Strategies to Enhance Adenoviral Mediated Tumor Killing

简介(申请人提供)：肺癌是导致癌症的主要原因 美国男性和女性的死亡人数，只有13%的肺部疾病患者 癌症存活5年。非小细胞肺癌的支持性护理， 是最常见的一种，中位存活率仅为4个月。新的 联合用药只能将存活率提高到8-10个月。新技术的发展 因此，治疗方式非常重要。这项研究详细载于 这项建议旨在开发新的、高效但安全的基因疗法。 治疗肺癌的方法。该方法将基于修改 针对肺癌病毒复制和细胞杀伤的腺病毒基因 细胞。细胞周期调控基因如P53和Rb在 在癌细胞和腺病毒感染的细胞中。腺病毒基因的缺失 因此，改变对细胞周期的控制可能以病毒复制为目标 癌细胞。一种Elb-55kD缺失病毒已被引入靶向p53 然而，一些报告对这种方法提出了质疑。在……里面 这项建议将确定带有修饰的ela基因的腺病毒 无法灭活Rb和/或p300，将以病毒复制为目标 癌细胞。为了进一步将病毒复制到肺癌细胞， 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进 复制型腺病毒载体的溶瘤活性。腺病毒 E1b-19kD蛋白是一种有效的细胞凋亡抑制因子，是一种具有 这种基因被删除，更有效地杀死并通过单层 肿瘤细胞。因此，E1b-19kD缺失的影响现在将是 在小鼠模型中进行了评估。此外，结合病毒的效果 将对细胞毒剂的感染进行评估。在第三个目标中， 结合的腺病毒载体的溶瘤活性和特异性 E1b-19kD基因修饰的ELA基因的转录靶向 将对删除进行评估。安全方面很难在小鼠身上进行评估 与人腺病毒一样的模型不能在小鼠细胞中复制。在第四个目标中， 这些腺病毒载体的安全性将通过测量 新鲜分离株病毒复制、细胞杀伤和诱导细胞凋亡 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.