Oncolytic viral therapy for disseminated peritoneal ovarian cancer using a novel replication-competent herpes simplex virus type 1 mutant in mice.

使用新型具有复制能力的 1 型单纯疱疹病毒突变体对小鼠进行溶瘤病毒治疗，用于治疗播散性腹膜卵巢癌。

• 批准号: 13671760
• 负责人: NAWA Akihiro
• 金额: $ 2.37万
• 依托单位: Aichi Cancer Center
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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/grant/KAKENHI-PROJECT-13671760/
• 关键词: replication-competent; herpes simplexvirus type1 mutant; ovarian cancer; peritoneally disseminated neoplasm; mouse model; syncytial formation; apoptosis; immuno-competent mouse; 増殖型HSVI変異株

We prepared two attenuated mutant HSV-1 strains. One is an HSV-1 mutant, hrR3, and another is a new replication competent HSV-1 mutant, HR522 ; this virus, expressing the lacZ reporter gene, induces syncytium formation in infected cells. We compared the efficacy of HR522 with Taxol and hrR3 in the treatment of nude mice harboring human ovarian cancer cells. We also examined the effect of the prodrug ganciclovir (GCV) on the treatment mediated by these HSVs. The survival of mice treated with a high titer hrR3 (5x10^7 plaque-forming units ; PFU) was significantly prolonged as compared to the group given Taxol (p<0.0001 ; Log-Rank test). Although the survival of mice treated with high titer HR522 (5x10^7 PFU) was not significantly prolonged compared with Taxol-treated group (p=0.212 ; Log-Rank test), GCV markedly enhanced the efficacy of HR522 administration (p<0.005 ; vs Taxol ; Log-Rank test). The lacZ gene product, visualized using 5-brorno-4-chrolo-3-indoryl-β-D- galactopyranoside (X-gal) histochemistry, was detected in HR522-treated tumors in areas also exhibiting apoptotic changes. Another study demonstrated that a clonal derivative of HSV-1 strain HF done 10 very effectively treated peritoneally disseminated neoplasm in an immunocompetent animal model and that all of survived mice acquired resistance to rechallenge of tumor cells. HF clone 10 induces syncytia formation in vitro. A sequential administration ofHF clone 10 attained a long-term survival over 90 days after tumor injection in 8 of the 9 treated mice without any signs of diseases. The results suggested that treatment of peritoneally disseminated tumor with HF clone 10 induced a specific anti-tumor immune response. We found that clone 10 has a deletion of 39kbp in the right end of UL and UL/IRL junction, resulting in the loss of UL 56 expression. Moreover, a 23kbp deletion and extensive rearrangement were observed in the left end of the genome.

我们制备了两株HSV-1减毒突变株。一个是HSV-1突变体hrR3，另一个是新的具有复制能力的HSV-1突变体HR522；这种病毒表达LacZ报告基因，在感染细胞中诱导合胞体形成。我们比较了HR522、紫杉醇和hrR3对荷人卵巢癌细胞裸鼠的治疗效果。我们还检测了前药更昔洛韦(GCV)对这些HSV介导的治疗的影响。与紫杉醇组相比，高滴度hrR3(5x10^7空斑形成单位；pfu)组小鼠的存活时间显著延长(p&lt；0.0001；Log-Rank检验)。与紫杉醇治疗组相比，高滴度HR522(5×10~(-7)pfu)组小鼠的存活时间没有显著延长(p=0.212；对数等级检验)，但GCV显著增强了hR_(522)给药的疗效(p&lt；0.005；vs紫杉醇；对数等级检验)。用5-溴-4-氯-3-吲哚-β-D-半乳糖苷(X-GAL)组织化学显影的LacZ基因产物，在HR522治疗的肿瘤中也被检测到，在也表现出凋亡变化的区域。另一项研究表明，HSV-1株HF的克隆衍生物在免疫活性动物模型中对腹膜播散性肿瘤进行了非常有效的治疗，所有存活的小鼠都获得了对肿瘤细胞再次攻击的抵抗力。HF克隆10在体外诱导合胞体形成。在9只接受治疗的小鼠中，有8只在注射肿瘤后连续注射HF克隆10获得了超过90天的长期存活，没有任何疾病迹象。结果提示，HF克隆10治疗腹膜播散性肿瘤可诱导特异性抗肿瘤免疫反应。我们发现克隆10在UL和UL/IRL连接的右端有39kbp的缺失，导致UL 56的表达缺失。此外，在基因组的左端还观察到23kbp的缺失和广泛的重排。

项目成果

Nicolson GL, Nawa A, et al.: "Tumor metastasis-associated human MTA1 gene and its MTA1 protein product: role in epithelial cancer cell invasion, proliferation and nuclear regulation"Clinical experimental metastasis. (in press).

Nicolson GL、Nawa A等人：“肿瘤转移相关的人MTA1基因及其MTA1蛋白产物：在上皮癌细胞侵袭、增殖和核调节中的作用”临床实验转移。

Takakuwa H, Nawa A, Nishiyama Y, et al.: "Oncolytic viral therapy using a spontaneously generated herpes simplex virus type 1 variant for peritoneally disseminated tumor in immunocompetent mice"Archives of Virology. (in press).

Takakuwa H、Nawa A、Nishiyama Y 等人：“使用自发产生的单纯疱疹病毒 1 型变体进行溶瘤病毒疗法，用于免疫活性小鼠的腹膜播散性肿瘤”病毒学档案。