E2F-1 Cancer Gene Therapy

E2F-1癌症基因治疗

• 批准号: 6797897
• 负责人: Kelly M McMasters
• 金额: $ 24.44万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6797897
• 关键词: CD95 molecule; antineoplastics; apoptosis; biological signal transduction; camptothecin; cell growth regulation; cell line; combination therapy; doxorubicin; drug resistance; gene therapy; genetic transduction; mitogen activated protein kinase; neoplasm /cancer chemotherapy; neoplasm /cancer genetics; neoplastic cell; northern blottings; temozolomide; transcription factor; western blottings

DESCRIPTION (PROVIDED BY APPLICANT): The major reason for the failure of anticancer drug therapy in most advanced solid tumors is the presence of chemotherapy-resistant tumor cells. Apoptosis is now considered to be an essential event in antitumor drug induced death. Therefore, cells with deficient apoptotic pathways may be incapable of producing an efficient apoptotic response, regardless of the agent to which they are exposed. For this reason, gene therapy using proapoptotic genes that can either trigger apoptosis in chemo-resistant cells or reconstitute the apoptotic pathway to enhance chemotherapy is a potential therapeutic strategy. The transcription factor, E2F-1, has recently been identified as a potent tumor suppressor. We and others have shown that E2F- 1 overexpression efficiently induces apoptosis in a variety of tumor types, independent of p53 status, both in vitro and in vivo. However, the mechanisms by which E2F-1 induces apoptosis remain ill defined. Mechanistically, our preliminary data implicate activation of the Fas death receptor pathway, involving ASK1, JNK, p38, and ERK MAP kinases in E2F- 1-mediated apoptosis, a novel finding. Furthermore, NFkappaB activation and downregulation of inhibitors of apoptosis (IAPs, specifically XIAP and cIAP-1) and the antiapoptotic Bcl-2 family member, Mcl-1 were seen after E2F-1- overexpression, again novel findings. Furthermore, our preliminary studies indicate that topoisomerase I and II agents, and the drug temozolomide work cooperatively with E2F-1 to induce apoptosis in colon cancer and melanoma cells in vitro and in vivo. Based on substantial preliminary data and the existing literature, we have proposed a model of the signal transduction pathways involved in E2F-1-mediated apoptosis. To evaluate this model and further dissect the molecular mechanisms of E2F-1-induced apoptosis, SK-MEL-2 melanoma and HT-29 colon cancer cells will be studied. Following adenovirus-mediated gene transfer we will analyze the role of Fas/FasL interaction, JNK, p38, and ERK activation, NFKB activation, and XIAP, cIAP-1 and Mcl-1 in E2F-1-mediated apoptosis. Investigation will also be carried out using combinations of adenovirus E2F-1 and specific chemotheraeutic agents that act cooperatively with E2F-1 (adriamycin. camptothecin, temozolomide) to evaluate the mechanisms by which E2F-1 augments the apoptotic response to chemotherapy. Finally, a novel adenoviral vector expressing a truncated E2F-1 mutant that maintains apoptotic function but lacks E2F-1 transactivation function will be constructed and tested for potential enhancement of tumor death. These studies will enhance our understanding of E2F-1-mediated cell death and facilitate the design of improved treatment strategies.

描述（申请人提供）：失败的主要原因 大多数晚期实体瘤的抗癌药物治疗是存在 耐化疗的肿瘤细胞。细胞凋亡现在被认为是一种 抗肿瘤药物诱导死亡的基本事件。因此，具有 缺乏凋亡途径可能不能产生有效的 细胞凋亡反应，无论其暴露于何种试剂。为此 原因是，使用促凋亡基因的基因治疗， 或重建凋亡途径，以增强 化疗是一种潜在的治疗策略。转录因子， E2 F-1，最近被鉴定为有效的肿瘤抑制剂。我们和其他人 已经表明E2 F- 1过表达有效地诱导细胞凋亡， 多种肿瘤类型，独立于p53状态，在体外和体内。 然而，E2 F-1诱导细胞凋亡的机制仍然不清楚。 从机制上说，我们的初步数据暗示Fas死亡的激活 受体途径，涉及E2 F-1中的ASK 1、JNK、p38和ERK MAP激酶。 1-介导的凋亡，一个新的发现。此外，NF κ B活化和 下调细胞凋亡抑制剂（IAP，特别是XIAP和cIAP-1） 而抗凋亡Bcl-2家族成员Mcl-1在E2 F-1-E2 F-1-E2 F-1-E2 F-1后表达。 过度表达，再次发现新的发现。此外，我们的初步研究 表明拓扑异构酶I和II试剂以及药物替莫唑胺起作用 与E2 F-1协同诱导结肠癌和黑色素瘤细胞凋亡 在体外和体内。根据大量的初步数据和现有的 文献中，我们提出了一个模型的信号转导途径 参与E2 F-1介导的细胞凋亡。为了评估这个模型， 分析E2 F-1诱导细胞凋亡、SK-MEL-2黑色素瘤的分子机制 和HT-29结肠癌细胞进行研究。在腺病毒介导的 基因转移，我们将分析Fas/FasL相互作用，JNK，p38， E2 F-1介导的细胞凋亡中ERK激活、NF κ B激活和XIAP、cIAP-1和Mcl-1的表达 凋亡还将使用以下组合进行调查： 腺病毒E2 F-1和协同作用的特异性化疗剂 用E2 F-1（阿霉素。喜树碱，替莫唑胺），以评估机制 E2 F-1通过其增强对化疗的凋亡反应。最后 表达截短的E2 F-1突变体的新型腺病毒载体 将构建具有凋亡功能但缺乏E2 F-1反式激活功能的 并测试肿瘤死亡的潜在增强。这些研究将提高 我们对E2 F-1介导的细胞死亡的理解， 改善治疗策略。