TALEN-mediated targeting of HPV oncogenes ameliorates HPV-related cervical malignancy

• Published in: The Journal of clinical investigation Vol. 125; no. 1; pp. 425 - 436
• Main Authors: Hu, Zheng, Ding, Wencheng, Zhu, Da, Yu, Lan, Jiang, Xiaohui, Wang, Xiaoli, Zhang, Changlin, Wang, Liming, Ji, Teng, Liu, Dan, He, Dan, Xia, Xi, Zhu, Tao, Wei, Juncheng, Wu, Peng, Wang, Changyu, Xi, Ling, Gao, Qinglei, Chen, Gang, Liu, Rong, Li, Kezhen, Li, Shuang, Wang, Shixuan, Zhou, Jianfeng, Ma, Ding, Wang, Hui
• Format: Journal Article
• Language: English
• Published: United States American Society for Clinical Investigation 01.01.2015
• Subjects: Animals; Antineoplastic Agents - therapeutic use; Apoptosis; Biomedical research; Cell Cycle Checkpoints; Cervical cancer; Complications and side effects; Deoxyribonucleases - therapeutic use; DNA Cleavage; DNA, Viral - genetics; Female; Genetic aspects; Health aspects; HEK293 Cells; HeLa Cells; Human papillomavirus; Human papillomavirus 16 - genetics; Humans; Medical research; Mice, Inbred BALB C; Mice, Nude; Mice, Transgenic; Mutation; Oncogene Proteins, Viral - genetics; Papillomavirus E7 Proteins - genetics; Papillomavirus infections; Papillomavirus Infections - therapy; Papillomavirus Infections - virology; Physiological aspects; Plasmids; Repressor Proteins - genetics; Risk factors; Studies; Tumor proteins; Uterine Cervical Neoplasms - therapy; Uterine Cervical Neoplasms - virology; Xenograft Model Antitumor Assays; China
• ISSN: 0021-9738; 1558-8238; 1558-8238
• DOI: 10.1172/JCI78206

Persistent HPV infection is recognized as the main etiologic factor for cervical cancer. HPV expresses the oncoproteins E6 and E7, both of which play key roles in maintaining viral infection and promoting carcinogenesis. While siRNA-mediated targeting of E6 and E7 transcripts temporarily induces apoptosis in HPV-positive cells, it does not eliminate viral DNA within the host genome, which can harbor escape mutants. Here, we demonstrated that specifically targeting E6 and E7 within host DNA with transcription activator-like effector nucleases (TALENs) induces apoptosis, inhibits growth, and reduces tumorigenicity in HPV-positive cell lines. TALEN treatment efficiently disrupted E6 and E7 oncogenes, leading to the restoration of host tumor suppressors p53 and retinoblastoma 1 (RB1), which are targeted by E6 and E7, respectively. In the K14-HPV16 transgenic mouse model of HPV-driven neoplasms, direct cervical application of HPV16-E7-targeted TALENs effectively mutated the E7 oncogene, reduced viral DNA load, and restored RB1 function and downstream targets transcription factor E2F1 and cycling-dependent kinase 2 (CDK2), thereby reversing the malignant phenotype. Together, the results from our study suggest that TALENs have potential as a therapeutic strategy for HPV infection and related cervical malignancy.