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Identification of a CEACAM5 targeted nanobody for positron emission tomography imaging and near-infrared fluorescence imaging of colorectal cancer

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Abstract

Purpose

Here, we aim to identify a CEACAM5-targeted nanobody and demonstrate its application in positron emission tomography (PET) imaging and near-infrared (NIR) fluorescence imaging in colorectal cancer (CRC).

Methods

Immunohistochemistry was applied to verify CEACAM5 expression in CRC and metastatic lymph nodes (mLNs). CEACAM5-targeted nanobodies were obtained by immunization of human CEACAM5 protein in a dromedary, followed by several rounds of phage screenings. Immunofluorescence staining and flow cytometry was carried out to determine the binding affinity of the nanobodies. The nanobodies were radiolabeled by coupling 18F-SFB for PET imaging of CRC subcutaneous xenografts and lymph node metastasis (LNM). IRDye800CW (IR800) were conjugated to form NIR probes for NIR imaging in CRC subcutaneous models.

Results

CEACAM5 was overexpressed in either human CRC tissues or mLNs. A CEACAM5 targeted nanobody, Nb41 was successfully generated, with excellent in vitro binding properties. Incorporation of albumin binding domain (ABD) did not affect the affinity of Nb41. In vivo imaging showed that both 18F-FB-Nb41 and 18F-FB-Nb41-ABD showed obvious accumulation in the tumor. Due to the longer retention in the blood, 18F-FB-Nb41-ABD enrichment in tumors was significantly delayed but higher compared to 18F-FB-Nb41. Both 18F-FB-Nb41 and 18F-FB-Nb41-ABD showed prominent LNM enrichment. Similarly, the IR800-conjugated nanobodies Nb41-IR800 and Nb41-ABD-IR800 exhibited superior imaging effects in subcutaneous models, while Nb41-ABD-IR800 exhibited higher fluorescence intensity in the tumor accompanied with a remarkedly delay compared to Nb41-IR800.

Conclusion

Collectively, we presented the identification and in vivo validation of a CEACAM5-targeted nanobody and a fused nanobody with an ABD, which enabled to the non-invasive visualization of malignancy of CRC using PET imaging and NIR imaging in subcutaneous models as well as LNM models.

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Data Availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We appreciate Shaohui Deng and Yuanqiang Xiao for the assistance in the synthesis of precursor compounds. We are grateful to Tianxing Zhu and Jiayi Jiang for technical support of MRI. We would like to express our sincere gratitude to Guolong Huang for his help in PET imaging.

Funding

This work was supported by grants from the National Natural Science Foundation of China (92259204) and the Department of Science and Technology of Guangdong Province to the Guangdong Provincial Key Laboratory of Biomedical Imaging (2018B030322006).

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Author notes

  1. Yitai Xiao, Chaoming Mei and Duo Xu These authors contributed equally.

Authors and Affiliations

  1. Department of Nuclear Medicine, The Fifth Affiliated Hospital, Sun Yat-Sen University, Guangdong Province, 519000, China

    Yitai Xiao, Chaoming Mei, Duo Xu, Fan Yang, Meilin Yang & Dan Li

  2. Guangdong Provincial Key Laboratory of Biomedical Imaging and Guangdong Provincial Engineering Research Center of Molecular Imaging, The Fifth Affiliated Hospital, Sun Yat-Sen University, Guangdong Province, 519000, China

    Yitai Xiao, Chaoming Mei, Duo Xu, Fan Yang, Meilin Yang, Lei Bi & Dan Li

  3. Center for Interventional Medicine, The Fifth Affiliated Hospital, Sun Yat-Sen University, Guangdong Province, 519000, China

    Junjie Mao & Pengfei Pang

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  1. Yitai Xiao
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Contributions

Conception: Dan Li, Pengfei Pang, Junjie Mao. Methodology: Yitai Xiao, Chaoming Mei, Duo Xu, Fan Yang, Meilin Yang. Data acquisition and analysis: Yitai Xiao, Chaoming Mei, Duo Xu, Lei Bi. Manuscript preparation and revision: Yitai Xiao, Chaoming Mei, Duo Xu. Study supervision: Dan Li.

Corresponding authors

Correspondence to Junjie Mao, Pengfei Pang or Dan Li.

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The authors have no relevant financial or non-financial interests to disclose.

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The animal experiments were reviewed and approved by the animal welfare and ethics committees of the Fifth Affiliated Hospital of Sun Yat-sen University. Approval for the use of human samples was obtained from the Ethics Review Committee of the Fifth Affiliated Hospital of Sun Yat-sen University with waiver of informed consents (K170-1).

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Xiao, Y., Mei, C., Xu, D. et al. Identification of a CEACAM5 targeted nanobody for positron emission tomography imaging and near-infrared fluorescence imaging of colorectal cancer. Eur J Nucl Med Mol Imaging 50, 2305–2318 (2023). https://doi.org/10.1007/s00259-023-06183-7

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