Preclinical studies of a new generation retroviral vector for ovarian cancer BRCA1 gene therapy.
Preclinical studies of a new generation retroviral vector for ovarian cancer BRCA1 gene therapy.
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新一代逆转录病毒载体用于卵巢癌BRCA1基因治疗的临床前研究。
DOI：
10.1006/gyno.2000.5969
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Holt,JT
中科院分区：
文献类型：
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作者：
Tait,DL;Obermiller,PS;Holt,JT
目的本研究的目的是确定第二代补体抗性逆转录病毒BRCA1载体MFG-BRCA1用于卵巢癌基因治疗的临床前稳定性、毒性和疗效。方法MFG-BRCA1包装在人293肾细胞中，在cGMP条件下生产和测试，并经美国食品和药物管理局允许用于人体。通过 PCR 分析在小鼠和人血清中进行载体稳定性研究。在尸检时评估动物的毒性，评估炎症和器官损伤的组织学迹象。对卵巢癌细胞和乳腺癌细胞进行了组织培养功效研究。在雌性 nu/nu 小鼠中进行动物功效研究。小鼠腹腔注射SKOV-3卵巢癌细胞，让肿瘤生长4周。用 MFG-BRCA1 或对照载体腹膜内治疗小鼠。比较 MFG-BRCA1 与对照组的动物存活率。结果 MFG-BRCA1 在人血清中比 LXSN-BRCA1sv 更稳定。炎症性腹膜炎所证明的毒性很小。与单独使用 MFG 载体相比，在两种细胞系中使用 MFG-BRCA1 获得的克隆显着减少。在动物中进行的 MFG-BRCA1 功效研究表明，与对照载体相比，存活率提高了近三倍，与第一代 LXSN-BRCA1sv 载体相比，存活率提高了两倍。 结论 重新设计的补体抗性 MFG-BRCA1 逆转录病毒载体比上一代 LXSN-BRCA1sv 载体更有效、更稳定。
ObjectiveThe aim of this study was to determine the preclinical stability, toxicity, and efficacy of a second-generation complement-resistant retroviral BRCA1 vector, MFG-BRCA1, for ovarian cancer gene therapy.MethodsMFG-BRCA1 was packaged in human 293 renal cells and manufactured and tested under cGMP conditions and is allowed for use in humans by the Food and Drug Administration. Vector stability studies were performed in mice and human serum by PCR analysis. Toxicity in the animals was assessed at necropsy, evaluating for histological signs of inflammation and organ damage. Tissue culture efficacy studies were performed on ovarian and breast cancer cells. Animal efficacy studies were conducted in female nu/nu mice. Mice were injected intraperitoneally with SKOV-3 ovarian cancer cells and tumors were allowed to grow for 4 weeks. Mice were treated intraperitoneally with MFG-BRCA1 or control vectors. Survival of animals was compared in the MFG-BRCA1 versus the control groups.ResultsMFG-BRCA1 was more stable in human serum than LXSN-BRCA1sv. Toxicity as demonstrated by an inflammatory peritonitis was minimal. Significantly fewer clones were obtained using the MFG-BRCA1 versus the MFG vector alone in both cell lines. Efficacy studies in animals of MFG-BRCA1 demonstrated a near threefold increase in survival over control vector and twofold increase compared to the first generation LXSN-BRCA1sv vector.ConclusionThe reengineered complement-resistant MFG-BRCA1 retroviral vector is more effective and more stable than the previous generation LXSN-BRCA1sv vector.