TY - JOUR T1 - Cellular uptake of [125I]-FIAU in cultured neonate rat myocardial cells after transfer HSV1-tk gene JF - Journal of Nuclear Medicine JO - J Nucl Med SP - 226P LP - 226P VL - 48 IS - supplement 2 AU - Xiao-li Lan AU - Rui-hua Wang AU - Yong He AU - Guo-xiang Cao AU - Zai-rong Gao AU - Rui An AU - Yong-xue Zhang Y1 - 2007/05/01 UR - http://jnm.snmjournals.org/content/48/supplement_2/226P.1.abstract N2 - 1073 Objectives: Gene therapy holded much promise as a potential treatment for various cardiovascular diseases. HSV1-tk gene and FIAU were the commonly used reporter gene and reporter probe to non-invasively assess location, magnitude and persistence of transgene expression in the heart and whole body. The aim of this study was to investigate the cellular uptake of [125I]-FIAU in neonate rat myocardial cells transduced HSV1-tk gene by adenoviral vector to provide the theory basis and feasibility of reporter gene imaging in heart. Methods: FAU was radiolabeled with [125I]-NaI using Iodogen iodization. The labeling efficiency, radiochemistry purity and stability were studied. Neonate rat myocardial cells were cultured in vitro and identified. An adenoviral vector carrying HSV1-tk cDNA (Ad-HSV1-tk) was constructed and transduced the myocardial cells with increasing multiplicities of infection (MOIs: 0, 4, 10, 20, 40 and 100). The transduction efficiency was identified by RT-PCR. After 24h of infection, the medium was replaced by the medium containing [125I]-FIAU. Then cellular uptake ratios were measured after 30min, 1h, 2h, 3h, 4h and 5h of incubation. Results: The labeling efficiency of [125I]-FIAU was more than 60%. The radiochemistry purity could reach 98% after purification. [125I]-FIAU was stable in PBS and in fresh human serum after 24h of labeling. The results of RT-PCR showed the expression of HSV1-tk gene in cultured myocardial cells, which suggested the susceptibility of cardiac cells to Ad-HSV1-tk. The uptake ratios of [125I]-FIAU increased with the increase of MOIs (except MOI 100, which showed lower uptake ratio than that at MOI 10) and time, which supported that FIAU uptake in cardiac cells expressing HSV1-tk gene had infect unit (IU) dependent and time dependent. Conclusions: HSV1-tk could be successfully expressed in cardiac cell in vitro using adenoviral as vector and yielded high cellular uptake of [125I]-FIAU. The results suggested that HSV1-tk, transduced by an adenoviral vector, and radiolabled FIAU as a marker gene / marker substrate could be used for reporter gene imaging in the heart. Research Support (if any): Supported by a grant from National Natural Science Foundation of China (NSFC 30400176). ER -