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Preclinical evaluation of a dual-receptor targeted tracer [68Ga]Ga-HX01 in 10 different subcutaneous and orthotopic tumor models

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

Abstract

Purpose

The integrin αvβ3 and aminopeptidase N (APN/CD13) play vital roles in the tumor angiogenesis process. They are highly expressed in a variety of tumor cells and proliferating endothelial cells during angiogenesis, which have been considered as highly promising targets for tumor imaging. Arginine-glycine-aspartic (RGD) and asparagine-glycine-arginine (NGR) are two peptides specifically binding to the integrin αvβ3 and CD13, respectively. In this study, we optimized our previously developed probe and preclinically evaluated the new integrin αvβ3 and CD13 dual-targeted probe, NOTA-RGD-NGR (denoted as HX01) radiolabeled with 68Ga, in 10 different subcutaneous and orthotopic tumor models.

Methods

The specific activity and radiochemical purity of [68Ga]Ga-HX01 were identified. The dual-receptor targeting ability was confirmed by a series of blocking studies and partly muted tracers using BxPC-3 xenograft model. The dynamic imaging study and dose escalation study were explored to determine the optimal imaging time point and dosage in the BxPC-3 xenograft model. Next, we established a variety of subcutaneous and orthotopic tumor models including pancreas (BxPC-3), breast (MCF-7), gallbladder (NOZ), lung (HCC827), ovary (SK-OV-3), colorectal (HCT-8), liver (HuH-7), stomach (NUGC-4), and glioma (U87) cancers. All models underwent [68Ga]Ga-HX01 PET/CT imaging about 2 weeks post-inoculation, with a subset of them undergoing [18F]FDG PET/CT scan performed concurrently, and their results were compared. In addition, ex vivo biodistribution studies were also performed for verifying the semi-quantitative results of the non-invasive PET images.

Results

[68Ga]Ga-HX01 significantly outperformed single target probes in the BxPC-3 xenograft model. All blocking and single target groups exhibited significantly descending tumor uptake. The high tumor uptakes were found in BxPC-3, MCF-7, and NOZ subcutaneous tumors (%ID/g > 1.1), while middle uptakes were observed in HCC827, SK-OV-3, HCT-8, and HuH-7 subcutaneous tumor (%ID/g 0.7–1.0). Due to the low background, the tumor-to-muscle and tumor-to-blood ratios of [68Ga]Ga-HX01 were higher than that of [18F]FDG.

Conclusions

[68Ga]Ga-HX01, as a dual target imaging agent, exhibited superior in vivo performance in different subcutaneous and orthotopic mice models of human tumors over [18F]FDG and its respectively mono-receptor targeted agents, which warrants the future clinical translation for tumor imaging.

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Data Availability

Data and materials are available from the corresponding authors upon reasonable request.

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Acknowledgements

The authors thank Changyu Shan from Hexin (Suzhou) Pharmaceutical Technology Co., Ltd. and Jinquan Han from Wuhan-Britain China School for their help with cell culture and animal model preparing.

Funding

This work was supported by the National Natural Science Foundation of China (No. 81801738 and 82030052).

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Author notes

  1. Xiaoying Lv and Xiangming Song contributed equally.

Authors and Affiliations

  1. Department of Nuclear Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Ave 1277, Wuhan, 430022, Hubei Province, China

    Xiaoying Lv, Xiangming Song, Yu Long, Xiaoli Lan & Yongkang Gai

  2. Hubei Province Key Laboratory of Molecular Imaging, Wuhan, 430022, China

    Xiaoying Lv, Xiangming Song, Yu Long, Xiaoli Lan & Yongkang Gai

  3. Key Laboratory of Biological Targeted Therapy, The Ministry of Education, Wuhan, 430022, China

    Xiaoying Lv, Xiangming Song, Yu Long, Xiaoli Lan & Yongkang Gai

  4. Department of Radiology, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Dexing Zeng

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Contributions

Yongkang Gai and Xiaoli Lan: study design and manuscription revision. Xiaoying Lv and Xiangming Song: manuscript writing. Dexing Zeng, Yongkang Gai, Xiaoying Lv, and Xiangming Song: tracer synthesis and identification. Xiaoying Lv, Xiangming Song, and Yu Long: animal experiment, image acquisition, and interpretation.

Corresponding authors

Correspondence to Xiaoli Lan or Yongkang Gai.

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Ethical approval

All animal studies were conducted in accordance with the guidelines of the Institutional Animal Care and Use Committee of Tongji Medical College, Huazhong University of Science and Technology.

Competing interests

Dexing Zeng and Yongkang Gai hold the patent of the compound HX01. All other authors declare that they have no conflict of interests.

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Lv, X., Song, X., Long, Y. et al. Preclinical evaluation of a dual-receptor targeted tracer [68Ga]Ga-HX01 in 10 different subcutaneous and orthotopic tumor models. Eur J Nucl Med Mol Imaging 51, 54–67 (2023). https://doi.org/10.1007/s00259-023-06412-z

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