title: A heterogeneous in vitro three dimensional model of tumour-stroma interactions regulating sprouting angiogenesis.
abstract: Angiogenesis, the formation of new blood vessels, is an essential process for tumour progression and is an area of significant therapeutic interest. Different in vitro systems and more complex in vivo systems have been described for the study of tumour angiogenesis. However, there are few human 3D in vitro systems described to date which mimic the cellular heterogeneity and complexity of angiogenesis within the tumour microenvironment. In this study we describe the Minitumour model--a 3 dimensional human spheroid-based system consisting of endothelial cells and fibroblasts in co-culture with the breast cancer cell line MDA-MB-231, for the study of tumour angiogenesis in vitro. After implantation in collagen-I gels, Minitumour spheroids form quantifiable endothelial capillary-like structures. The endothelial cell pre-capillary sprouts are supported by the fibroblasts, which act as mural cells, and their growth is increased by the presence of cancer cells. Characterisation of the Minitumour model using small molecule inhibitors and inhibitory antibodies show that endothelial sprout formation is dependent on growth factors and cytokines known to be important for tumour angiogenesis. The model also shows a response to anti-angiogenic agents similar to previously described in vivo data. We demonstrate that independent manipulation of the different cell types is possible, using common molecular techniques, before incorporation into the model. This aspect of Minitumour spheroid analysis makes this model ideal for high content studies of gene function in individual cell types, allowing for the dissection of their roles in cell-cell interactions. Finally, using this technique, we were able to show the requirement of the metalloproteinase MT1-MMP in endothelial cells and fibroblasts, but not cancer cells, for sprouting angiogenesis.
authors.affiliation: University of Cambridge, Department of Oncology, Cancer Research UK Cambridge Research Institute, Li Ka Shing Centre, Cambridge, United Kingdom.
authors.forename: Pedro
authors.initials: P
authors.lastname: Correa de Sampaio
authors.forename: David
authors.initials: D
authors.lastname: Auslaender
authors.forename: Davia
authors.initials: D
authors.lastname: Krubasik
authors.forename: Antonio Virgilio
authors.initials: AV
authors.lastname: Failla
authors.forename: Jeremy N
authors.initials: JN
authors.lastname: Skepper
authors.forename: Gillian
authors.initials: G
authors.lastname: Murphy
authors.forename: William R
authors.initials: WR
authors.lastname: English
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D002450:Cell Communication / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D020868:Gene Silencing / Q000187:drug effects; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D008163:Luminescent Measurements; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism; D036641:Microscopy, Fluorescence, Multiphoton; D008954:Models, Biological; D009369:Neoplasms / Q000098:blood supply* / Q000201:enzymology / Q000473:pathology*; D009389:Neovascularization, Pathologic / Q000473:pathology*; D015398:Signal Transduction / Q000187:drug effects; D018874:Spheroids, Cellular / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D017154:Stromal Cells / Q000187:drug effects / Q000473:pathology; D014407:Tumor Cells, Cultured
chemical_list: D020533:Angiogenesis Inhibitors; D036341:Intercellular Signaling Peptides and Proteins; D053511:Matrix Metalloproteinase 14