Research Papers:

A topically-sprayable, activatable fluorescent and retaining probe, SPiDER-βGal for detecting cancer: Advantages of anchoring to cellular proteins after activation

Yuko Nakamura, Ai Mochida, Tadanobu Nagaya, Shuhei Okuyama, Fusa Ogata, Peter L. Choyke and Hisataka Kobayashi _

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Oncotarget. 2017; 8:39512-39521. https://doi.org/10.18632/oncotarget.17080

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Yuko Nakamura1, Ai Mochida1, Tadanobu Nagaya1, Shuhei Okuyama1, Fusa Ogata1, Peter L. Choyke1 and Hisataka Kobayashi1

1Molecular Imaging Program, Center for Cancer Research, National Cancer Institute, United States National Institutes of Health, Bethesda, Maryland 20892-1088, USA

Correspondence to:

Hisataka Kobayashi, email: kobayash@mail.nih.gov

Keywords: activatable probe, β-galactosidase, ovarian cancer, preservation of fluorescence, target-to-background ratios

Received: January 17, 2017     Accepted: March 22, 2017     Published: April 13, 2017


SPiDER-βGal is a newly-developed probe that is activated by β-galactosidase and is then retained within cells by anchoring to intracellular proteins. Previous work has focused on gGlu-HMRG, a probe activated by γ-glutamyltranspeptidase, which demonstrated high sensitivity for the detection of peritoneal ovarian cancer metastases in an animal model. However, its fluorescence, after activation by γ-glutamyltranspeptidase, rapidly declines over time, limiting the actual imaging window and the ability to define the border of lesions. The purpose of this study is to compare the fluorescence signal kinetics of SPiDER-βGal with that of gGlu-HMRG using ovarian cancer cell lines in vitro and ex vivo tissue imaging. In vitro removal of gGlu-HMRG resulted in a rapid decrease of fluorescence intensity followed by a more gradual decrease up to 60 min while there was a gradual increase in fluorescence up to 60 min after removal of SPiDER-βGal. This is most likely due to internalization and retention of the dye within cells. This was also confirmed ex vivo tissue imaging using a red fluorescence protein (RFP)-labeled tumor model in which the intensity of fluorescence increased gradually after activation of SPiDER-βGal. Additionally, SPiDER-βGal resulted in intense enhancement within the tumor due to the high target-to-background ratio, which extended up to 60 min after activation. In contrast, gGlu-HMRG fluorescence resulted in decreasing fluorescence over time in extracted tumors. Thus, SPiDER-βGal has the advantages of higher signal with more signal retention, resulting in improved contrast of the tumor margin and suggesting it may be an alternative to existing activatable probes.

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