In vivo electroporation (EP) is an efficient method for effective gene transfer and is highly expected for application in anticancer gene therapy. Non-invasive monitoring of gene transfer/expression is critical for optimal gene therapy. Here we report in vivo optical and high-field magnetic resonance imaging (MRI) of EP-mediated transgene expression in a tumor model. Initially, we observed spatio-temporal change in in vivo EP-mediated transgene expression by optical imaging using red fluorescence protein (RFP) as a reporter gene. Next, we constructed a dual-reporter plasmid carrying a gene-encoding MRI reporter ferritin heavy chain and RFP gene to visualize the intratumoral transgene expression by dual modality. Cells transfected with this plasmid showed lower signal intensity on in vitro T2-weighted cellular MRI and quantitatively increased the transverse relaxation rate (1/T2) compared with control cells. After conducting in vivo EP in an experimental tumor, the plasmid-injected region showed both fluorescent emissions in optical imaging and detectably lowered signal on T2-weighted MRI. The correlative immunohistological findings confirmed that both the reporter transgenes were co-expressed in this region. Thus, our strategy provides a platform for evaluating EP-mediated cancer gene therapy easily and safely without administering contrast agent or substrate.
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We thank Dr Paolo Arosio (University of Brescia, Italy) for the generous gift of ferritin cDNA; Dr Jun-ichi Miyazaki (Osaka University, Japan) for the generous donation of pCAGGS vector; Kosho Suzuki (NEPA GENE) for technical suggestions for in vivo EP; Michiko Narazaki, Misao Yoneyama, Sayaka Shibata and Shigeyoshi Saito for skillful assistance with the MRI experiments; Hikaru Takenaka for preparation of the paper; members in our group for helpful discussions; Drs Yasuhito Sasaki, Yoshiharu Yonekura and Shuji Tanada for encouragements for this study. This study was supported in part by a Grant-in-Aid for Scientific Research for Exploratory Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan (18659357: S Hasegawa); Intramural Research Grant from the National Institute of Radiological Sciences (S Hasegawa).
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Aung, W., Hasegawa, S., Koshikawa-Yano, M. et al. Visualization of in vivo electroporation-mediated transgene expression in experimental tumors by optical and magnetic resonance imaging. Gene Ther 16, 830–839 (2009). https://doi.org/10.1038/gt.2009.55
- in vivo electroporation
- transgene expression
- in vivo optical imaging
- magnetic resonance imaging
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