锌指核酶靶向治疗CIN及宫颈癌的实验研究

项目名称： 锌指核酶靶向治疗CIN及宫颈癌的实验研究

项目编号： No.81502252

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 丁文成

作者单位： 华中科技大学

项目金额： 19万元

中文摘要： 高危型人乳头状瘤病毒（HPV）特别是16、18亚型HPV的感染是宫颈癌发生发展的重要病理因素，靶向HPV的基因治疗具有重大的理论意义和临床应用价值，然而目前尚没有高效且特异性的靶向HPV的基因治疗手段出现。本课题组前期通过引进高效的基因修饰技术锌指核酶（ZFNs），通过系统的的体外细胞实验，证实其能高效且特异性的诱导相应HPV亚型阳性细胞的凋亡和阻滞细胞的增殖，而对其他亚型阳性及HPV阴性细胞均无作用。本项目拟延续前期的研究成果，通过在小鼠阴道内直接转染EGFP或mRFP荧光蛋白质粒来建立和优化阴道内转染技术，通过在系统表达EGFP转基因小鼠阴道内转染靶向EGFP的ZFNs，明确ZFNs局部应用的效率，在K14-HPV16转基因小鼠（CIN模型）阴道内转染靶向HPV16 E6E7基因的ZFNs，在动物模型内探索利用ZFNs靶向防治CIN和宫颈癌的可行性。

中文关键词： 子宫颈肿瘤；基因治疗；锌指核酶；人乳头状瘤病毒；靶向治疗

英文摘要： Infection of high-risk human papillomavirus (HPV), especially HPV16 and HPV18, is a vital pathological factor of the development of cervical cancer. Gene therapy of targeting HPV has a potential value of clinical application. However, there have not exist any approach with high efficiency and specificity. Our group had introduced zinc finger nucleases (ZFNs), which is a high efficient gene editing technology, into research of gene therapy. And our previous experiments has confirmed that ZFNs can induce cell apoptosis and inhibit proliferation of corresponding HPV positive cells, while cannot affect other HPV positive or HPV negative cells. This program is a continuation of the previous research. We plan to establish and optimize intravaginal transfection through transfecting EGFP or MRFP expression plasmids into mice vagina directly. Then determine the efficiency of topical transfection of ZFNs through transfection of ZFNs targeting EGFP into systemic expression EGFP transgenic mice. At last, using topical transfection of ZFNs targeting HPV16 E6E7 into K14-HPV16 transgenic mice (a CIN model) to explore the feasibility of ZFNs in prevention and treatment of CIN even cervical cancer.

英文关键词： Cervical Cancer;Gene Therapy;Zinc Finger Nuclease;Human Papillomavirus;Target Therapy