Non-invasive optical imaging of cathepsin B with activatable fluorogenic nanoprobes in various metastatic models

Abstract

An increasing number of treatments of metastases rely on diagnostics and imaging these days. The facts that the activity of cathepsin B (CB) is markedly linked to the metastatic process and that CB is found highly expressed in the pericellular regions in this process make CB an attractive target for diagnosing metastases. We have developed a CB-sensitive nanoprobe (CB-CNP) consisting of self-quenched CB-sensitive fluorogenic peptide probes conjugated onto the surface of tumor-targeting glycol chitosan nanoparticles (CNPs). The freshly prepared CB-CNP formed a spherical nanoparticle structure (280 nm in diameter) and the fluorescence intensity of CB-CNP was strongly quenched in physiological condition. However, self-quenched CB-CNP boosted strong fluorescence signals in the presence of CB, not of cathepsin I. or cathepsin D, due to the CB-specific cleavage of self-quenched peptide probes. Importantly, the intravenously injected CB-CNP demonstrated the potential to discriminate metastases in vivo in three metastatic mouse models, including 4T1-luc2 liver metastases, RFP-B16F10 lung metastases and HT1080 peritoneal metastases. Indeed, Western blot analysis confirmed that the CB expression of metastases had increased compared to normal organ in these metastatic mouse models. CB-CNPs may be useful for depicting metastases through non-invasive CB molecular imaging. (C) 2013 Elsevier Ltd. All rights reserved.