Gene Therapy for Human Lung Adenocarcinoma Using a Suicide Gene Driven by a Lung-Specific Promoter Delivered by JC Virus-Like Particles.
Lung adenocarcinoma, the most commonly diagnosed type of lung cancer, has a poor prognosis even with combined surgery, chemotherapy, or molecular targeted therapies. Most patients are diagnosed with an in-operable advanced or metastatic disease, both pointing to the necessity of developing effective therapies for lung adenocarcinoma. Surfactant protein B (SP-B) has been found to be overexpressed in lung adenocarcinoma. In addition, it has also been demonstrated that human lung adenocarcinoma cells are susceptible to the JC polyomavirus (JCPyV) infection. Therefore, we designed that the JCPyV virus-like particle (VLP) packaged with an SP-B promoter-driven thymidine kinase suicide gene (pSPB-tk) for possible gene therapy of human lung adenocarcinoma. Plasmids expressing the GFP (pSPB-gfp) or thymidine kinase gene (pSPB-tk) under the control of the human SP-B promoter were constructed. The promoter's tissue specificity was tested by transfection of pSPB-gfp into A549, CH27, and H460 human lung carcinoma cells and non-lung cells. The JCPyV VLP's gene transfer efficiency and the selective cytotoxicity of pSPB-tk combined with ganciclovir (GCV) were tested in vitro and in a xenograft mouse model. In the current study, we found that SP-B promoter-driven GFP was specifically expressed in human lung adenocarcinoma (A549) and large cell carcinoma (H460) cells. JCPyV VLPs were able to deliver a GFP reporter gene into A549 cells for expression. Selective cytotoxicity was observed in A549 but not non-lung cells that were transfected with pSPB-tk or infected with pSPB-tk-carrying JCPyV VLPs. In mice injected with pSPB-tk-carrying JCPyV VLPs through the tail vein and treated with ganciclovir (GCV), a potent 80% inhibition of growth of human lung adenocarcinoma nodules resulted. The JCPyV VLPs combined with the use of SP-B promoter demonstrates effectiveness as a potential gene therapy against human lung adenocarcinoma.
Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan.|Department of Pediatrics, Ditmanson Medical Foundation Chiayi Christian Hospital, Chiayi, Taiwan.
Chun-Nun
CN
Chao
Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan.|Department of Urology, Ditmanson Medical Foundation Chiayi Christian Hospital, Chiayi, Taiwan.
Mien-Chun
MC
Lin
Department of Medical Research, Ditmanson Medical Foundation Chiayi Christian Hospital, Chiayi, Taiwan.
Chiung-Yao
CY
Fang
Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan.
Pei-Lain
PL
Chen
Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan.
Deching
D
Chang
Department of Medical Research, Ditmanson Medical Foundation Chiayi Christian Hospital, Chiayi, Taiwan.|Department of Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan.
Cheng-Huang
CH
Shen
Department of Microbiology and Immunology, Chung Shan Medical University, Taichung, Taiwan.
Meilin
M
Wang
D000072283:A549 Cells; D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D000077192:Adenocarcinoma of Lung; D016923:Cell Death / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D015774:Ganciclovir / Q000494:pharmacology; D017930:Genes, Reporter; D041722:Genes, Transgenic, Suicide; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007577:JC Virus / Q000378:metabolism*; D008168:Lung / Q000378:metabolism*; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D009928:Organ Specificity / Q000187:drug effects; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D037701:Pulmonary Surfactant-Associated Protein B / Q000378:metabolism; D013937:Thymidine Kinase / Q000378:metabolism; D014162:Transfection; D014771:Virion / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays
D037701:Pulmonary Surfactant-Associated Protein B; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D013937:Thymidine Kinase; D015774:Ganciclovir