ONCOLYTIC ADENOVIRUSES--CERVICAL CANCER GENE THERAPY

溶瘤腺病毒——宫颈癌基因治疗

• 批准号: 6137721
• 负责人: ANDRE Michael LIEBER
• 金额: $ 21.27万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-18 至 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6137721
• 关键词: Adenoviridae; DNA replication; HeLa cells; adeno associated virus group; apoptosis; athymic mouse; carcinoma; cervix neoplasms; cytolysis; gene expression; gene therapy; human papillomavirus; liver neoplasms; neoplasm /cancer genetics; neoplasm /cancer remission /regression; neoplasm /cancer therapy; nonhuman therapy evaluation; reporter genes; transfection /expression vector; tumor necrosis factor alpha; virus related neoplasm /cancer; virus replication; virus virus interaction

DESCRIPTION: (from the abstract) The investigator writes that the majority of malignant tumors are resistant to radio- or chemotherapy in advanced stages mainly due to loss of p53 function. New tumor gene therapy approaches must consider targeting every cell in the primary tumor and all metastases with a maximal cytotoxic index for cancer cells. First generation adenovirus (Ad) vectors are attractive for cancer therapy because of their potential to transduce all tumor sites after systemic application. These vectors are deleted in the E1 region, rendering them highly replication-defective. This proposal is based on the observation that E1-deleted Ad vectors replicate selectively in HPV associated cervical carcinoma cells because the HPV E6/E7 proteins which are responsible for the maintenance of the malignant phenotype effectively complement the deleted Ad E1 proteins. The goal is to develop oncolytic Ad vectors for cervical cancer that replicate in tumor-specifically and disseminate throughout the tumor, selectively killing all tumor cells by viral cytolysis or p53-independent apoptosis. The studies will be performed in immunodeficient mice with hepatic tumors derived from cervical carcinoma cell lines or in immunocompetent mice with HPV induced tumors. The specific aims are: 1. To develop an expression system based on a unique Ad-AAV hybrid vector, which activates a tumor-specific promoter only upon viral DNA replication. 2. To test whether virus dissemination throughout the tumor can be obtained if virus release from infected tumor cells is supported by cytolysis or apoptosis induced after viral replication is completed. For this purpose the investigators will use the AD-AAV system and investigate virus spread and anti-tumor efficacy after a) E3-11.6K expression, and b) expression of a transdominant i-kB mutant to sensitize tumor cells to apoptosis induced by TNF treatment or TRAIL expression. 3. To investigate the effect of antiviral immune responses on oncolytic vector spread and, if required, to suppress these responses by expression of Ad E3 proteins and/or transient immuno-modulation. These studies will give valuable information about the influence of apoptosis and host immune responses on replication-competent Ad vectors developed for tumor gene therapy. This proposal may provide a means for treatments of cervical carcinoma and has potential application for other malignancies with deregulated pRb/p16 functions that allow for replication of E1-deleted Ad vectors.

描述：（从摘要中）研究人员写道，大多数 晚期恶性肿瘤对放疗或化疗有抵抗力 主要是由于p53功能丧失所致。新的肿瘤基因治疗方法必须 考虑针对原发肿瘤和所有转移灶中的每个细胞 癌细胞的最大细胞毒性指数。第一代腺病毒（Ad） 载体对于癌症治疗很有吸引力，因为它们具有潜在的 全身应用后转导所有肿瘤部位。这些向量被删除 在 E1 区域，使它们高度复制缺陷。这个提议是 基于 E1 删除的 Ad 载体选择性复制的观察 HPV 相关的宫颈癌细胞是因为 HPV E6/E7 蛋白 负责有效维持恶性表型 补充删除的 Ad E1 蛋白。目标是开发溶瘤广告 宫颈癌载体可在肿瘤中特异性复制， 传播到整个肿瘤，通过病毒选择性地杀死所有肿瘤细胞 细胞溶解或不依赖 p53 的细胞凋亡。研究将在 患有源自宫颈癌细胞的肝肿瘤的免疫缺陷小鼠 品系或具有 HPV 诱导肿瘤的免疫活性小鼠。具体目标 是： 1. 开发基于独特的Ad-AAV杂交载体的表达系统， 它仅在病毒 DNA 复制时激活肿瘤特异性启动子。 2. 为了测试是否可以获得病毒在整个肿瘤中的传播，如果 细胞溶解或细胞凋亡支持病毒从受感染的肿瘤细胞中释放 病毒复制完成后诱导。为此目的 调查人员将使用 AD-AAV 系统来调查病毒传播和 a) E3-11.6K 表达和 b) a 表达后的抗肿瘤功效 反式显性 i-kB 突变体使肿瘤细胞对 TNF 诱导的细胞凋亡敏感 治疗或 TRAIL 表达。 3. 探讨抗病毒免疫效果 对溶瘤载体传播的反应，并在需要时抑制这些 通过表达 Ad E3 蛋白和/或瞬时免疫调节来做出反应。 这些研究将提供有关细胞凋亡影响的有价值的信息 和宿主对具有复制能力的 Ad 载体的免疫反应 肿瘤基因治疗。该提案可能提供一种治疗方法 宫颈癌，并具有治疗其他恶性肿瘤的潜在应用 解除管制的 pRb/p16 功能，允许复制 E1 删除的 Ad 向量。