Development of a Bioluminescent Nitroreductase Probe for Preclinical Imaging

Anzhelika G Vorobyeva; Michael Stanton; Aurélien Godinat; Kjetil B Lund; Grigory G Karateev; Kevin P Francis; Elizabeth Allen; Juri G Gelovani; Emmet McCormack; Mark Tangney; Elena A Dubikovskaya

doi:10.1371/journal.pone.0131037

Development of a Bioluminescent Nitroreductase Probe for Preclinical Imaging

PLoS One. 2015 Jun 25;10(6):e0131037. doi: 10.1371/journal.pone.0131037. eCollection 2015.

Affiliations

¹ School of Basic Sciences, Institute of Chemical Sciences and Engineering, Swiss Federal Institute of Technology of Lausanne, Lausanne, Switzerland.
² Cork Cancer Research Centre, University College Cork, Cork, Ireland.
³ Department of Clinical Science, University of Bergen, Bergen, Norway; Department of Internal Medicine, Hematology Section, Haukeland University Hospital, Bergen, Norway.
⁴ PerkinElmer, Hopkinton, Massachusetts, United States of America.
⁵ School of Life Sciences, Swiss Institute for Experimental Cancer Research (ISREC), Swiss Federal Institute of Technology of Lausanne, Lausanne, Switzerland.
⁶ Department of Biomedical Engineering, College of Engineering and School of Medicine, Wayne State University, Detroit, Michigan, United States of America.

Abstract

Bacterial nitroreductases (NTRs) have been widely utilized in the development of novel antibiotics, degradation of pollutants, and gene-directed enzyme prodrug therapy (GDEPT) of cancer that reached clinical trials. In case of GDEPT, since NTR is not naturally present in mammalian cells, the prodrug is activated selectively in NTR-transformed cancer cells, allowing high efficiency treatment of tumors. Currently, no bioluminescent probes exist for sensitive, non-invasive imaging of NTR expression. We therefore developed a "NTR caged luciferin" (NCL) probe that is selectively reduced by NTR, producing light proportional to the NTR activity. Here we report successful application of this probe for imaging of NTR in vitro, in bacteria and cancer cells, as well as in vivo in mouse models of bacterial infection and NTR-expressing tumor xenografts. This novel tool should significantly accelerate the development of cancer therapy approaches based on GDEPT and other fields where NTR expression is important.

Publication types

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

MeSH terms

Animals
Cell Line
Diagnostic Imaging / methods*
Female
Genetic Therapy / methods
Humans
Luminescent Measurements / methods*
Mice
Mice, Inbred BALB C
Neoplasms / diagnosis*
Nitroreductases / metabolism*
Prodrugs / pharmacology

Substances

Prodrugs
Nitroreductases

Grants and funding

This work was supported by Intrace Medical, SA (Switzerland), Industrial grant 8421 to EAD, Bergen Research Foundation, Norwegian Cancer Society and the Western regional health board of Norway to EM, Science Foundation Ireland/Enterprise Ireland (12/TIDA/B2437), the Irish Health Research Board PhD Scholars Programme in Cancer Biology and grant HRA_POR/2010/138 and the Irish Cancer Society (PCI12TAN), the European Commission Seventh Framework Programme (PIAP-GA-2013-612219-VIP) to MT. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. PerkinElmer provided support in the form of a salary for author KPF, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific role of this author is articulated in the 'author contributions' section.