Evaluation of an Integrin αvβ3 and Aminopeptidase N Dual-Receptor Targeting Tracer for Breast Cancer Imaging

Yongkang Gai, Yaqun Jiang, Yu Long, Lingyi Sun, Qingyao Liu, Chunxia Qin, Yongxue Zhang, Dexing Zeng, Xiaoli Lan

Research output: Contribution to journalArticle

Abstract

Integrin αvβ3 and aminopeptidase N (APN, also known as CD13) are two important targets involved in the regulation of angiogenesis, tumor proliferation, invasion, and metastasis. In this study, we developed a heterodimeric tracer consisting of arginine-glycine-aspartic (RGD) and asparagine-glycine-arginine (NGR) peptides targeting αvβ3 and CD13, respectively, for PET imaging of breast cancer. The NGR peptide was first modified with N3-NOtB2 and then conjugated to BCN-PEG4-c(RGDyK) via copper-free click chemistry. The resulting precursor was purified and radiolabeled with gallium-68. Small-animal PET/CT imaging and post-imaging biodistribution studies were performed in mice bearing human breast cancer MCF-7, MDA-MB-231, MDA-MB-468, and MX-1 xenografts and pulmonary metastases models. The expression levels of αvβ3 and CD13 in tumors were checked via immunochemical staining. The heterodimeric tracer was successfully synthesized and radiolabeled with gallium-68 at a molar activity of 45-100 MBq/nmol at the end of synthesis. It demonstrated high in vitro and in vivo stability. In static PET/CT imaging studies, the MCF-7 tumor could be clearly visualized and exhibited higher uptake at 30 min post injection of 68Ga-NGR-RGD than that of either 68Ga-RGD or 68Ga-NGR alone. High specificity was shown in blocking studies using Arg-Gly-Asp (RGD) and Asp-Gly-Arg (NGR) peptides. The MCF-7 tumor exhibited the highest uptake of 68Ga-NGR-RGD followed by MDA-MB-231, MDA-MB-468, and MX-1 tumors. This was consistent with their expression levels of CD13 and αvβ3 as confirmed by western blot and immunohistochemical staining. Metastatic lesions in the lungs were clearly detectable on 68Ga-NGR-RGD PET/CT imaging in mouse models of pulmonary metastases. 68Ga-NGR-RGD, a CD13 and αvβ3 dual-receptor targeting tracer, showed higher binding avidities, targeting efficiency, and longer tumor retention time compared with monomeric 68Ga-NGR and 68Ga-RGD. Its promising in vivo performance makes it an ideal candidate for future clinical translation.

Original languageEnglish (US)
JournalMolecular Pharmaceutics
DOIs
StateAccepted/In press - Jan 1 2020

Fingerprint

CD13 Antigens
NGR peptide
Integrins
Breast Neoplasms
Neoplasms
Gallium
Neoplasm Metastasis
Lung
Glycine
Arginine
Click Chemistry
Staining and Labeling
Asparagine
Heterografts
Copper
Western Blotting
Injections

Keywords

  • angiogenesis
  • breast cancer
  • CD13
  • dual-receptor targeted
  • integrin αβ

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmaceutical Science
  • Drug Discovery

Cite this

Evaluation of an Integrin αvβ3 and Aminopeptidase N Dual-Receptor Targeting Tracer for Breast Cancer Imaging. / Gai, Yongkang; Jiang, Yaqun; Long, Yu; Sun, Lingyi; Liu, Qingyao; Qin, Chunxia; Zhang, Yongxue; Zeng, Dexing; Lan, Xiaoli.

In: Molecular Pharmaceutics, 01.01.2020.

Research output: Contribution to journalArticle

Gai, Yongkang ; Jiang, Yaqun ; Long, Yu ; Sun, Lingyi ; Liu, Qingyao ; Qin, Chunxia ; Zhang, Yongxue ; Zeng, Dexing ; Lan, Xiaoli. / Evaluation of an Integrin αvβ3 and Aminopeptidase N Dual-Receptor Targeting Tracer for Breast Cancer Imaging. In: Molecular Pharmaceutics. 2020.
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AU - Gai, Yongkang

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AU - Long, Yu

AU - Sun, Lingyi

AU - Liu, Qingyao

AU - Qin, Chunxia

AU - Zhang, Yongxue

AU - Zeng, Dexing

AU - Lan, Xiaoli

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