Expression of the TRAIL Gene for Cancer Therapy

TRAIL 基因的表达用于癌症治疗

• 批准号: 6603790
• 负责人: BINGLIANG FANG
• 金额: $ 29.82万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6603790
• 关键词: Adenoviridae; RNA directed DNA polymerase; antineoplastics; apoptosis; athymic mouse; cell line; colon neoplasms; fusion gene; gene expression; gene therapy; genetic promoter element; green fluorescent proteins; histopathology; human tissue; ligands; liver cells; neoplasm /cancer therapy; neoplastic cell; nonhuman therapy evaluation; recombinant proteins; technology /technique development; telomerase; transfection /expression vector; tumor necrosis factor alpha; tumor promoters

Because of their ability to elicit cell death in a variety of tumor cells but not in normal cells, recombinant proteins of the TNFalpha-related-apoptosis-inducing ligand (TRAIL) appear to be promising medicine for cancer patients. However, a recent finding of apoptosis induction by recombinant TRAIL protein in primary normal human hepatocytes raised the alarm about the possibility of serious toxicity when the TRAIL proteins are administered systemically. Our preliminary study also showed that expression of the full-length TRAIL protein in primary human hepatocytes caused massive apoptosis in these cells. Consequently, localized expression of the gene in tumors may be a good way to reduce systemic toxic effects while providing a constant therapeutic effect at the cancer site. The goal of this proposal is to develop technology that can target pharmaceutical effects of the TRAIL gene to cancer cells. We hypothesize that the promoter for human telomerase reverse transcriptase (hTERT), active in 85 percent of primary human cancers, will be able to elicit tumor-specific expression of the TRAIL gene and thus can be used to avoid toxicity in susceptible normal cells. In a preliminary study, we found that in vitro transfer of the TRAIL gene elicited apoptosis and bystander effects in colon cancer DLD-1 cells but not in normal fibroblasts. Intratumoral delivery of the TRAIL gene effectively suppressed tumor growth of DLD-1 xenograft. We also demonstrated that the hTERT promoter could induce high levels of transgene expression in cancer cells but not in normal cells in vitro and in vivo. Our preliminary results showed that the hTERT promoter can be used to prevent GFP-TRAIL fusion gene expression in liver cells in vitro and in vivo. To achieve our goals, we will assess therapeutic value of the TRAIL gene expressed from the hTERT promoter. In vitro and in vivo studies are proposed to evaluate the efficiency of gene transfer, levels of transgene expression, apoptosis induction, the bystander effect and its possible mechanism, antitumor activity, anti-metastasis activity and possible toxicity of the GFP/TRAIL gene expressed from the hTERT promoter and delivered by adenoviral vector. Completion of the proposed studies will allow us to develop technologies for targeting the pharmaceutical effects of the TRAIL gene to cancers so that the treatment will have maximum therapeutic effects but minimum toxicity. The preclinical documentation we obtain on tumor-specific expression driven by the hTERT promoter and on the therapeutic/toxic profiles of hTERT-TRAIL constructs may provide a scientific basis for future clinical application of this approach in humans.

由于它们在多种肿瘤细胞中引起细胞死亡的能力，但在正常细胞中却没有，因此与TNFalpha相关 - 凋亡诱导的配体（TRAIL）的重组蛋白（TRAIL）似乎是癌症患者的有前途药物。然而，最近在正常人肝细胞中，重组径径蛋白凋亡诱导的凋亡诱导的发现引起了人们对当系统施用TRAIL蛋白时严重毒性的可能性的警报。 我们的初步研究还表明，在原代人肝细胞中，全长径径蛋白的表达在这些细胞中引起大量凋亡。 因此，肿瘤中基因的局部表达可能是减少全身毒性作用的好方法，同时在癌症部位提供持续的治疗作用。 该提案的目的是开发可以靶向TRAIL基因对癌细胞的药物作用的技术。 我们假设人类端粒酶逆转录酶（HTERT）的启动子在85％的原发性人类癌症中活跃，将能够引起TRAIL基因的肿瘤特异性表达，因此可以用于避免在敏感正常细胞中的毒性。 在一项初步研究中，我们发现，在结肠癌DLD-1细胞中，TRAIL基因的体外转移引起了凋亡和旁观者的作用，但在正常成纤维细胞中却没有。 TRAIL基因的肿瘤内递送有效地抑制了DLD-1异种移植的肿瘤生长。 我们还证明了HTERT启动子可以在癌细胞中诱导高水平的转基因表达，而在正常细胞中不能在体外和体内诱导。 我们的初步结果表明，HTERT启动子可用于预防体外和体内肝细胞中的GFP-Trail融合基因表达。 为了实现我们的目标，我们将评估从HTERT启动子表达的TRAIL基因的治疗价值。 提出了体外和体内研究，以评估基因转移水平，转基因表达水平，凋亡诱导水平，旁观者效应及其可能的机制，抗肿瘤活性，抗肿瘤活性以及GFP/TRAIL基因可能从HTERT启动子表达的GFP/TRAIL基因的毒性。 拟议的研究的完成将使我们能够开发针对癌症对癌症的药物作用的技术，从而使治疗具有最大的治疗作用，但最小的毒性。 我们在HTERT启动子驱动的肿瘤特异性表达以及HTERT-TRAIL构建体的治疗/有毒谱上获得的临床前文献可能为这种方法在人类中的未来临床应用提供了科学基础。