Abstract
The treatment response to anti-angiogenic agents varies among cancer patients and predictive biomarkers are needed to identify patients with resistant cancer or guide the choice of anti-angiogenic treatment. We present "the Cancer Angiogenesis Co-Culture (CACC) assay", an in vitro Functional Precision Medicine assay which enables the study of tumouroid induced angiogenesis. This assay can quantify the ability of a patient-derived tumouroid to induce vascularization by measuring the induction of tube formation in a co-culture of vascular cells and tumoroids established from the primary colorectal tumour or a metastasis. Furthermore, the assay can quantify the sensitivity of patient-derived tumoroids to anti-angiogenic therapies. We observed that tube formation increased in a dose-dependent manner upon treatment with the pro-angiogenic factor vascular endothelial growth factor A (VEGF-A). When investigating the angiogenic potential of tumoroids from 12 patients we found that 9 tumoroid cultures induced a significant increase in tube formation compared to controls without tumoroids. In these 9 angiogenic tumoroid cultures the tube formation could be abolished by treatment with one or more of the investigated anti-angiogenic agents. The 3 non-angiogenic tumoroid cultures secreted VEGF-A but we observed no correlation between the amount of tube formation and tumoroid-secreted VEGF-A. Our data suggests that the CACC assay recapitulates the complexity of tumour angiogenesis, and when clinically verified, could prove a valuable tool to quantify sensitivity towards different anti-angiogenic agents.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aged
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Aged, 80 and over
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Angiogenesis Inducing Agents / pharmacology
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Angiogenesis Inhibitors / pharmacology*
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Cell Line, Tumor
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Coculture Techniques / methods*
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Colorectal Neoplasms / drug therapy
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Colorectal Neoplasms / pathology
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Female
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Fibroblast Growth Factors / pharmacology
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Fibroblasts / drug effects
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Human Umbilical Vein Endothelial Cells / drug effects
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Humans
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In Vitro Techniques
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Male
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Middle Aged
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Neovascularization, Pathologic / drug therapy*
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Platelet-Derived Growth Factor / pharmacology
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Spheroids, Cellular / drug effects
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Vascular Endothelial Growth Factor A / pharmacology
Substances
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Angiogenesis Inducing Agents
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Angiogenesis Inhibitors
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Platelet-Derived Growth Factor
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Vascular Endothelial Growth Factor A
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platelet-derived growth factor A
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Fibroblast Growth Factors
Grants and funding
SLBT and NM was supported by the Innovation Fond Denmark
http://innovationsfonden.dk/en), grant no. 5184-00101B. SLBT received funding from Agnes & Poul Friis Fund (grant no. 81008-001), C.C. Klestrup & hustrus Henriette Klestrups Mindelegat (grant no. 0660-001), Carl & Ellen Hertz legat til dansk læge- og naturvidenskab (grant no. 7179-2), The Drost Fundation (grant no. 12120-1), and Familien Hede Nielsens Fund (grant no 2017-0423). The funders provided support in the form of salaries for authors SLBT and NM but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. SLBT, GH, OT, RS and JT are fulltime employees at 2cureX, Symbion, Copenhagen, Denmark (
https://www.2curex.com/). 2cureX provided support in the form of salaries for authors GH, OT, RS and JT. ES is a fulltime employee at BioSense Solutions, Farum, Denmark (
https://biosensesolutions.dk/). BioSense Solutions provided support in the form of salaries for ES. HW, LH, KQ, HH and HM received no specific funding for this work.