Advertisement

European Radiology

, Volume 21, Issue 9, pp 1988–1995 | Cite as

Molecular and functional ultrasound imaging in differently aggressive breast cancer xenografts using two novel ultrasound contrast agents (BR55 and BR38)

  • Jessica Bzyl
  • Wiltrud Lederle
  • Anne Rix
  • Christoph Grouls
  • Isabelle Tardy
  • Sibylle Pochon
  • Monica Siepmann
  • Tobias Penzkofer
  • Michel Schneider
  • Fabian Kiessling
  • Moritz Palmowski
Molecular Imaging

Abstract

Objectives

To characterise clinically translatable long-circulating (BR38) and VEGFR2-targeted (BR55) microbubbles (MB) and to assess their ability to discriminate breast cancer models with different aggressiveness.

Methods

The circulation characteristics of BR38 and BR55 were investigated in healthy mice. The relative blood volume (rBV) of MDA-MB-231 (n = 5) or MCF-7 (n = 6) tumours was determined using BR38. In the same tumours in-vivo binding specificity of BR55 was tested and VEGFR2 expression assessed. Data validation included quantitative immunohistological analysis.

Results

BR38 had a longer blood half-life than BR55 (>600 s vs. 218 s). BR38-enhanced ultrasound showed greater vascularisation in MDA-MB-231 tumours (p = 0.022), which was in line with immunohistology (p = 0.033). In-vivo competitive binding experiments proved the specificity of BR55 to VEGFR2 (p = 0.027). Binding of BR55 was significantly higher in MDA-MB-231 than in MCF-7 tumours (p = 0.049), which corresponded with the VEGFR2 levels found histologically (p = 0.015). However, differences became smaller when normalising the levels of BR55 to the rBV.

Conclusions

BR38 and BR55 are well suited to characterising and distinguishing breast cancers with different angiogenesis and aggressiveness. Long-circulating BR38 MB allow extensive 3-dimensional examinations of larger or several organs. BR55 accumulation faithfully reflects the VEGFR2 status in tumours and depicts even small differences in angiogenesis.

Keywords

Functional and molecular ultrasound Angiogenesis VEGFR2 Breast cancer 

References

  1. 1.
    Kaiser WA (2007) Breast magnetic resonance imaging: principles and techniques. Semin Roentgenol 42:228–235PubMedCrossRefGoogle Scholar
  2. 2.
    Kiessling F, Huppert J, Palmowski M (2009) Functional and molecular ultrasound imaging: concepts and contrast agents. Curr Med Chem 16:627–642PubMedCrossRefGoogle Scholar
  3. 3.
    Berg WA, Blume JD, Cormack JB et al (2008) Combined screening with ultrasound and mammography vs mammography alone in women at elevated risk of breast cancer. JAMA 299:2151–2163PubMedCrossRefGoogle Scholar
  4. 4.
    Balleyguier C, Opolon P, Mathieu MC et al (2009) New potential and applications of contrast-enhanced ultrasound of the breast: own investigations and review of the literature. Eur J Radiol 69:14–23PubMedCrossRefGoogle Scholar
  5. 5.
    Palmowski M, Huppert J, Hauff P et al (2008) Vessel fractions in tumor xenografts depicted by flow- or contrast-sensitive three-dimensional high-frequency Doppler ultrasound respond differently to antiangiogenic treatment. Cancer Res 68:7042–7049PubMedCrossRefGoogle Scholar
  6. 6.
    Palmowski M, Lederle W, Gaetjens J et al (2010) Comparison of conventional time-intensity curves vs. maximum intensity over time for post processing of dynamic contrast-enhanced ultrasound. Eur J Radiol 75:149–153CrossRefGoogle Scholar
  7. 7.
    Klibanov AL (2005) Ligand-carrying gas-filled microbubbles: ultrasound contrast agents for targeted molecular imaging. Bioconjug Chem 16:9–17PubMedCrossRefGoogle Scholar
  8. 8.
    Lindner JR (2004) Microbubbles in medical imaging: current applications and future directions. Nat Rev Drug Discov 3:527–532PubMedCrossRefGoogle Scholar
  9. 9.
    Dayton PA, Rychak JJ (2007) Molecular ultrasound imaging using microbubble contrast agents. Front Biosci 12:5124–5142PubMedCrossRefGoogle Scholar
  10. 10.
    Shibuya M (2006) Differential roles of vascular endothelial growth factor receptor-1 and receptor-2 in angiogenesis. J Biochem Mol Biol 39:469–478PubMedCrossRefGoogle Scholar
  11. 11.
    Willmann JK, Paulmurugan R, Chen K et al (2008) US imaging of tumor angiogenesis with microbubbles targeted to vascular endothelial growth factor receptor type 2 in mice. Radiology 246:508–518PubMedCrossRefGoogle Scholar
  12. 12.
    Willmann JK, Lutz AM, Paulmurugan R (2008) Dual-targeted contrast agent for US assessment of tumor angiogenesis in vivo. Radiology 248:936–944PubMedCrossRefGoogle Scholar
  13. 13.
    Lyshick A, Fleischer AC, Huamani J et al (2007) Molecular imaging of vascular endothelial growth factor receptor 2 expression using targeted contrast-enhanced high-frequency ultrasonography. J Ultrasound Med 26:1575–1586Google Scholar
  14. 14.
    Palmowski M, Morgenstern B, Hauff P et al (2008) Pharmacodynamics of streptavidin-coated cyanoacrylate microbubbles designed for molecular ultrasound imaging. Invest Radiol 43:162–169PubMedCrossRefGoogle Scholar
  15. 15.
    Palmowski M, Huppert J, Ladewig G et al (2008) Molecular profiling of angiogenesis with targeted ultrasound imaging: early assessment of anti-angiogenic therapy effects. Mol Cancer Ther 7:101–109PubMedCrossRefGoogle Scholar
  16. 16.
    Rychak JJ, Graba J, Cheung AM et al (2007) Microultrasound molecular imaging of vascular endothelial growth factor receptor 2 in a mouse model of tumor angiogenesis. Mol Imaging 6:289–296PubMedGoogle Scholar
  17. 17.
    Korpanty G, Carbon JG, Grayburn PA et al (2007) Monitoring response to anticancer therapy by targeting microbubbles to tumor vasculature. Clin Cancer Res 13:323–330PubMedCrossRefGoogle Scholar
  18. 18.
    Pochon S, Tardy I, Bussat P et al (2010) BR55: A lipopeptide-based VEGFR2-targeted ultrasound contrast agent for molecular imaging of angiogenesis. Invest Radiol 45:89–95PubMedCrossRefGoogle Scholar
  19. 19.
    Pysz MA, Foygel K, Rosenberg J et al (2010) Antiangiogenic cancer therapy: monitoring with molecular US and a clinically translatable contrast agent (BR55). Radiology 256:519–527PubMedCrossRefGoogle Scholar
  20. 20.
    Naik MU, Naik TU, Suckow AT et al (2008) Attenuation of junctional adhesion molecule-A is a contributing factor for breast cancer cell invasion. Cancer Res 68:2194–2203PubMedCrossRefGoogle Scholar
  21. 21.
    Wang C, Navab R, Iakovlev V et al (2007) Abelson interactor protein-1 positively regulates breast cancer cell proliferation, migration, and invasion. Mol Cancer Res 5:1031–1039PubMedCrossRefGoogle Scholar
  22. 22.
    Kasper G, Reule M, Tschirschmann M et al (2007) Stromelysin-3 over-expression enhances tumourigenesis in MCF-7 and MDA-MB-231 breast cancer cell lines: involvement of the IGF-1 signalling pathway. BMC Cancer 17:7–12Google Scholar
  23. 23.
    Shrivastava A, von Wronski MA, Sato AK et al (2005) A distinct strategy to generate high-affinity peptide binders to receptor tyrosine kinases. Protein Eng Des Sel 18:417–424PubMedCrossRefGoogle Scholar
  24. 24.
    Pillai R, Marinelli ER, Swenson RE (2006) A Flexible Method for preparation of peptide homo- and heterodimers functionalized with affinity probes, chelating ligands, and latent conjugating groups. Biopolymers (Peptide Sci) 84:576–585CrossRefGoogle Scholar
  25. 25.
    Pillai R, Marinelli ER, Fan H (2010) A phospholipid-PEG2000 conjugate of a vascular endothelial growth factor receptor 2 (VEGFR2)-targeting heterodimer peptide for contrast-enhanced ultrasound imaging of angiogenesis. Bioconjug Chem 21:556–562CrossRefGoogle Scholar
  26. 26.
    Watanabe R, Matsumura M, Munemasa T et al (2007) Mechanism of hepatic parenchyma-specific contrast of microbubble-based contrast agent for ultrasonography: microscopic studies in rat liver. Invest Radiol 42:643–651PubMedCrossRefGoogle Scholar
  27. 27.
    Morel DR, Schwieger I, Hohn L et al (2000) Human pharmacokinetics and safety evaluation of SonoVue, a new contrast agent for ultrasound imaging. Invest Radiol 35:80–85PubMedCrossRefGoogle Scholar
  28. 28.
    Lindner JR, Dayton PA, Coggins MP (2000) Noninvasive imaging of inflammation by ultrasound detection of phagocytosed microbubbles. Circulation 102:531–538PubMedGoogle Scholar
  29. 29.
    Ferrara KW, Borden MA, Zhang H (2009) Lipid shelled vehicles: engineering for ultrasound molecular imaging and drug delivery. Acc Chem Res 42:881–892PubMedCrossRefGoogle Scholar
  30. 30.
    Wei K, Jayaweera AR, Firoozan S et al (1998) Quantification of myocardial blood flow with ultrasound induced destruction of microbubbles administered as a constant venous infusion. Circulation 97:473–483PubMedGoogle Scholar
  31. 31.
    Krix M, Kiessling F, Vosseler S et al (2003) Sensitive noninvasive monitoring of tumor perfusion during antiangiogenic therapy by intermittent bolus-contrast power Doppler sonography. Cancer Res 63:8264–8279PubMedGoogle Scholar
  32. 32.
    Shimizu H, Miyazaki M, Wakabayashi Y et al (2001) Vascular endothelial growth factor secreted by replicating hepatocytes induces sinusoidal endothelial cell proliferation during regeneration after partial hepatectomy in rats. J Hepatol 34:683–689PubMedCrossRefGoogle Scholar
  33. 33.
    Willmann JK, Cheng Z, Davis C et al (2008) Targeted microbubbles for imaging tumor angiogenesis: assessment of whole-body biodistribution with dynamic micro-PET in mice. Radiology 249:212–219PubMedCrossRefGoogle Scholar
  34. 34.
    Dadiani M, Kalchenko V, Yosepovich A (2006) Real-time imaging of lymphangiogenic metastasis in orthotopic human breast cancer. Cancer Res 66:8037–8041PubMedCrossRefGoogle Scholar
  35. 35.
    Hicklin DJ, Ellis LM (2005) Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis. J Clin Oncol 23:1011–1027PubMedCrossRefGoogle Scholar

Copyright information

© European Society of Radiology 2011

Authors and Affiliations

  • Jessica Bzyl
    • 1
  • Wiltrud Lederle
    • 1
  • Anne Rix
    • 1
  • Christoph Grouls
    • 1
    • 2
  • Isabelle Tardy
    • 3
  • Sibylle Pochon
    • 3
  • Monica Siepmann
    • 4
  • Tobias Penzkofer
    • 2
  • Michel Schneider
    • 3
  • Fabian Kiessling
    • 1
  • Moritz Palmowski
    • 1
    • 2
    • 5
  1. 1.Institute for Experimental Molecular ImagingRWTH-Aachen UniversityAachenGermany
  2. 2.Department of RadiologyRWTH-Aachen UniversityAachenGermany
  3. 3.Bracco Research SAGenevaSwitzerland
  4. 4.Institute of Medical EngineeringRuhr-University BochumBochumGermany
  5. 5.Department of Nuclear MedicineRWTH-Aachen UniversityAachenGermany

Personalised recommendations