Anti-HPV RNA Interference Using Modified RNA's

使用修饰 RNA 进行抗 HPV RNA 干扰

• 批准号: 6935803
• 负责人: KENNETH A ALEXANDER
• 金额: $ 7.58万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-11 至 2005-11-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6935803
• 关键词: HeLa cells; RNA; RNA interference; antisense nucleic acid; bioengineering /biomedical engineering; biotechnology; cancer prevention; cell senescence; chemical stability; chemical structure function; chemical synthesis; gene induction /repression; green fluorescent proteins; human papillomavirus; neoplasm /cancer therapy; nucleic acid structure; preneoplastic state; radiotracer; therapy design /development; thiophosphate; tissue /cell culture; virus diseases; virus genetics; virus protein; virus related neoplasm /cancer

DESCRIPTION (provided by applicant): Human papillomaviruses (HPVs) are the major cause of anogenital dysplasias and carcinomas in both men and women. The HPV E6 and E7 proteins underlie the ability of papillomaviruses to cause disease. We have used short interfering double strand RNAs directed against HPV-18 E6 and E7 (RNA interference) to successfully cause growth arrest and senescence in HeLa cells. Similar results have been obtained in SW756 cells (a second HPV-18-transformed cell line), but not in C33A cells (a non-HPV-transformed cervical carcinoma cell line). Our data indicate that anti-E6/E7 RNA interference can reduce E6 and E7 expression, and that the reduction of E6 and E7 expression could be used to treat both pre-malignant HPV infections and HPV-related cancers. A barrier to the therapeutic use of RNA interference is the inefficiency with which double strand phosphodiester RNAs enter cells. To promote the entry of therapeutic interfering RNAs into cells without the use of lipofection reagents, we have begun to evaluate boranophosphate- and phosphorothioate-backbone RNAs for their abilities to enter cells and to induce RNA interference. Our data indicate that both boranophosphate- and phosphorothioate-backbone RNAs can be used to induce RNA interference of green fluorescent protein expression in HeLa cells. Boranophosphate-modified single-stranded siRNAs are also active. We predict that boranophosphate and phosphorothioate RNAs will enter cells efficiently, and will induce RNA interference with E6 and E7 expression, resulting in growth arrest and senescence of HeLa cells in culture. RNA interference holds significant early promise for the treatment of many disorders, including HPV infections and HPV-related malignancies. This work is an initial step in the evaluation of boranophosphateand phosphorothioate-based RNA interference as a therapeutic modality.

描述（由申请人提供）：人乳头瘤病毒（hpv）是男性和女性生殖器发育不良和癌的主要原因。HPV E6和E7蛋白是乳头瘤病毒致病能力的基础。我们使用短干扰双链RNA靶向HPV-18 E6和E7 （RNA干扰），成功地引起HeLa细胞的生长停滞和衰老。在SW756细胞（第二种hpv -18转化细胞系）中获得了类似的结果，但在C33A细胞（一种非hpv转化的宫颈癌细胞系）中没有得到类似的结果。我们的数据表明，抗E6/E7 RNA干扰可以降低E6和E7的表达，并且降低E6和E7的表达可以用于治疗恶性前HPV感染和HPV相关癌症。