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Preclinical evaluation of a novel EGFR&c-Met bispecific near infrared probe for visualization of esophageal cancer and metastatic lymph nodes

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

Abstract

Purpose

This study aimed to establish a near infrared fluorescent (NIRF) probe based on an EGFR&c-Met bispecific antibody for visualization of esophageal cancer (EC) and metastatic lymph nodes (mLNs).

Methods

EGFR and c-Met expression were assessed by immunohistochemistry. EGFR&c-Met bispecific antibody EMB01 was labeled with IRDye800cw. The binding of EMB01-IR800 was assessed by enzyme linked immunosorbent assay, flow cytometry, and immunofluorescence. Subcutaneous tumors, orthotopic tumors, and patient-derived xenograft (PDX) were established for in vivo fluorescent imaging. PDX models using lymph nodes with or without metastasis were constructed to assess the performance of EMB01-IR800 in differential diagnosis of lymph nodes.

Results

The prevalence of overexpressing EGFR or c-Met was significantly higher than single marker either in EC or corresponding mLNs. The bispecific probe EMB01-IR800 was successfully synthesized, with strong binding affinity. EMB01-IR800 showed strong cellular binding to both Kyse30 (EGFR overexpressing) and OE33 (c-Met overexpressing) cells. In vivo fluorescent imaging showed prominent EMB01-IR800 uptake in either Kyse30 or OE33 subcutaneous tumors. Likewise, EMB01-IR800 exhibited superior tumor enrichment in both thoracic orthotopic esophageal squamous cell carcinoma and abdominal orthotopic esophageal adenocarcinoma models. Moreover, EMB01-IR800 produced significantly higher fluorescence in patient-derived mLNs than in benign lymph nodes.

Conclusion

This study demonstrated the complementary overexpression of EGFR and c-Met in EC. Compared to single-target probes, the EGFR&c-Met bispecific NIRF probe can efficiently depict heterogeneous esophageal tumors and mLNs, which greatly increased the sensitivity of tumor and mLN identification.

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Data availability

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

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Funding

This work was supported by the National Natural Science Foundation of China (92259204) and the Natural Science Foundation of Guangdong Province (2022A1515012509).

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Author notes

  1. Mingzhu Liang, Lizhu Wang, and Yitai Xiao contributed equally.

Authors and Affiliations

  1. Department of Radiology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, 519000, China

    Mingzhu Liang, Lizhu Wang & Yaqin Zhang

  2. Center for Interventional Medicine, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, 519000, China

    Mingzhu Liang, Lizhu Wang, Yaqin Zhang & Hong Shan

  3. Department of Nuclear Medicine, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, 519000, China

    Yitai Xiao, Meilin Yang, Chaoming Mei & Dan Li

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

    Yitai Xiao, Meilin Yang, Chaoming Mei, Hong Shan & Dan Li

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  1. Mingzhu Liang
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Contributions

Dan Li, Yaqin Zhang, and Hong Shan contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Mingzhu Liang, Lizhu Wang, Yitai Xiao, Meilin Yang, and Chaoming Mei. The manuscript was written by Mingzhu Liang and Yitai Xiao. All the authors revised previous versions of the manuscript. All the authors read and approved the final manuscript.

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Correspondence to Yaqin Zhang, Hong Shan or Dan Li.

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The use of pathologic sections had been approved by ethical review board of the Fifth Affiliated Hospital of Sun Yat-sen University with waiving of informed consents. The animal experiments have been approved by the animal welfare committee of the Fifth Affiliated Hospital of Sun Yat-sen University (00094).

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Liang, M., Wang, L., Xiao, Y. et al. Preclinical evaluation of a novel EGFR&c-Met bispecific near infrared probe for visualization of esophageal cancer and metastatic lymph nodes. Eur J Nucl Med Mol Imaging 50, 2787–2801 (2023). https://doi.org/10.1007/s00259-023-06250-z

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