Strategies to Enhance Adenoviral Mediated Tumor Killing

描述（由申请人提供）：肺癌是癌症的主要原因 在美国男性和女性中，只有13%的人患有肺部疾病， 癌症存活5年。非小细胞肺癌的支持性护理， 是最常见的形式，其平均存活率仅为4个月。新 药物组合仅将存活期提高到8-10个月。发展新 因此，治疗方式非常重要。详细的研究在 该提案旨在开发新型、高效、安全的基因疗法， 治疗肺癌的方法。该方法将基于修改 腺病毒基因靶向病毒复制和细胞杀伤肺癌 细胞细胞周期调控基因如p53和Rb在细胞凋亡中均失活。 癌细胞和腺病毒感染的细胞中。腺病毒基因缺失 因此，改变细胞周期的控制可以靶向病毒复制， 癌细胞已引入Elb-55 kD缺失病毒以靶向p53 突变的癌细胞，然而，一些报告质疑这种方法。在 这项建议，将确定是否有一个修饰的Ela基因的腺病毒， 不能抑制Rb和/或p300的病毒，将靶向病毒复制， 癌细胞为了进一步将病毒复制靶向肺癌细胞， 修饰的Ela基因的转录将被限制在肺细胞中， 使用肺特异性启动子。第二个目标的重点将是改善 复制型腺病毒载体的溶瘤活性。腺病毒 E1 b-19 kD蛋白是一种有效的细胞凋亡抑制剂， 这种基因删除，更有效地杀死和传播通过一个单层的 肿瘤细胞因此，E1 b-19 kD缺失的影响现在将是 在小鼠模型中评估。此外，结合病毒 将评价细胞毒性剂的感染。在第三个目标中， 腺病毒载体的溶瘤活性和特异性，其组合 用E1 b-19 kD基因转录靶向修饰的Ela基因 将对删除进行评估。在小鼠中难以评价安全性方面 因为人腺病毒不能在小鼠细胞中复制。第四个目标， 这些腺病毒构建体安全性将通过测量 病毒复制、细胞杀伤和诱导凋亡 肺肿瘤细胞

DESCRIPTION (provided by applicant): Lung cancer is the leading cause of cancer death in American men and women, and only 13 percent of people who develop lung cancer survive 5 years. Supportive care for non-small cell lung cancer, which is the most common form, yields median survival rates of just 4 months. New drug combinations improve survival only to 8-10 months. The development of new treatment modalities is therefore of great importance. The research detailed in this proposal aims to develop novel, highly effective, but safe gene therapy approaches to treat lung cancer. The approach will be based on modifying adenoviral genes to target viral replication and cell killing to lung cancer cells. Cell cycle regulating genes such as p53 and Rb are inactivated both in cancer cells and in adenovirus infected cells. Deletion of the adenoviral genes that alter control of the cell cycle may therefore target viral replication to cancer cells. An Elb-55kD deleted virus has been introduced to target p53 mutant cancer cells, however several reports have questioned this approach. In this proposal, it will be determined if an adenovirus with a modified Ela gene that is unable to inactivate Rb and/or p300, will target viral replication to cancer cells. To further target viral replication to lung cancer cells, transcription of the modified Ela gene will be restricted to lung cells by using lung specific promoters. The focus of the second aim will be improving the oncolytic activity of a replicating adenoviral vector. The adenoviral E1b-19kD protein is a potent inhibitor of apoptosis, and an adenovirus with this gene deleted, more efficiently kills and spreads through a monolayer of tumor cells. The effects of the E1b-19kD deletion will therefore now be evaluated in a mouse model. In addition, the effects of combining viral infection with cytotoxic agents will be evaluated. In the third aim, the oncolytic activity and specificity of an adenoviral vector, that combines transcriptional targeting of the modified Ela gene with an E1b-19kD gene deletion will be evaluated. Safety aspects are difficult to evaluate in a mouse model as human adenoviruses do not replicate in mouse cells. In the fourth aim, the safety of these adenoviral constructs will be evaluating by measurement of viral replication, cell killing and induction of apoptosis in freshly isolated lung tumor cells.