The cancer angiogenesis co-culture assay: In vitro quantification of the angiogenic potential of tumoroids

PLoS One. 2021 Jul 7;16(7):e0253258. doi: 10.1371/journal.pone.0253258. eCollection 2021.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • Angiogenesis Inducing Agents / pharmacology
  • Angiogenesis Inhibitors / pharmacology*
  • Cell Line, Tumor
  • Coculture Techniques / methods*
  • Colorectal Neoplasms / drug therapy
  • Colorectal Neoplasms / pathology
  • Female
  • Fibroblast Growth Factors / pharmacology
  • Fibroblasts / drug effects
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Humans
  • In Vitro Techniques
  • Male
  • Middle Aged
  • Neovascularization, Pathologic / drug therapy*
  • Platelet-Derived Growth Factor / pharmacology
  • Spheroids, Cellular / drug effects
  • Vascular Endothelial Growth Factor A / pharmacology

Substances

  • Angiogenesis Inducing Agents
  • Angiogenesis Inhibitors
  • Platelet-Derived Growth Factor
  • Vascular Endothelial Growth Factor A
  • platelet-derived growth factor A
  • Fibroblast Growth Factors