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Axl-Targeted Cancer Imaging with Humanized Antibody h173

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Molecular Imaging and Biology Aims and scope Submit manuscript

Abstract

Purpose

The tyrosine kinase receptor Axl is overexpressed in various types of cancer and correlated with cancer malignancy. Selective Axl blockade reduces tumor growth and metastasis. The purpose of this study was to examine whether the humanized anti-Axl antibody humanized 173 (h173) labeled with near-infrared fluorescence (NIRF) dye Cy5.5 could be applied as a molecular imaging probe for NIRF imaging of Axl expression in tumor models.

Procedures

NIRF dye Cy5.5 was conjugated to h173 or human normal immunoglobulin G (hIgG) control through amino groups. The resulting probes were evaluated in both A549 (Axl positive) and NCI-H249 (Axl negative) lung cancer xenografts through in vivo NIRF imaging. Ex vivo imaging and probe distribution assay were also carried out to confirm the in vivo imaging results.

Results

After conjugation, binding activity of h173-Cy5.5 was determined to be 97.75 % ± 2.09 % of the unmodified h173. In vitro fluorescence-activated cell sorting (FACS) and fluorescence microscopy analysis validated the specific binding of h173 toward Axl-positive A549 cells. h173-Cy5.5 was then applied to image Axl expression in vivo. In A549 (Axl positive) cancer xenografts, the tumor uptake of h173-Cy5.5 was significantly higher than that of the hIgG-Cy5.5 control (P < 0.05) at late time points (1, 2, 3, 4, and 7 days). On the contrary, in NCI-H249 (Axl negative) cancer xenografts, the tumor uptake of both hIgG-Cy5.5 and h173-Cy5.5 was low and showed no significant difference (P > 0.05) at all time points examined. Ex vivo imaging and immunofluorescence staining analysis further validated the in vivo imaging results.

Conclusions

Collectively, all in vitro, in vivo, and ex vivo data suggested that h173-Cy5.5 could serve as a valid probe for Axl-targeted cancer imaging, which could therefore aid in tumor diagnosis, prognosis, and treatment monitoring.

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Acknowledgments

This work was supported by the NIBIB (R21 1 r21 eb012294-01a1), Early (Margaret E.) Medical Research Trust, the American Cancer Society (121991-MRSG-12-034-01-CCE), SC CTSI (12-2176-3135), Department of Defense (BC102678), Vasgene Therapeutics Inc. (NIH: CA168158-01; CA171538-01), National Natural Science Foundation of China (No. U1032002, 81071206, 81271621, 81301266), and Key Clinical Research Project of Public Health Ministry of China 2010-2012 (No. 164).

Conflict of Interest

The authors declare the following competing financial interest(s): K.N. is the employee of Vasgene Therapeutics Inc.

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Authors and Affiliations

  1. Molecular Imaging Center, Department of Radiology, University of Southern California, 2250 Alcazar St. CSC103, Los Angeles, CA, 90033, USA

    Dan Li, Shuanglong Liu, Ryan Park, Zibo Li & Peter S. Conti

  2. Department of Radiology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510630, China

    Dan Li & Hong Shan

  3. Department of Pathology, University of Southern California, Los Angeles, CA, 90033, USA

    Ren Liu, Haiyang Yu & Parkash S. Gill

  4. National Hepatobiliary and Enteric Surgery Research Center, Department of Surgery, Xiangya Hospital, Central South University, Changsha, 410008, China

    Haiyang Yu

  5. Vasgene Therapeutics Inc., Los Angeles, CA, 90033, USA

    Valery Krasnoperov

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  1. Dan Li
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  2. Shuanglong Liu
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  3. Ren Liu
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Corresponding authors

Correspondence to Zibo Li or Hong Shan.

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Li, D., Liu, S., Liu, R. et al. Axl-Targeted Cancer Imaging with Humanized Antibody h173. Mol Imaging Biol 16, 511–518 (2014). https://doi.org/10.1007/s11307-013-0714-z

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  • DOI: https://doi.org/10.1007/s11307-013-0714-z

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