ONCOLYTIC ADENOVIRUSES--CERVICAL CANCER GENE THERAPY

溶瘤腺病毒——宫颈癌基因治疗

• 批准号: 6626632
• 负责人: ANDRE Michael LIEBER
• 金额: $ 23.25万
• 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 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6626632
• 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：（摘要）调查人员写道大多数 恶性肿瘤在晚期对放射性或化学疗法有抵抗力 主要是由于p53功能的损失。新的肿瘤基因治疗方法必须 考虑靶向原发性肿瘤中的每个细胞和所有转移 癌细胞的最大细胞毒性指数。第一代腺病毒（AD） 载体对癌症治疗具有吸引力，因为它们的潜力 全身应用后转导所有肿瘤部位。这些向量已删除 在E1区域，使它们高度复制缺陷。该提议是 基于这样的观察，e1删除的ad载体选择性地在 HPV相关的宫颈癌细胞，因为HPV E6/E7蛋白 有效地维持恶性表型 补充已删除的AD E1蛋白。目标是开发溶瘤广告 宫颈癌的载体，在特定于肿瘤中复制和 在整个肿瘤中传播，通过病毒选择性杀死所有肿瘤细胞 细胞解剖或p53非依赖性凋亡。研究将在 免疫缺陷小鼠，肝癌细胞衍生的肝肿瘤 线或具有HPV诱导肿瘤的免疫能力小鼠。具体目标 为：1。要基于唯一的AD-AAV混合矢量开发表达系统， 仅在病毒DNA复制后激活肿瘤特异性启动子。 2。 测试是否可以在整个肿瘤中传播病毒 从感染的肿瘤细胞中释放病毒由细胞溶解或凋亡支持 病毒复制完成后诱导。为此目的 调查人员将使用AD-AAV系统并研究病毒传播， a）E3-11.6K表达后的抗肿瘤功效，b）表达 I-KB突变体的跨性I-KB突变体使肿瘤细胞对TNF诱导的细胞凋亡敏感 治疗或跟踪表达。 3。研究抗病毒免疫的作用 关于溶瘤载体的响应，并在需要时抑制这些 通过表达AD E3蛋白和/或瞬时免疫调节的反应。 这些研究将提供有关凋亡影响的宝贵信息 并在为复制能力的AD矢量上产生免疫反应 肿瘤基因治疗。该提案可以提供一种治疗方法 宫颈癌，并有潜在适用于其他恶性肿瘤 放松管制的PRB/P16功能，可以复制E1删除AD 向量。