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GPC3-targeted immunoPET imaging of hepatocellular carcinomas

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

Abstract

Purpose

Early detection of hepatocellular carcinoma (HCC) remains a clinical challenge. Glypican 3 (GPC3) is a proteoglycan highly specific for HCC and is a potential diagnostic and therapeutic target for HCC. This work aims to develop GPC3-targeted immuno-positron emission tomography (immunoPET) imaging strategies and to assess the diagnostic values in preclinical HCC models.

Methods

Flow cytometry was used to screen GPC3-positive HCC cell lines. The expression of GPC3 in HCCs was detected by immunohistochemistry on tissue microarray. A novel GPC3-specific single domain antibody (sdAb) was produced and labeled with gallium-68 (68Ga, T1/2 = 1.1 h) and fluorine-18 (18F, T1/2 = 1.8 h) to develop radiotracers with different half-lives. The diagnostic efficacies of the developed probes (i.e., [68Ga]Ga-NOTA-G2, [18F]F-G2, and [68Ga]Ga-NOTA-ABDG2) were interrogated in preclinical HCC models bearing varying GPC3 levels.

Results

GPC3 was strongly expressed on HCC cell lines and patients with poorly differentiated HCC. [68Ga]Ga-NOTA-G2 immunoPET imaging specifically delineated the subcutaneous HCC lesions, outperforming the traditional 18F-fluorodeoxyglucose PET and the nonspecific [68Ga]Ga-NOTA-NbGFP immunoPET. ImmunoPET imaging with [18F]F-G2 also efficiently diagnosed the tumors with clarity. Moreover, the fusion of G2 to an albumin-binding domain (ABD) significantly increased the tumor uptake and decreased kidney accumulation of the radiotracer when compared to [68Ga]Ga-NOTA-G2.

Conclusions

In the work, we successfully developed sdAb-derived GPC3-targeted immunoPET imaging strategies and characterized the superior diagnostic accuracies in preclinical HCC models. Furthermore, we synthesized a fusion protein ABDG2 with improved targeting and pharmacokinetic properties, serving as a promising candidate for developing radioimmunotherapy agents.

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Acknowledgements

We thank colleagues at our department for the helpful discussions.

Funding

This research was funded in part by the National Key Research and Development Program of China (Grant No. 2020YFA0909000 and 2021YFA0910000), the National Natural Science Foundation of China (Grant No. 82001878 and 82171972), the Shanghai Rising-Star Program (Grant No. 20QA1406100), and the Interdisciplinary Program of Shanghai Jiao Tong University (Grant No.YG2019QNA27).

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Author notes

  1. Shuxian An and Di Zhang contributed equally to this work.

Authors and Affiliations

  1. Department of Nuclear Medicine, School of Medicine, Renji Hospital, Shanghai Jiao Tong University, 1630 Dongfang Rd, Shanghai, 200127, China

    Shuxian An, Di Zhang, You Zhang, Cheng Wang, Weijun Wei, Gang Huang & Jianjun Liu

  2. Department of Nuclear Medicine, Nanjing First Hospital, Nanjing Medical University, 68 Changle Rd, Nanjing, 210006, China

    Liang Shi

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  1. Shuxian An
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Contributions

W. Wei, S. An, G. Huang, and J. Liu together designed the project. W. Wei, S. An, and G. Huang obtained the funds supporting the work. S. An and D. Zhang performed the majority of the experiments, under the supervision of W. Wei, G. Huang, and J. Liu, and with the help of Y. Zhang and C. Wang. S. An and L. Shi together analyzed the tissue microarray data and clinical data. S. An drafted the manuscript and the three senior authors revised and finalized the manuscript.

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Correspondence to Weijun Wei, Gang Huang or Jianjun Liu.

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Conflict of interest

W. Wei, S. An, and J. Liu are co-inventors on a provisional patent application encompassing the technologies developed in the work. No other competing interests exist.

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This article is part of the Topical Collection on Preclinical Imaging.

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An, S., Zhang, D., Zhang, Y. et al. GPC3-targeted immunoPET imaging of hepatocellular carcinomas. Eur J Nucl Med Mol Imaging 49, 2682–2692 (2022). https://doi.org/10.1007/s00259-022-05723-x

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