European Radiology

, Volume 27, Issue 7, pp 2894–2902 | Cite as

Quantification of antiangiogenic treatment effects on tissue heterogeneity in glioma tumour xenograft model using a combination of DCE-MRI and 3D-ultramicroscopy

  • Marco Dominietto
  • Michael Dobosz
  • Sandra Bürgi
  • Anja Renner
  • Gudrun Zahlmann
  • Werner Scheuer
  • Markus Rudin



This study aimed at assessing the effects of an anti-angiogenic treatment, which neutralises vascular endothelial growth factor (VEGF), on tumour heterogeneity.


Murine glioma cells have been inoculated into the right brain frontal lobe of 16 mice. Anti-VEGF antibody was administered to a first group (n = 8), while a second group (n = 8) received a placebo. Magnetic resonance acquisitions, performed at days 10, 12, 15 and 23 following the implantation, allowed the derivation of a three-dimensional features dataset characterising tumour heterogeneity. Three-dimensional ultramicroscopy and standard histochemistry analysis have been performed to verify in vivo results.


Placebo-treated mice displayed a highly-vascularised area at the tumour periphery, a monolithic necrotic core and a chaotic dense vasculature across the entire tumour. In contrast, the B20-treated group did not show any highly vascularised regions and presents a fragmented necrotic core. A significant reduction of the number of vessel segments smaller than 17 μm has been observed. There was no difference in overall tumour volume and growth rate between the two groups.


Region-specific analysis revealed that VEGF inhibition affects only: (1) highly angiogenic compartments expressing high levels of VEGF and characterised by small capillaries, and also (2) the formation and structure of necrotic regions. These effects appear to be transient and limited in time.

Key Points

VEGF inhibition affects only the highly angiogenic region and small capillaries network

VEGF inhibition is transient in time

Tumour volume is not affected by anti-angiogenic treatment

VEGF inhibition also influences the architecture of necrotic regions


Tumour heterogeneity Anti-angiogenic treatment VEGF inhibition Glioma DCE-MRI 



Epithelial growth factor


Fibroblast growth factor


Vascular endothelial growth factor


Mouse glioma cells 261


Vascular permeability (transfer constant)


Vascular leakage space




Murine anti-VEGF monoclonal antibody


Platelet-derived growth factor


Standard error of the mean



The scientific guarantor of this publication is Prof. Markus Rudin. The authors of this manuscript declare relationships with the following companies: F. Hoffmann-La Roche Ltd. The authors state that this work has not received any funding. One of the authors has significant statistical expertise. Approval from the institutional animal care committee was obtained. Methodology: prospective, observational/experimental, performed at two institutions.


  1. 1.
    Burrell RA, McGranahan N, Bartek J, Swanton C (2013) The causes and consequences of genetic heterogeneity in cancer evolution. Nature 501:338–345CrossRefPubMedGoogle Scholar
  2. 2.
    Bedard PL, Hansen AR, Ratain MJ, Siu LL (2013) Tumour heterogeneity in the clinic. Nature 501:355–364CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Dominietto M, Lehmann S, Keist R, Rudin M (2013) Pattern analysis accounts for heterogeneity observed in MRI studies of tumor angiogenesis. Magn Reson Med 70:1481–1490CrossRefPubMedGoogle Scholar
  4. 4.
    Bergers G, Benjamin LE (2003) Angiogenesis: tumorigenesis and the angiogenic switch. Nat Rev Cancer 3:401–410CrossRefPubMedGoogle Scholar
  5. 5.
    Naumov GN, Akslen LA, Folkman J (2006) Role of angiogenesis in human tumor dormancy: animal models of the angiogenic switch. Cell Cycle 5:1779–1787CrossRefPubMedGoogle Scholar
  6. 6.
    Baeriswyl V, Christofori G (2009) The angiogenic switch in carcinogenesis. Semin Cancer Biol 19:329–337CrossRefPubMedGoogle Scholar
  7. 7.
    Folkman J (2007) Angiogenesis: an organizing principle for drug discovery? Nat Rev Drug Discov 6:273–286CrossRefPubMedGoogle Scholar
  8. 8.
    Nagy JA, Dvorak HF (2012) Heterogeneity of the tumor vasculature: the need for new tumor blood vessel type-specific targets. Clin Exp Metastasis 29:657–662CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Fisher R, Pusztai L, Swanton C (2013) Cancer heterogeneity: implications for targeted therapeutics. Br J Cancer 108:479–485CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Dominietto M, Rudin M (2014) Could magnetic resonance provide in vivo histology? Front Genet 4:298CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Young GS (2007) Advanced MRI of adult brain tumors. Neurol Clin 25:947CrossRefPubMedGoogle Scholar
  12. 12.
    Dobosz M, Ntziachristos V, Scheuer W, Strobel S (2014) Multispectral fluorescence ultramicroscopy: three-dimensional visualization and automatic quantification of tumor morphology, drug penetration, and antiangiogenic treatment response. Neoplasia 16:1–13CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Rangayyan RM (2004) Biomedical image analysis. CRC Press, Boca RatonCrossRefGoogle Scholar
  14. 14.
    Zacharaki EI, Wang S, Chawla S et al (2009) Classification of brain tumor type and grade using MRI texture and shape in a machine learning scheme. Magn Reson Med 62:1609–1618CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Theodoridis S, Koutroumbas K (2008) Pattern recognition. Academic Press, BurlingtonGoogle Scholar
  16. 16.
    Chowdhary S, Chamberlain M (2013) Bevacizumab for the treatment of glioblastoma. Expert Rev Neurother 13:937–949CrossRefPubMedGoogle Scholar
  17. 17.
    Curry RC, Dahiya S, Alva Venur V et al (2015) Bevacizumab in high-grade gliomas: past, present, and future. Expert Rev Anticancer Ther 15:387–397CrossRefPubMedGoogle Scholar
  18. 18.
    Vokes EE, Salgia R, Karrison TG (2013) Evidence-based role of bevacizumab in non-small cell lung cancer. Ann Oncol 24:6–9CrossRefPubMedGoogle Scholar
  19. 19.
    Escudier B, Pluzanska A, Koralewski P et al (2007) Bevacizumab plus interferon alfa-2a for treatment of metastatic renal cell carcinoma: a randomised, double-blind phase III trial. Lancet 370:2103–2111CrossRefPubMedGoogle Scholar
  20. 20.
    Saltz LB, Clarke S, Díaz-Rubio E et al (2008) Bevacizumab in combination with oxaliplatin-based chemotherapy as first-line therapy in metastatic colorectal cancer: a randomized phase III study. J Clin Oncol 26:2013–2019CrossRefPubMedGoogle Scholar
  21. 21.
    Soda Y, Myskiw C, Rommel A, Verma IM (2013) Mechanisms of neovascularization and resistance to anti-angiogenic therapies in glioblastoma multiforme. J Mol Med 91:439–448CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Gerstner ER, Batchelor TT (2012) Antiangiogenic therapy for glioblastoma. Cancer J 18:45–50CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Arrillaga-Romany I, Norden AD (2014) Antiangiogenic therapies for glioblastoma. CNS Oncol 3:349–358CrossRefPubMedGoogle Scholar
  24. 24.
    Liang W-C, Wu X, Peale FV et al (2006) Cross-species vascular endothelial growth factor (VEGF)-blocking antibodies completely inhibit the growth of human tumor xenografts and measure the contribution of stromal VEGF. J Biol Chem 281:951–961CrossRefPubMedGoogle Scholar
  25. 25.
    Carano RAD, Ross AL, Ross J et al (2004) Quantification of tumor tissue populations by multispectral analysis. Magn Reson Med 51:542–551CrossRefPubMedGoogle Scholar
  26. 26.
    Tofts PS (1997) Modeling tracer kinetics in dynamic Gd-DTPA MR imaging. J Magn Reson Imaging 7:91–101CrossRefPubMedGoogle Scholar
  27. 27.
    Tofts PS, Brix G, Buckley DL et al (1999) Estimating kinetic parameters from dynamic contrast-enhanced T(1)-weighted MRI of a diffusable tracer: standardized quantities and symbols. J Magn Reson Imaging 10:223–232CrossRefPubMedGoogle Scholar
  28. 28.
    Tofts PS, Kermode AG (1991) Measurement of the blood-brain barrier permeability and leakage space using dynamic MR imaging. 1. Fundamental concepts. Magn Reson Med 17:357–367CrossRefPubMedGoogle Scholar
  29. 29.
    Rudin M, McSheehy PMJ, Allegrini PR et al (2005) PTK787/ZK222584, a tyrosine kinase inhibitor of vascular endothelial growth factor receptor, reduces uptake of the contrast agent GdDOTA by murine orthotopic B16/BL6 melanoma tumours and inhibits their growth in vivo. NMR Biomed 18:308–321CrossRefPubMedGoogle Scholar
  30. 30.
    Allain C, Cloitre M (1991) Characterizing the lacunarity of random and deterministic fractal sets. Phys Rev A 44:3552–3558CrossRefPubMedGoogle Scholar
  31. 31.
    Guyon I, Elisseeff A (2003) An introduction to variable and feature selection. J Mach Learn Res 3:1157–1182Google Scholar
  32. 32.
    Umbaugh SE (2010) Digital image processing and analysis: human and computer vision applications with CVIPtools, 2nd edn. CRC Press, Boca RatonGoogle Scholar
  33. 33.
    Folkman J (1971) Tumor angiogenesis: therapeutic implications. N Engl J Med 285:1182–1186CrossRefPubMedGoogle Scholar
  34. 34.
    Goel S, Duda DG, Xu L et al (2011) Normalization of the vasculature for treatment of cancer and other diseases. Physiol Rev 91:1071–1121CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Jain RK (2005) Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science 307:58–62CrossRefPubMedGoogle Scholar
  36. 36.
    Dickson PV, Hamner JB, Sims TL et al (2007) Revacizumab-induced transient remodeling of the vasculature in neuroblastoma xenografts results in improved delivery and efficacy of systemically administered chemotherapy. Clin Cancer Res 13:3942–3950CrossRefPubMedGoogle Scholar
  37. 37.
    Kepes JJ (2003) Necrosis and glioblastoma: a friend or a foe? A review and a hypothesis. Neurosurgery 52:1242PubMedGoogle Scholar

Copyright information

© European Society of Radiology 2016

Authors and Affiliations

  1. 1.Institute for Biomedical EngineeringUniversity and ETH ZurichZurichSwitzerland
  2. 2.Biomaterials Science CenterUniversity of BaselAllschwilSwitzerland
  3. 3.Discovery Oncology, Pharmaceutical Research and Early Development (pRED)Roche Innovation Center PenzbergPenzbergGermany
  4. 4.pRED, Oncology DTA, Innovation Center Basel, F. Hoffmann-La Roche LtdBaselSwitzerland

Personalised recommendations