dc.contributor.author |
Mowday, Alexandra Marie |
|
dc.contributor.author |
Copp, Janine Naomi |
|
dc.contributor.author |
Syddall, Sophie Philippa |
|
dc.contributor.author |
Dubois, Ludwig Jerome |
|
dc.contributor.author |
Wang, Jingli |
|
dc.contributor.author |
Lieuwes, Natasja Gabi |
|
dc.contributor.author |
Biemans, Rianne |
|
dc.contributor.author |
Ashoorzadeh, Amir |
|
dc.contributor.author |
Abbattista, Maria Rosaria |
|
dc.contributor.author |
Williams, Elsie May |
|
dc.contributor.author |
Guise, Christopher Paul |
|
dc.contributor.author |
Lambin, Philippe |
|
dc.contributor.author |
Ackerley, David Francis |
|
dc.contributor.author |
Smaill, Jeff Bruce |
|
dc.contributor.author |
Theys, Jan |
|
dc.contributor.author |
Patterson, Adam Vorn |
|
dc.coverage.spatial |
Australia |
|
dc.date.accessioned |
2023-02-22T22:48:36Z |
|
dc.date.available |
2023-02-22T22:48:36Z |
|
dc.date.issued |
2020-01 |
|
dc.identifier.citation |
(2020). Theranostics, 10(23), 10548-10562. |
|
dc.identifier.issn |
1838-7640 |
|
dc.identifier.uri |
https://hdl.handle.net/2292/62987 |
|
dc.description.abstract |
The use of reporter genes to non-invasively image molecular processes inside cells has significant translational potential, particularly in the context of systemically administered gene therapy vectors and adoptively administered cells such as immune or stem cell based therapies. Bacterial nitroreductase enzymes possess ideal properties for reporter gene imaging applications, being of non-human origin and possessing the ability to metabolize a range of clinically relevant nitro(hetero)cyclic substrates. <b>Methods:</b> A library of eleven <i>Escherichia coli</i> nitroreductase candidates were screened for the ability to efficiently metabolize 2-nitroimidazole based positron emission tomography (PET) probes originally developed as radiotracers for hypoxic cell imaging. Several complementary methods were utilized to detect formation of cell-entrapped metabolites, including various <i>in vitro</i> and <i>in vivo</i> models to establish the capacity of the 2-nitroimidazole PET agent EF5 to quantify expression of a nitroreductase candidate. Proof-of-principle PET imaging studies were successfully conducted using <sup>18</sup>F-HX4. <b>Results:</b> Recombinant enzyme kinetics, bacterial SOS reporter assays, anti-proliferative assays and flow cytometry approaches collectively identified the major oxygen-insensitive nitroreductase NfsA from <i>E. coli</i> (NfsA_Ec) as the most promising nitroreductase reporter gene. Cells expressing NfsA_Ec were demonstrably labelled with the imaging agent EF5 in a manner that was quantitatively superior to hypoxia, in monolayers (2D), multicellular layers (3D), and in human tumor xenograft models. EF5 retention correlated with NfsA_Ec positive cell density over a range of EF5 concentrations in 3D <i>in vitro</i> models and in xenografts <i>in vivo</i> and was predictive of <i>in vivo</i> anti-tumor activity of the cytotoxic prodrug PR-104. Following PET imaging with <sup>18</sup>F-HX4, a significantly higher tumor-to-blood ratio was observed in two xenograft models for NfsA_Ec expressing tumors compared to the parental tumors thereof, providing verification of this reporter gene imaging approach. <b>Conclusion:</b> This study establishes that the bacterial nitroreductase NfsA_Ec can be utilized as an imaging capable reporter gene, with the ability to metabolize and trap 2-nitroimidazole PET imaging agents for non-invasive imaging of gene expression. |
|
dc.format.medium |
Electronic-eCollection |
|
dc.language |
eng |
|
dc.publisher |
Ivyspring International Publisher |
|
dc.relation.ispartofseries |
Theranostics |
|
dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. |
|
dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
|
dc.rights.uri |
https://creativecommons.org/licenses/by/4.0/ |
|
dc.subject |
HCT116 Cells |
|
dc.subject |
Animals |
|
dc.subject |
Humans |
|
dc.subject |
Mice |
|
dc.subject |
Neoplasms |
|
dc.subject |
Hydrocarbons, Fluorinated |
|
dc.subject |
Nitrogen Mustard Compounds |
|
dc.subject |
Etanidazole |
|
dc.subject |
Triazoles |
|
dc.subject |
Imidazoles |
|
dc.subject |
Nitroreductases |
|
dc.subject |
Escherichia coli Proteins |
|
dc.subject |
Recombinant Proteins |
|
dc.subject |
Antineoplastic Agents, Alkylating |
|
dc.subject |
Indicators and Reagents |
|
dc.subject |
Radiopharmaceuticals |
|
dc.subject |
Positron-Emission Tomography |
|
dc.subject |
Xenograft Model Antitumor Assays |
|
dc.subject |
Drug Resistance, Neoplasm |
|
dc.subject |
Genes, Reporter |
|
dc.subject |
Genetic Vectors |
|
dc.subject |
Molecular Imaging |
|
dc.subject |
Genetic Therapy |
|
dc.subject |
Precision Medicine |
|
dc.subject |
Tumor Hypoxia |
|
dc.subject |
Proof of Concept Study |
|
dc.subject |
PET imaging |
|
dc.subject |
drug repurposing |
|
dc.subject |
gene therapy |
|
dc.subject |
nitroreductase |
|
dc.subject |
reporter gene imaging |
|
dc.subject |
Bioengineering |
|
dc.subject |
Genetics |
|
dc.subject |
Cancer |
|
dc.subject |
Biomedical Imaging |
|
dc.subject |
Biotechnology |
|
dc.subject |
5.2 Cellular and gene therapies |
|
dc.subject |
5 Development of treatments and therapeutic interventions |
|
dc.subject |
Science & Technology |
|
dc.subject |
Life Sciences & Biomedicine |
|
dc.subject |
Medicine, Research & Experimental |
|
dc.subject |
Research & Experimental Medicine |
|
dc.subject |
POSITRON-EMISSION-TOMOGRAPHY |
|
dc.subject |
BIOREDUCTIVE PRODRUG PR-104A |
|
dc.subject |
PHASE-I |
|
dc.subject |
ANTITUMOR-ACTIVITY |
|
dc.subject |
EXPRESSION |
|
dc.subject |
ACTIVATE |
|
dc.subject |
OXIDOREDUCTASE |
|
dc.subject |
BIOMARKER |
|
dc.subject |
ABLATION |
|
dc.subject |
1112 Oncology and Carcinogenesis |
|
dc.title |
<i>E. coli</i> nitroreductase NfsA is a reporter gene for non-invasive PET imaging in cancer gene therapy applications. |
|
dc.type |
Journal Article |
|
dc.identifier.doi |
10.7150/thno.46826 |
|
pubs.issue |
23 |
|
pubs.begin-page |
10548 |
|
pubs.volume |
10 |
|
dc.date.updated |
2023-01-10T05:32:03Z |
|
dc.rights.holder |
Copyright: The authors |
en |
dc.identifier.pmid |
32929365 (pubmed) |
|
pubs.author-url |
https://www.ncbi.nlm.nih.gov/pubmed/32929365 |
|
pubs.end-page |
10562 |
|
pubs.publication-status |
Published |
|
dc.rights.accessrights |
http://purl.org/eprint/accessRights/OpenAccess |
en |
pubs.subtype |
Research Support, Non-U.S. Gov't |
|
pubs.subtype |
research-article |
|
pubs.subtype |
Journal Article |
|
pubs.elements-id |
816853 |
|
pubs.org-id |
Medical and Health Sciences |
|
pubs.org-id |
Science |
|
pubs.org-id |
Science Research |
|
pubs.org-id |
Medical Sciences |
|
pubs.org-id |
Auckland Cancer Research |
|
pubs.org-id |
Maurice Wilkins Centre (2010-2014) |
|
dc.identifier.eissn |
1838-7640 |
|
dc.identifier.pii |
thnov10p10548 |
|
pubs.record-created-at-source-date |
2023-01-10 |
|
pubs.online-publication-date |
2020-08-21 |
|