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A rationally designed nuclei-targeting FAPI 04-based molecular probe with enhanced tumor uptake for PET/CT and fluorescence imaging

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

Abstract

Purpose

Fibroblast activation protein inhibitor (FAPI) -based probes have been widely studied in the diagnosis of various malignant tumors with positron emission tomography/computed tomography (PET/CT). However, current imaging studies of FAPI-based probes face challenges in rapid clearance rate and potential false-negative results. Furthermore, FAPI has been rarely explored in optical imaging. Considering this, further modifications are imperative to improve the properties of FAPI-based probes to address existing limitations and broaden their application scenarios. In this study, we rationally introduced methylene blue (MB) to FAPIs, thereby imparting nuclei-targeting and fluorescence imaging capabilities to the probes. Furthermore, we evaluated the added value of FAPI-based fluorescence imaging to traditional PET/CT, exploring the potential application of FAPI-based probes in intraoperative fluorescence imaging.

Methods

A new FAPI-based probe, namely NOTA-FAPI-MB, was designed for both PET/CT and fluorescence imaging by conjugation of MB. The targeting efficacy of the probe was evaluated on fibroblast activation protein (FAP)-transfected cell line and human primary cancer-associated fibroblasts (CAFs). Subsequently, PET/CT and fluorescence imaging were conducted on tumor-bearing mice. The tumor detection and boundary delineation were assessed by fluorescence imaging of tissues from hepatocellular carcinoma (HCC) patients.

Results

NOTA-FAPI-MB demonstrated exceptional targeting ability towards FAP-transfected cells and CAFs in comparison to NOTA-FAPI. This benefit arises from the cationic methylene blue (MB) affinity for anionic nucleic acids. PET/CT imaging of tumor-bearing mice revealed significantly higher tumor uptake of [18F]F-NOTA-FAPI-MB (standard uptake value of 2.20 ± 0.31) compared to [18F]F-FDG (standard uptake value of 1.66 ± 0.14). In vivo fluorescence imaging indicated prolonged retention at the tumor site, with retention lasting up to 24 h. In addition, the fluorescent probes enabled more precise lesion detection and tumor margin delineation than clinically used indocyanine green (ICG), achieving a 100.0% (6/6) tumor-positive rate for NOTA-FAPI-MB while 33.3% (2/6) for ICG. These findings highlighted the potential of NOTA-FAPI-MB in guiding intraoperative surgical procedures.

Conclusions

The NOTA-FAPI-MB was successfully synthesized, in which FAPI and MB simultaneously contributed to the targeting effect. Notably, the nuclear delivery mechanism of the probes improved intracellular retention time and targeting efficacy, broadening the imaging time window for fluorescence imaging. In vivo PET/CT demonstrated favorable performance of NOTA-FAPI-MB compared to [18F]F-FDG. This study highlights the significance of fluorescence imaging as an adjunct technique to PET/CT. Furthermore, the encouraging results obtained from the imaging of human HCC tissues hold promise for the potential application of NOTA-FAPI-MB in intraoperative fluorescent surgery guidance within clinical settings.

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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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Funding

This work was supported by the National Natural Science Foundation of China (82372010 & 22077061), National Key R&D Program of China (2023YFF0715303), Guangdong Provincial People’s Hospital Supporting Fund for Talent Program (KJ012020638) and Guangdong Provincial Key Laboratory of Tumor Interventional Diagnosis and Treatment (2021B1212040004).

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Authors and Affiliations

  1. Medical Research Institute, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, China

    Xingyang Zhao, Jiali Chen, Zirong Li & Liming Nie

  2. PET Center, Department of Nuclear Medicine, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, China

    Guojin Zhang & Chuangyan Zhai

  3. School of Forensic Medicine, Southern Medical University, Guangzhou, 510515, China

    Chuangyan Zhai

  4. Department of Radiation Oncology, Zhuhai People’s Hospital, Zhuhai Hospital Affiliated with Jinan University, Zhuhai, 519000, China

    Yusheng Shi

  5. Research and Development Center, Guangdong Huixuan Pharmaceutical Technology Co., Ltd, Guangzhou, 510765, China

    Guiting Li

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Contributions

Liming Nie, Chuangyan Zhai were responsible for the conception and design of the study, critical revision of the manuscript for important intellectual content and final approval of the version to be published. Xingyang Zhao, Guojin Zhang, Zirong Li were responsible for the the acquisition, analysis and interpretation of the data, and the drafting of the manuscript. Jiali Chen assisted the analysis of the data and revision of the manuscripts. Yusheng Shi and Guiting Li assisted the analysis of the data.

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Correspondence to Chuangyan Zhai or Liming Nie.

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The authors declare no competing interests.

Ethical approval

For human tissues, the study was approved by the ethics committee of Guangdong provincial people′s hospital (S2023-1055-01). For animal subjects, all experimental protocols were approved by the ethics committee of Guangdong provincial people′s hospital (2021006).

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Informed consent was obtained from all individual participants included in the study.

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Zhao, X., Zhang, G., Chen, J. et al. A rationally designed nuclei-targeting FAPI 04-based molecular probe with enhanced tumor uptake for PET/CT and fluorescence imaging. Eur J Nucl Med Mol Imaging 51, 1593–1604 (2024). https://doi.org/10.1007/s00259-024-06691-0

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