Angiostatin and endostatin gene therapy on murine lung metastases model utilizing cationic vector-mediated intravenous gene delivery

利用阳离子载体介导的静脉内基因递送对小鼠肺转移模型进行血管抑制素和内皮抑制素基因治疗

• 批准号: 11470276
• 负责人: FUJII Yoshitaka
• 金额: $ 11.01万
• 依托单位: NAGOYA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470276/
• 关键词: endostatin; transfection; nonviral vector; lung metastases model; antiangiogenic factor; アンジオスタチン

Inhibition of angiogenesis by production of antiangiogenic factors should be a viable approach for cancer gene therapy. Recently, nonviral vectors have been suggested as an alternative to viruses. In this study, we investigated whether intravenous administration of endostatin gene complexed with GL67/DOPE or PEI22K could inhibit the development of lung metastatic lesions in murine models. The biological activity of the expressed endostatin was also determined by its ability to prevent the growth of lung metastases using stable transfectants from murine NFSa Y83 fibrosarcoma cells expressing endostatin. RT-PCR analysis in various organs of mice after intravenous injection of endostatin gene complexed with GL67/DOPE or PEI22K showed that the highest levels of mRNA expression were found in the lung, followed by heart, spleen, kidney and liver. In addition, immunohistochemistry of whole lungs after intravenous injection of the complexes showed that transfected cells were localized in a variety of cell types, including respiratory cells. Single intravenous injection of endostatin gene complexed either GL67/DOPE or PEI22K at the time of 3 days or 7 days from fibrosarcoma cells implantation resulted in striking inhibition of formation of lung metastases after 2 weeks (compared to empty plasmid complexed each vector, 87-98% reduction in the number of lung metastases and 53-59% reduction in lung weight). These results demonstrated the potential usefulness of intravenous delivery of an antiangiogenic gene, using cationic vector-mediated gene transfer, for treatment of disseminated cancers in lung.

通过产生抗血管生成因子来抑制血管生成是肿瘤基因治疗的一种可行方法。最近，非病毒载体已被建议作为病毒的替代品。在这项研究中，我们研究了静脉注射内皮抑素基因与GL 67/DOPE或PEI 22 K复合物是否可以抑制小鼠模型肺转移性病变的发展。表达的内皮抑制素的生物活性也通过其使用来自表达内皮抑制素的鼠NFSa Y83纤维肉瘤细胞的稳定转染子防止肺转移瘤生长的能力来确定。RT-PCR分析显示，在小鼠各器官中，GL 67/DOPE或PEI 22 K复合物诱导的内皮抑素基因表达量最高，其次为心、脾、肾和肝。此外，静脉注射复合物后的整个肺的免疫组织化学显示，转染的细胞定位于多种细胞类型，包括呼吸细胞。在纤维肉瘤细胞植入后3天或7天时，单次静脉内注射与GL 67/DOPE或PEI 22 K复合的内皮抑制素基因，导致在2周后显著抑制肺转移的形成（与复合每种载体的空质粒相比，肺转移的数量减少87-98%，肺重量减少53-59%）。这些结果证明了静脉内递送抗血管生成基因，使用阳离子载体介导的基因转移，用于治疗播散性肺癌的潜在有用性。