Abstract
The sensitivity of Doppler ultrasound below 10 MHz to assess antiangiogenic therapy effects in tumor xenografts has been shown to be limited. Thus, our aim was to evaluate high-frequency volumetric power-Doppler ultrasound (HF-VPDU) for monitoring antiangiogenic treatments. Squamous cell carcinoma xenografts grown in nude mice were scanned with HF-VPDU at a center frequency of 30 MHz. Images with 200-μm slice thicknesses were recorded and merged into a three-dimensional dataset, of which the relative color pixel density was determined. Animals received either VEGFR2 antibodies or 0.9% NaCl and were examined at days 0, 3 and 6 of treatment. After the last examination, tumors were resected and their vascularization characterized by immunohistology. At day 6, the volumes of treated and untreated tumors were not significantly different yet. In contrast, mean tumor vascularization in treated animals had decreased to 44%, while in the control group it had increased to 152% (P < 0.01). In correspondence, vessel density, as determined by CD31 staining, was 0.19 ± 0.10% in treated and 0.92 ± 0.41% in untreated tumors (P < 0.01). Additionally, the fraction of mature (SMA-positive) vessels increased under therapy. Thus, HF-VPDU can be considered as an easily applicable and fast method to screen high animal numbers for antiangiogenic therapy effects.
Similar content being viewed by others
References
Folkman J (2006) Angiogenesis. Annu Rev Med 57:1–18
Folkman J (2003) Angiogenesis inhibitors: a new class of drugs. Cancer Biol Ther 2:127–133
Ferrara N, Kerbel RS (2005) Angiogenesis as a therapeutic target. Nature 15:967–974
Kiessling F, Farhan N, Lichy MP, Vosseler S, Heilmann M, Krix M, Bohlen P, Miller DW, Mueller MM, Semmler W, Fusenig NE, Delorme S (2004) Dynamic contrast-enhanced magnetic resonance imaging rapidly indicates vessel regression in human squamous cell carcinomas grown in nude mice caused by VEGF receptor 2 blockade with DC101. Neoplasia 6:213–223
Fleischer AC, Niermann KJ, Donnelly EF, Yankeelov TE, Canniff KM, Hallahan DE, Rothenberg ME (2004) Sonographic depiction of microvessel perfusion: principles and potential. J Ultrasound Med 23:1499–1506
Neeman M, Gilad AA, Dafni H, Cohen B (2007) Molecular imaging of angiogenesis. J Magn Reson Imaging 25:1–12
Fleischer AC, Milam MR, Crispens MA, Shappell HW (2005) Sonographic depiction of intratumoral vascularity with 2- and 3-dimensional color Doppler techniques. J Ultrasound Med 24:533–537
Krix M, Kiessling F, Vosseler S, Farhan N, Mueller MM, Bohlen P, Fusenig NE, Delorme S (2003) Sensitive noninvasive monitoring of tumor perfusion during antiangiogenic therapy by intermittent bolus-contrast power Doppler sonography. Cancer Res 63:8264–8270
Lee TY, Purdie TG, Stewart E (2003) CT imaging of angiogenesis. Q J Nucl Med 47:171–187
Laking GR, Price PM (2003) Positron emission tomographic imaging of angiogenesis and vascular function. Br J Radiol 76:50–59
Delorme S, Krix M (2006) Contrast-enhanced ultrasound for examining tumor biology. Cancer Imaging 6:148–152
Lindner JR (2004) Microbubbles in medical imaging: current applications and future directions. Nat Rev Drug Discov 3:527–532
Qin S, Ferrara KW (2006) Acoustic response of compliable microvessels containing ultrasound contrast agents. Phys Med Biol 51:5065–5088
Goertz DE, Christopher DA, Yu JL, Kerbel RS, Burns PN, Foster FS (2000) High-frequency color flow imaging of the microcirculation. Ultrasound Med Biol 26:63–71
Fusenig NE, Boukamp P (1998) Multiple stages and genetic alterations in immortalization, malignant transformation, and tumor progression of human skin keratinocytes. J Mol Carcinogenesis 23:144–158
Mueller MM, Peter W, Mappes M, Huelsen A, Steinbauer H, Boukamp P, Vaccariello M, Garlick J, Fusenig NE (2001) Tumor progression of skin carcinoma cells in vivo promoted by clonal selection, mutagenesis, and autocrine growth regulation by granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. Am J Pathol 159:1567–1579
Gee MS, Saunders HM, Lee JC, Sanzo JF, Jenkins WT, Evans SM, Trinchieri G, Sehgal CM, Feldman MD, Lee WM (2001) Doppler ultrasound imaging detects changes in tumor perfusion during antivascular therapy associated with vascular anatomic alterations. Cancer Res 61:2974–2982
Goertz DE, Yu JL, Kerbel RS, Burns PN, Foster FS (2002) High-frequency Doppler ultrasound monitors the effects of antivascular therapy on tumor blood flow. Cancer Res 62:6371–6375
Rubin JM, Fowlkes JB, Tuthill TA, Moskalik AP, Rhee RT, Adler RS, Kazanjian SN, Carson PL (1999) Speckle decorrelation flow measurement with B-mode US of contrast agent flow in a phantom and in rabbit kidney. Radiology 213:429–437
Brown EB, Campbell RB, Tsuzuki Y, Xu L, Carmeliet P, Fukumura D, Jain RK (2001) In vivo measurement of gene expression, angiogenesis and physiological function in tumors using multiphoton laser scanning microscopy. Nat Med 7:864–868
Cheung AM, Brown AS, Cucevic V, Roy M, Needles A, Yang V, Hicklin DJ, Kerbel RS, Foster FS (2007) Detecting vascular changes in tumor xenografts using micro-ultrasound and micro-ct following treatment with VEGFR-2 blocking antibodies. Ultrasound Med Biol 8:1259–1268
Jain RK (2003) Molecular regulation of vessel maturation. Nat Med 9:685–693
Acknowledgements
This work was supported by the transregional grant Vascular differentiation and remodeling from the German Research Foundation (DFG, SFB-TR23).
Author information
Authors and Affiliations
Corresponding author
Additional information
M. Jugold and M. Palmowski contributed equally to this work
Rights and permissions
About this article
Cite this article
Jugold, M., Palmowski, M., Huppert, J. et al. Volumetric high-frequency Doppler ultrasound enables the assessment of early antiangiogenic therapy effects on tumor xenografts in nude mice. Eur Radiol 18, 753–758 (2008). https://doi.org/10.1007/s00330-007-0825-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00330-007-0825-5