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In vivo near-infrared fluorescence imaging of CD105 expression during tumor angiogenesis

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European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

Angiogenesis is an indispensable process during tumor development. The currently accepted standard method for quantifying tumor angiogenesis is to assess microvessel density (MVD) based on CD105 staining, which is an independent prognostic factor for survival in patients with most solid tumor types. The goal of this study is to evaluate tumor angiogenesis in a mouse model by near-infrared fluorescence (NIRF) imaging of CD105 expression.

Methods

TRC105, a human/murine chimeric anti-CD105 monoclonal antibody, was conjugated to an NIRF dye (IRDye 800CW; Ex: 778 nm; Em: 806 nm). FACS analysis and microscopy studies were performed to compare the CD105 binding affinity of TRC105 and 800CW-TRC105. In vivo/ex vivo NIRF imaging, blocking studies, and ex vivo histology were performed on 4T1 murine breast tumor-bearing mice to evaluate the ability of 800CW-TRC105 to target tumor angiogenesis. Another chimeric antibody, cetuximab, was used as an isotype-matched control.

Results

FACS analysis of human umbilical vein endothelial cells (HUVECs) revealed no difference in CD105 binding affinity between TRC105 and 800CW-TRC105, which was further validated by fluorescence microscopy. 800CW conjugation of TRC105 was achieved in excellent yield (> 85%), with an average of 0.4 800CW molecules per TRC105. Serial NIRF imaging after intravenous injection of 800CW-TRC105 revealed that the 4T1 tumor could be clearly visualized as early as 30 min post-injection. Quantitative region of interest (ROI) analysis showed that the tumor uptake peaked at about 16 h post-injection. Based on ex vivo NIRF imaging at 48 h post-injection, tumor uptake of 800CW-TRC105 was higher than most organs, thus providing excellent tumor contrast. Blocking experiments, control studies with 800CW-cetuximab and 800CW, as well as ex vivo histology all confirmed the in vivo target specificity of 800CW-TRC105.

Conclusion

This is the first successful NIRF imaging study of CD105 expression in vivo. Fast, prominent, persistent, and CD105-specific uptake of the probe during tumor angiogenesis was observed in a mouse model. 800CW-TRC105 may be used in the clinic for imaging tumor angiogenesis within the lesions close to the skin surface, tissues accessible by endoscopy, or during image-guided surgery.

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Acknowledgements

This work is supported, in part, by the University of Wisconsin Carbone Cancer Center, NCRR 1UL1RR025011, a DOD BCRP Postdoctoral Fellowship, a Susan G. Komen Postdoctoral Fellowship, and a DOD PCRP IDEA Award.

Conflicts of interest

BRL is an employee of TRACON Pharmaceuticals, Inc. The other authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Radiology, University of Wisconsin-Madison, Madison, WI, USA

    Yunan Yang, Hao Hong & Weibo Cai

  2. Department of Ultrasound, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, People’s Republic of China

    Yunan Yang

  3. Department of Medical Physics, University of Wisconsin-Madison, Madison, WI, USA

    Yin Zhang & Weibo Cai

  4. University of Wisconsin Carbone Cancer Center, Madison, WI, USA

    Glenn Liu & Weibo Cai

  5. TRACON Pharmaceuticals, Inc., San Diego, CA, USA

    Bryan R. Leigh

  6. Departments of Radiology and Medical Physics, School of Medicine and Public Health, University of Wisconsin-Madison, 1111 Highland Ave, Room 7137, Madison, WI, 53705-2275, USA

    Weibo Cai

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  1. Yunan Yang
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  3. Hao Hong
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Correspondence to Weibo Cai.

Additional information

Yunan Yang and Yin Zhang contributed equally to this work.

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Yang, Y., Zhang, Y., Hong, H. et al. In vivo near-infrared fluorescence imaging of CD105 expression during tumor angiogenesis. Eur J Nucl Med Mol Imaging 38, 2066–2076 (2011). https://doi.org/10.1007/s00259-011-1886-x

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  • DOI: https://doi.org/10.1007/s00259-011-1886-x

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