Korean J Radiol.  2010 Aug;11(4):449-456. 10.3348/kjr.2010.11.4.449.

Visualization of Tumor Angiogenesis Using MR Imaging Contrast Agent Gd-DTPA-anti-VEGF Receptor 2 Antibody Conjugate in a Mouse Tumor Model

• Jun HY
• Yin HH
• Kim SH
• Park SH
• Kim HS
• Yoon KH

• KMID: 984899
• DOI: http://doi.org/10.3348/kjr.2010.11.4.449

Abstract

OBJECTIVE
To visualize tumor angiogenesis using the MRI contrast agent, Gd-DTPA-anti-VEGF receptor 2 antibody conjugate, with a 4.7-Tesla MRI instrument in a mouse model.
MATERIALS AND METHODS
We designed a tumor angiogenesis-targeting T1 contrast agent that was prepared by the bioconjugation of gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) and an anti-vascular endothelial growth factor receptor-2 (VEGFR2) antibody. The specific binding of the agent complex to cells that express VEGFR2 was examined in cultured murine endothelial cells (MS-1 cells) with a 4.7-Tesla magnetic resonance imaging scanner. Angiogenesis-specific T1 enhancement was imaged with the Gd-DTPA-anti-VEGFR2 antibody conjugate using a CT-26 adenocarcinoma tumor model in eight mice. As a control, the use of the Gd-DTPA-anti-rat immunoglobulin G (Gd-DTPA-anti-rat IgG) was imaged with a tumor model in eight mice. Statistical significance was assessed using the Mann-Whitney test. Tumor tissue was examined by immunohistochemical analysis.
RESULTS
The Gd-DTPA-anti-VEGFR2 antibody conjugate showed predominant binding to cultured endothelial cells that expressed a high level of VEGFR2. Signal enhancement was approximately three-fold for in vivo T1-weighted MR imaging with the use of the Gd-DTPA-anti-VEGFR2 antibody conjugate as compared with the Gd-DTPA-rat IgG in the mouse tumor model (p < 0.05). VEGFR2 expression in CT-26 tumor vessels was demonstrated using immunohistochemical staining.
CONCLUSION
MR imaging using the Gd-DTPA-anti-VEGFR2 antibody conjugate as a contrast agent is useful in visualizing noninvasively tumor angiogenesis in a murine tumor model.

Keyword

Magnetic resonance (MR) contrast agent; Molecular Imaging; Angiogenesis; Bioconjugation

MeSH Terms

Adenocarcinoma/*pathology
Animals
Colonic Neoplasms/*pathology
Contrast Media/chemistry/*diagnostic use
Gadolinium DTPA/chemistry/*diagnostic use
Immunoenzyme Techniques
Magnetic Resonance Imaging/*methods
Mice
Mice, Nude
Neovascularization, Pathologic/*diagnosis
Rats
Statistics, Nonparametric
Tumor Cells, Cultured
Vascular Endothelial Growth Factor Receptor-2/*antagonists & inhibitors/chemistry

Figure

• 

  Fig. 1 Schematic diagram depicting preparation of Gd-DTPA-anti-VEGFR2 antibody is shown. DTPABA = diethylenetriaminepentaacetic acid bisanhydride, mAb = monoclonal antibody (anti-VEGFR 2 antibody), PBS = phosphate-buffered saline, Gd = gadolinium

• 

  Fig. 2 T1-weighted MR images using Gd-DTPA-anti-VEGFR2 antibody in cultured endothelial cells are shown. A. MS-1 cells were not treated with either contrast or control agent. B. VEGFR2 expressing MS-1 cells treated with Gd-DTPA-anti-VEGFR2 antibody shows significant increase in high signal intensity. C. MS-1 cells show no high signal on T1-weighted imaging with Gd-DTPA-anti-VEGFR2 antibody in presence of competitive binding inhibitor, anti-VEGFR2 antibody.

• 

  Fig. 3 In vivo dynamic MR imaging of tumors is shown at 10 minutes and at 12, 24 and 48 hours following injection of Gd-DTPA-anti-VEGFR2 antibody conjugate (A-E) as well as injection of Gd-DTPA-anti-rat IgG (F-J). Both contrast agents were injected at same dose. Gd-DTPA-anti-VEGFR2 antibody conjugate shows significant signal enhancement and accumulation in tumor tissues until 48 hours after contrast agent injection. Comparatively, Gd-DTPA-anti-rat IgG shows no significant enhancement in T1 until 48 hours after injection.

• 

  Fig. 4 Histopathological examination results. A. Hematoxylin and eosin staining shows tumor tissue. B. Tissue was stained with rabbit anti-mouse VEGFR2 (Flk-1) antibody for immunohistochemical analysis, indicating expression of VEGFR2 in vessels (arrows) of CT-26 tumor tissue (×400 magnification).

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