction medicine.drug Angiogenin Neovascularization Physiologic macromolecular substances Biology 03 medical and health sciences Cell Line Tumor Human Umbilical Vein Endothelial Cells medicine Humans Fibroblast Molecular Biology Glycoproteins 030304 developmental biology Tumor microenvironment Cell Biology Transforming growth factor beta Fibroblasts Molecular biology Glycodelin Culture Media Conditioned biology.protein Carrier Proteins |
| Zdroj: | Molecular Biology of the Cell Newman, AC; Nakatsu, MN; Chou, W; Gershon, PD; & Hughes, CCW. (2011). The requirement for fibroblasts in angiogenesis: Fibroblast-derived matrix proteins are essential for endothelial cell lumen formation. Molecular Biology of the Cell, 22(20), 3791-3800. doi: 10.1091/mbc.E11-05-0393. UC Irvine: Retrieved from: http://www.escholarship.org/uc/item/70k9g8pz |
| ISSN: | 1939-4586 1059-1524 |
| DOI: | 10.1091/mbc.e11-05-0393 |
| Popis: | The combination of a candidate gene approach, column chromatography, and mass spectrometry identifies several fibroblast-derived proteins essential for endothelial cell sprouting and lumen formation. Furthermore, proteins responsible for EC lumen formation increase matrix stiffness, which correlates with EC lumenogenesis. A role for fibroblasts in physiological and pathological angiogenesis is now well recognized; however, the precise mechanisms underlying their action have not been determined. Using an in vitro angiogenesis model in combination with a candidate gene approach, column chromatography, and mass spectrometry, we identify two classes of fibroblast-derived factors—one that supports vessel sprouting but not lumen formation, and one that promotes lumen formation. In the absence of fibroblasts a combination of angiopoietin-1, angiogenin, hepatocyte growth factor, transforming growth factor-α, and tumor necrosis factor drives robust endothelial cell (EC) sprouting; however, lumens fail to form. Subsequent addition of fibroblast-conditioned medium restores lumenogenesis. Using small interfering RNA–mediated knockdown, we show that five genes expressed in fibroblasts—collagen I, procollagen C endopeptidase enhancer 1, secreted protein acidic and rich in cysteine, transforming growth factor-β–induced protein ig-h3, and insulin growth factor–binding protein 7—are necessary for lumen formation. Moreover, lumen formation can be rescued by addition of purified protein to knockdown cultures. Finally, using rheology, we demonstrate that the presence of these matricellular proteins results in significantly stiffer gels, which correlates with enhanced lumen formation. These findings highlight the critical role that fibroblast-derived extracellular matrix components play in EC lumen formation and provide potential insight into the role of fibroblasts in the tumor microenvironment. |
| Databáze: | OpenAIRE |
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