A new pyrimidine-specific reporter gene: a mutated human deoxycytidine kinase suitable for PET during treatment with acycloguanosine-based cytotoxic drugs.
In this article, we describe a series of new human-derived reporter genes based on human deoxycytidine kinase (dCK) suitable for clinical PET.


METHODS
Native dCK and its mutant reporter genes were tested in vitro and in vivo for their phosphorylation of pyrimidine- and acycloguanosine-based radiotracers including 2'-deoxy-2'-fluoroarabinofuranosylcytosine, 2'-fluoro-2'-deoxyarabinofuranosyl-5-ethyluracil (FEAU), penciclovir, and 9-[4-fluoro-3-(hydroxymethyl)butyl]guanine (FHBG) and clinically applied antiviral and anticancer drugs.


RESULTS
Cells transduced with dCK mutant reporter genes showed high in vitro and in vivo uptake of pyrimidine-based radiopharmaceuticals ((18)F-FEAU) comparable to that of herpes simplex virus type-1 thymidine kinase (HSV1-tk)-transduced cells. These mutants did not phosphorylate acycloguanosine-based radiotracers ((18)F-FHBG) or antiviral drugs (ganciclovir). Furthermore, the mutants displayed suicidal activation of clinically used pyrimidine-based prodrugs (cytarabine, gemcitabine).


CONCLUSIONS
The mutants of human dCK can be used as pyrimidine-specific PET reporter genes for imaging with (18)F-FEAU during treatment with acycloguanosine-based antiviral drugs. Additionally, the prosuicidal activity of these reporters with pyrimidine-based analogs will allow for the safe elimination of transduced cells.
Likar
Yury
Y
Department of Radiology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Zurita
Juan
J
Dobrenkov
Konstantin
K
Shenker
Larissa
L
Cai
Shangde
S
Neschadim
Anton
A
Medin
Jeffrey A
JA
Sadelain
Michel
M
Hricak
Hedvig
H
Ponomarev
Vladimir
V
D000212:Acyclovir / Q000737:chemistry* / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D001086:Arabinofuranosyluracil / Q000031:analogs & derivatives* / Q000378:metabolism; D045744:Cell Line, Tumor; D003842:Deoxycytidine Kinase / Q000235:genetics* / Q000378:metabolism; D005462:Fluorine Radioisotopes; D017930:Genes, Reporter / Q000235:genetics*; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D051379:Mice; D009154:Mutation; D041681:NIH 3T3 Cells; D010766:Phosphorylation; D049268:Positron-Emission Tomography; D011355:Prodrugs / Q000494:pharmacology; D011849:Radioactive Tracers; D013379:Substrate Specificity; D014057:Tomography, X-Ray Computed; D014161:Transduction, Genetic
D000970:Antineoplastic Agents; D005462:Fluorine Radioisotopes; D011355:Prodrugs; D011849:Radioactive Tracers; D001086:Arabinofuranosyluracil; C054103:2'-fluoro-5-ethylarabinosyluracil; D003842:Deoxycytidine Kinase; D000212:Acyclovir
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