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Application of a rapid, simple, and accurate adenovirus-based method to compare PET reporter gene/PET reporter probe systems.

ABSTRACT: PURPOSE:This study aims to use a simple, quantitative method to compare the HSV1sr39TK/(18)?F-FHBG PET reporter gene/PET reporter probe (PRG/PRP) system with PRGs derived from human nucleoside kinases. PROCEDURES:The same adenovirus vector is used to express alternative PRGs. Equal numbers of vectors are injected intravenously into mice. After PRP imaging, quantitative hepatic PET signals are normalized for transduction by measuring hepatic viral genomes. RESULTS:The same adenovirus vector was used to express equivalent amounts of HSV1sr39TK, mutant human thymidine kinase 2 (TK2-DM), and mutant human deoxycytidine kinase (dCK-A100VTM) in mouse liver. HSV1sr39TK expression was measured with (18)?F-FHBG, TK2-DM and dCK-A100VTM with (18)?F-L-FMAU. TK2-DM/(18)?F-L-FMAU and HSV1sr39TK/(18)?F-FHBG had equivalent sensitivities; dCK-A100VTM/(18)?F-L-FMAU was twice as sensitive as HSV1sr39TK/(18)?F-FHBG. CONCLUSIONS:The human PRG/PRP sensitivities are comparable and/or better than HSV1sr39TK/(18)?F-FHBG. However, for clinical use, identification of the best PRP substrate for each enzyme, characterization of probe distribution, and consequences of overexpressing nucleoside kinases must be evaluated.

SUBMITTER: Gil JS

PROVIDER: S-EPMC3833443 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature

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Application of a rapid, simple, and accurate adenovirus-based method to compare PET reporter gene/PET reporter probe systems.

Gil Jose S JS Machado Hidevaldo B HB Campbell Dean O DO McCracken Melissa M Radu Caius C Witte Owen N ON Herschman Harvey R HR

Molecular imaging and biology 20130601 3

<h4>Purpose</h4>This study aims to use a simple, quantitative method to compare the HSV1sr39TK/(18) F-FHBG PET reporter gene/PET reporter probe (PRG/PRP) system with PRGs derived from human nucleoside kinases.<h4>Procedures</h4>The same adenovirus vector is used to express alternative PRGs. Equal numbers of vectors are injected intravenously into mice. After PRP imaging, quantitative hepatic PET signals are normalized for transduction by measuring hepatic viral genomes.<h4>Results</h4>The same a ...[more]

PMID: 23054556

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Project description:PurposeRadionuclide-based reporter gene imaging has the sensitivity to monitor gene- and cell-based therapies in human subjects. Potential immunogenicity of current viral transgenes warrants development of human-based reporter systems. We compared human nucleoside kinase reporters to a panel of nucleoside analogs of FEAU, FMAU, and FIAU, including the first in vivo assessment of L-[18F]FEAU.ProceduresHuman isogenic U87 cell lines were transduced to express different human reporter genes including dCK-R104M/D133A (dCKDM), dCK-R104Q/D133N (dCKep16A), dCK-A100V/R104M/D133A (dCK3M), and TK2-N93D/L109F (TK2DM), and wild-type dCK (dCK) and herpes simplex virus type-1 (HSVTK) reporter gene as references. In vitro cell uptake assays were performed with [18F]FEAU, L-[18F]FEAU, [14C]FMAU, L-[18F]FMAU, and [124I]FIAU. Micro-positron emission tomography/X-ray computed tomography imaging of xenograft-bearing nu/nu mice was conducted with [18F]FEAU, L-[18F]FEAU, L-[18F]FMAU, and [124I]FIAU on consecutive days. A cell viability assay was also performed to assess sensitivities to gemcitabine and bromovinyldeoxyuridine (BVdU).ResultsIn vitro, dCKep16A and dCKDM with [18F]FEAU exhibited the highest sensitivity and selectivity of the human reporters, second only to HSVTK/[18F]FEAU. L-[18F]FEAU biodistribution in mice was on par with [18F]FEAU and L-[18F]FMAU. L-[18F]FMAU uptake in isogenic xenografts was highest for all human reporter genes. However, [18F]FEAU was the most selective of the short half-life reporter probes due to its minimal recognition by human dCK and relative sensitivity, whereas [124I]FIAU permitted imaging at a later time point, improving signal-to-background ratio. Of the human reporter genes, dCKep16A consistently outperformed the other tested reporters. Reporter genes of interest increased potency to the nucleoside analog prodrugs gemcitabine and BVdU.ConclusionsWe demonstrate that human nucleoside kinase reporter systems vary significantly in their sensitivity and selectivity for in vivo imaging. The sufficiently high signal-to-background ratios and enhanced suicide gene potential support clinical translation.

Dataset Information

Application of a rapid, simple, and accurate adenovirus-based method to compare PET reporter gene/PET reporter probe systems.

Publications

Application of a rapid, simple, and accurate adenovirus-based method to compare PET reporter gene/PET reporter probe systems.

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