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New and effective EGFR-targeted fluorescence imaging technology for intraoperative rapid determination of lung cancer in freshly isolated tissue

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Abstract

Purpose

During lung cancer surgery, it is very important to define tumor boundaries and determine the surgical margin distance. In previous research, systemically application of fluorescent probes can help medical professionals determine the boundaries of tumors and find small tumors and metastases, thereby improving the accuracy of surgical resection. However, there are very few safe and effective probes that can be applied to clinical trials up to now, which limits the clinical application of fluorescence imaging. Here we developed a new technology that can quickly identify the tumor area in the resected lung tissue during the operation and distinguish the tumor boundary and metastatic lymph nodes.

Experimental design

For animal studies, a PDX model of lung cancer was established. The tumors, lungs, and peritumoral muscle tissues of tumor-bearing mice were surgically removed and incubated with a probe targeting epidermal growth factor receptor (EGFR) for 20 min, and then imaged by a closed-field near-infrared two-zone (NIR-II) fluorescence imaging system. For clinical samples, ten surgically removed lung tissues and 60 lymph nodes from 10 lung cancer patients undergoing radical resection were incubated with the targeting probe immediately after intraoperative resection and imaged to identify the tumor area and distinguish the tumor boundary and metastatic lymph nodes. The accuracy of fluorescence imaging was confirmed by HE staining and immunohistochemistry.

Results

The ex vivo animal imaging experiments showed a fluorescence enhancement of tumor tissue. For clinical samples, our results showed that this new technology yielded more than 85.7% sensitivity and 100% specificity in identifying the tumor area in the resected lung tissue. The average fluorescence tumor-to-background ratio was 2.5 ± 1.3. Furthermore, we also used this technique to image metastatic lymph nodes intraoperatively and showed that metastatic lymph nodes have brighter fluorescence than normal lymph nodes, as the average fluorescence tumor-to-background signal ratio was 2.7 ± 1.1. Calculations on the results of the fluorescence signal in relation to the number of metastatic lymph nodes yielded values of 77.8% for sensitivity and 92.1% for specificity. We expect this new technology to be a useful diagnostic tool for rapid intraoperative pathological detection and margin determination.

Conclusions

By using fluorescently labeled anti-human EGFR recombinant antibody scFv fragment to incubate freshly isolated tissues during surgery, the probes can quickly accumulate in lung cancer tissues, which can be used to quickly identify tumor areas in the resected lung tissues and distinguish tumor boundaries and find metastases in lymph nodes. This technology is expected to be used for rapid intraoperative pathological detection and margin determination.

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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.

Data availability

The data generated or analyzed during this study are included in the manuscript and the supplementary information files.

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Acknowledgements

This work was supported in part by the National Natural Science Foundation of China (grants no. 62027901), the Fundamental Research Funds for the Central Universities (grant no. JKF-YG-22-B005), the National Key Research and Development Program of China (under Grants 2017YFA0205200, 2016YFC0103803, and 2017YFA0700401), the National Natural Science Foundation of China (grants nos. 81227901, 81930053, 81827808, 81527805, 81971198, 81671851, 82003316, and 92059203), China Postdoctoral Science Foundation (grant nos. 2020M680301 and 2019TQ0018), Chinese Academy of Sciences Youth Innovation Promotion Association (grant no. 2018167), CAS Youth Interdisciplinary Team (JCTD-2021-08), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA16021200), Chinese Academy of Sciences Key Technology Talent Program, Project of High-Level Talents Team Introduction in Zhuhai City (Zhuhai HLHPTP201703), Beijing Natural Science Foundation (grant nos. 7214266, JQ19027), and Research and Development Fund of Peking University People’s Hospital (grant no. RS2020-05). The authors would like to acknowledge the instrumental technical support of the Multimodal Biomedical Imaging Experimental Platform, Institute of Automation, Chinese Academy of Sciences.

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Author notes

  1. Changjian Li, Jiahui Mi, and Yueqi Wang contributed equally to this work.

Authors and Affiliations

  1. School of Engineering Medicine, Beihang University, Beijing, 100191, People’s Republic of China

    Changjian Li, Zeyu Zhang & Jie Tian

  2. Key Laboratory of Big Data-Based Precision Medicine (Beihang University), Ministry of Industry and Information Technology, Beijing, 100191, People’s Republic of China

    Changjian Li, Zeyu Zhang & Jie Tian

  3. Department of Thoracic Surgery, Peking University People’s Hospital, No.11, Xi Zhi Men South Avenue, Beijing, 100190, People’s Republic of China

    Jiahui Mi & Jian Zhou

  4. CAS Key Laboratory of Molecular Imaging, Beijing Key Laboratory of Molecular Imaging, The State Key Laboratory of Management and Control for Complex Systems, Institute of Automation, Chinese Academy of Sciences, Beijing, People’s Republic of China

    Yueqi Wang, Xiaoyong Guo, Zhenhua Hu & Jie Tian

  5. Department of Gastroenterology, The Third Medical Centre, Chinese PLA General Hospital, Beijing, 100190, People’s Republic of China

    Xiaoyong Guo

  6. Zhuhai Precision Medical Center, Zhuhai People’s Hospital, Affiliated With Jinan University, Zhuhai, 519000, People’s Republic of China

    Jie Tian

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Contributions

Changjian Li synthesized the probe. Changjian Li and Jiahui Mi performed the experiments. Changjian Li and Yueqi Wang analyzed the data and wrote the manuscript. Jie Tian, Zhenhua Hu, and Jian Zhou supervised the designation of the experiments. Zeyu Zhang and Xiaoyong Guo assisted with data analysis.

Corresponding authors

Correspondence to Jian Zhou, Zhenhua Hu or Jie Tian.

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Ethics approval and consent to participate

In human subjects, the study was approved by the Institutional Review Board at the Peking University People’s Hospital (Permit Number: 2021PHB315-001). In animal subjects, all experiments were performed according to the guidelines of the Institutional Animal Care and Use Committee of Beijing Municipal Science & Technology Commission (Permit Number: 2020–0049).

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All the co-authors approved the manuscript and agreed with submission to your esteemed journal.

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The authors declare no competing interests.

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Li, C., Mi, J., Wang, Y. et al. New and effective EGFR-targeted fluorescence imaging technology for intraoperative rapid determination of lung cancer in freshly isolated tissue. Eur J Nucl Med Mol Imaging 50, 494–507 (2023). https://doi.org/10.1007/s00259-022-05975-7

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