Adenoviral herpes simplex virus thymidine kinase gene therapy in an orthotopic lung cancer model
Adenoviral herpes simplex virus thymidine kinase gene therapy in an orthotopic lung cancer model
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DOI：
10.1016/s0003-4975(02)03572-5
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发表时间：
2002-06-01
影响因子：
4.6
通讯作者：
Kosai, K
中科院分区：
文献类型：
--
作者：
Fukunaga, M;Takamori, S;Kosai, K
背景资料。虽然腺病毒介导的单纯疱疹病毒胸苷激酶(HSVtk)基因治疗是一种新的有效治疗肺癌的潜在候选方法，但以往的研究仅在裸鼠皮下肿瘤模型中进行。我们使用不能胜任的小鼠，在更具临床相关性的原位肺癌模型中，研究了治疗潜力与准确的腺病毒基因转导效率的相关性。为分析腺病毒载体HSVtk基因转导和更昔洛韦的细胞毒作用，采用体外细胞增殖实验。以预定的基因转导效率，将预先感染的同种小鼠肺癌细胞以预定的基因转导效率接种于肺内，对免疫功能正常的原位肺癌小鼠进行存活研究。体外感染携带HSVtk基因的腺病毒并加入更昔洛韦可有效杀伤肿瘤细胞。体内实验中，对照组小鼠均于接种肿瘤后26天内死于原发肺癌的快速生长和纵隔淋巴结转移。相比之下，在注射更昔洛韦后，分别有50%和100%的小鼠在接种中高表达HSVtk的腺病毒感染的肿瘤细胞后存活超过40天。基因转导效率为67%。腺病毒介导的HSVtk基因治疗在获得基因转导效率和足够的HSVtk表达水平的情况下，可能对肺癌具有治疗作用。此外，目前的发现强调了小鼠原位肺癌模型对基因治疗研究的重要性。
Background. Although adenovirus-mediated herpes simplex virus thymidine kinase (HSVtk) gene therapy is a potential candidate for a novel effective therapy for lung cancer, previous studies have been performed only in a subcutaneous tumor model employing nude mice. We studied therapeutic potentials in correlation with accurate adenoviral gene transduction efficiency in a more clinically relevant orthotopic lung cancer model employing imunocompetent mice.Methods. To analyze the cytotoxicity of adenovirad HSVtk gene transduction and ganciclovir, a cell proliferation assay was performed in vitro. A survival study was carried out in immunocompetent mice with orthotopic lung cancer, which was generated by intrapulmonary inoculation with syngeneic murine lung cancer cells that had been infected beforehand with each adenoviral vector at the predetermined gene transduction efficiencies.Results. Tumor cells were efficiently killed by infection with adenovirus carrying the HSVtk gene with the addition of ganciclovir in vitro. In the in vivo experiment all control mice died of rapid growth of the primary lung cancer and of metastases to mediastinal lymph nodes within 26 days after tumor inoculation. In contrast 50% and 100% of mice survived more than 40 days after inoculation with adenovirally HSVtk-transfected tumor cells that moderately and highly expressed HSVtk, respectively, when followed by ganciclovir administration. Gene transduction efficiencies were 67%.Conclusions. Adenovirus-mediated HSVtk gene therapy may be therapeutic for lung cancer when gene transduction efficiencies and sufficient expression levels of HSVtk can be achieved. Moreover, the present findings underscore the importance of the mouse orthotopic lung cancer model for studies of gene therapy.