Development of EBV/polyplex as gene therapy against prostate cancer

开发 EBV/polyplex 作为前列腺癌基因疗法

• 批准号: 13470339
• 负责人: NAKAO Masahiro
• 金额: $ 6.85万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470339/
• 关键词: prostate cancer; EBV-based plasmid vector; Fas ligand; EBV; polyplex; lipoplex; 腎細胞癌; 選択的COX-2阻害剤; 抗Fas抗体; JTE-522; 遺伝子治療; EBV-プラスミドベクター; Fas; アポトーシス; cationic polymer

To accomplish efficient nonviral gene therapy against prostate cancer (PC), Epstein-Barr virus (EBV)-based plasmid vectors containing EBNA1 gene and oriP were employed and combined with a cationic polymer or cationic lipid. When EBV-plasmid/poly-amidoamine dendrimer complex was injected into PC-3-derived tumors established in severe combined immunodeficiency mice, a considerable expression of marker gene was obtained in the tumors, and the expression level was more than eight-fold higher than that achieved by conventional plasmid vector/dendrimer. Since most PC cells express the apoptotic signal molecule Fas (Apo-1/CD95) on their surface, Fas ligand (FasL) gene was transferred into PC cells to kill the tumor cell. In vitro transfection with pGEG.FasL (an EBV-plasmid with the FasL gene) significantly reduced the viability of PC cells, which subsequently underwent apoptosis. Intratumoral injection of pGEG.FasL into PC induced significant growth suppression of the xerograft tumors, in which typical characteristics of apoptosis were demonstrated by TUNEL staining and electron microscopic observations. When pGEG.FasL transfer was accompanied by systemic administrations of cisplatin, the tumors were inhibited even more remarkably, leading to prolonged survival of the animals. FasL gene transfection by means of EBV-based plasmid/cationic macromolecule complexes may provide a practical therapeutic strategy against PC.

为了实现前列腺癌(PC)的有效非病毒基因治疗，以EB病毒(EBV)为基础的含有EBNA1基因和ORIP基因的质粒载体与阳离子聚合物或阳离子脂质结合在一起。将EBV-质粒/多胺树状大分子复合体注射到严重联合免疫缺陷小鼠PC-3来源的肿瘤中，肿瘤中有大量标志物基因的表达，其表达水平是常规载体/树突状大分子的8倍以上。由于大多数PC细胞表面表达凋亡信号分子Fas(Apo-1/CD95)，因此将Fas配体(FasL)基因导入PC细胞以杀伤肿瘤细胞。在体外，pGEG.FasL(一种带有FasL基因的EBV-质粒)显著降低了PC细胞的活力，随后PC细胞发生了凋亡。PC瘤内注射pGEG.FasL可明显抑制移植瘤的生长，TUNEL染色和电子显微镜观察显示移植瘤呈现典型的细胞凋亡特征。当pGEG.FasL转移伴随顺铂全身给药时，肿瘤被抑制得更加明显，从而延长了动物的存活时间。以EB病毒为载体的质粒/阳离子大分子复合体转导FasL基因可能为PC提供一种实用的治疗策略。

项目成果

Kishida T. et al.: "In vivo electroporation-mediated transfer of interleukin-12 and interleukin-18 genes induces significant antitumor effects against melanoma in mice."Gene Therapy. 8. 1234-1240 (2001)

Kishida T. 等人：“体内电穿孔介导的白细胞介素 12 和白细胞介素 18 基因转移可诱导小鼠体内针对黑色素瘤的显着抗肿瘤作用。”基因治疗。

Asada H, et al.: "Significant antitumor effects obtained by autologous tumor cell vaccine enginerered to secrete interleukin (IL)-12 abd IL-18 by means of the EBV/lipoplex"Molecular Therapy. 5. 609-616 (2002)

Asada H 等人：“通过 EBV/lipoplex 的方式，将自体肿瘤细胞疫苗设计为分泌白细胞介素 (IL)-12 和 IL-18，获得了显着的抗肿瘤效果”分子疗法。

Nakanishi H, et al.: "Significant Antitumoral Activity of Cationic Multilamellar Liposomes Containing Human IFN-β Gene against Human Renal Cell Carcinoma"Clinical Cancer Research. 9-3. 1129-1135 (2003)

Nakanishi H等人：“含有人IFN-β基因的阳离子多层脂质体对人肾细胞癌的显着抗肿瘤活性”，临床癌症研究9-3(2003)。

Nakanishi H, et al.: "Nonviral genetic transfer of Fas ligand induced significant growth suppression and apoptotic tumor cell death in prostate cancer in vivo"Gene Therapy. 10・5. 434-442 (2003)

Nakanishi H 等人：“Fas 配体的非病毒遗传转移在体内诱导前列腺癌中显着的生长抑制和凋亡肿瘤细胞死亡”基因治疗 10·5。

Kishida T, et al.: "In vivo electroporation-mediated transfer of interleukin-12 and interleukin-18 genes induces significant antitumor effects against melanoma in mice"Gene Therapy. 8. 1234-1240 (2001)

Kishida T 等人：“体内电穿孔介导的白细胞介素 12 和白细胞介素 18 基因转移可诱导小鼠体内针对黑色素瘤的显着抗肿瘤作用”基因疗法。