In-vivo PET imaging of inducible D2R reporter transgene expression using [11C]FLB 457 as reporter probe in living rats

BackgroundIncreasing interest is being shown in a variety of methods for the in-vivo monitoring of gene expression. Of these, the reporter assay using positron emission tomography (PET) has been studied most extensively. MethodsWe evaluated tetracycline-induced gene expression using a PET reporter method employing the dopamine type 2 receptor (D2R) gene as a reporter gene and [11C]FLB 457 as a reporter probe. We constructed a plasmid containing the D2R gene, whose expression was under the control of the tetracycline-responsive element, and transfected it into HeLa-Tet-On cells. D2R messenger RNA (mRNA) expression was measured by reverse transcription-polymerase chain reaction (RT-PCR) and D2R binding in the cultured cells was measured by a binding assay using methoxy-[3H]raclopride as a ligand. The tetracycline analogue, doxycycline, was used to regulate D2R expression. ResultsDoxycycline dose- and exposure time-dependent D2R transgene expression was observed in the mRNA measurements and receptor binding in the cells. The stably transfected cells were inoculated into nude rats and D2R expression in xenograft tumours was monitored by in-vivo receptor binding using PET. Doxycycline-dependent D2R expression was also observed in this in-vivo system. The correlation between the magnitude of the [11C]FLB 457 PET signal and the D2R-expressing cell fraction in the tumours showed the usefulness of the D2R–FLB 457 reporter gene–reporter probe system with PET for the quantitative evaluation of inducible in-vivo gene expression. ConclusionThe D2R–FLB 457 reporter gene–reporter probe system should be considered as a useful technique for measuring inducible in-vivo gene expression.