Sensitive β-galactosidase-targeting fluorescence probe for visualizing small peritoneal metastatic tumours in vivo

Daisuke Asanuma; Masayo Sakabe; Mako Kamiya; Kyoko Yamamoto; Jun Hiratake; Mikako Ogawa; Nobuyuki Kosaka; Peter L Choyke; Tetsuo Nagano; Hisataka Kobayashi; Yasuteru Urano

doi:10.1038/ncomms7463

Sensitive β-galactosidase-targeting fluorescence probe for visualizing small peritoneal metastatic tumours in vivo

Nat Commun. 2015 Mar 13:6:6463. doi: 10.1038/ncomms7463.

Authors

Affiliations

¹ 1] Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan [2] Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
² Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
³ 1] Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan [2] PRESTO, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
⁴ Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
⁵ Institute for Chemical Research, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
⁶ Molecular Imaging Program, Center for Cancer Research, National Cancer Institute, US National Institutes of Health, Building 10, Room B3B69, Mail Stop Code 1088, 10 Center Drive, Bethesda, Maryland 20892-1088, USA.
⁷ Open Innovation Center for Drug Discovery, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
⁸ 1] Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan [2] Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan [3] CREST, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan [4] Basic Research Program, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.

Abstract

Fluorescence-guided diagnostics is one of the most promising approaches for facile detection of cancer in situ. Here we focus on β-galactosidase, which is overexpressed in primary ovarian cancers, as a molecular target for visualizing peritoneal metastases from ovarian cancers. As existing fluorescence probes are unsuitable, we have designed membrane-permeable HMRef-βGal, in which the optimized intramolecular spirocyclic function affords >1,400-fold fluorescence enhancement on activation. We confirm that HMRef-βGal sensitively detects intracellular β-galactosidase activity in several ovarian cancer lines. In vivo, this probe visualizes metastases as small as <1 mm in diameter in seven mouse models of disseminated human peritoneal ovarian cancer (SHIN3, SKOV3, OVK18, OVCAR3, OVCAR4, OVCAR5 and OVCAR8). Because of its high brightness, real-time detection of metastases with the naked eye is possible. Endoscopic fluorescence detection of metastases is also demonstrated. The results clearly indicate preclinical potential value of the probe for fluorescence-guided diagnosis of peritoneal metastases from ovarian cancers.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Disease Models, Animal
Endoscopy
Female
Fluorescent Dyes / chemistry*
Gene Expression Regulation, Neoplastic
Humans
Mice
Mice, Inbred BALB C
Microscopy, Fluorescence
Neoplasm Metastasis
Ovarian Neoplasms / diagnosis*
Ovarian Neoplasms / pathology
Peritoneal Neoplasms / diagnosis*
Peritoneal Neoplasms / secondary
beta-Galactosidase / metabolism*

Substances

Fluorescent Dyes
beta-Galactosidase