TALEN-mediated targeting of HPV oncogenes ameliorates HPV-related cervical malignancy
TALEN-mediated targeting of HPV oncogenes ameliorates HPV-related cervical malignancy
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TALEN 介导的 HPV 癌基因靶向可改善 HPV 相关宫颈恶性肿瘤
DOI：
10.1172/jci78206
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发表时间：
2015-01-01
影响因子：
15.9
通讯作者：
Wang, Hui
中科院分区：
文献类型：
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作者：
Hu, Zheng;Ding, Wencheng;Wang, Hui
持续性人乳头瘤病毒（HPV）感染被认为是宫颈癌的主要病因。HPV表达癌蛋白E6和E7，这两种蛋白在维持病毒感染和促进癌变过程中都起着关键作用。虽然小干扰RNA（siRNA）介导的对E6和E7转录本的靶向作用可暂时诱导HPV阳性细胞凋亡，但它不能消除宿主基因组内的病毒DNA，而这些病毒DNA可能含有逃逸突变体。在此，我们证明了利用转录激活因子样效应物核酸酶（TALENs）特异性靶向宿主DNA中的E6和E7可诱导凋亡、抑制生长，并降低HPV阳性细胞系的致瘤性。TALEN治疗有效地破坏了E6和E7癌基因，导致宿主肿瘤抑制因子p53和视网膜母细胞瘤1（RB1）的功能恢复，E6和E7分别作用于这两种因子。在HPV驱动的肿瘤的K14 - HPV16转基因小鼠模型中，直接在宫颈部位应用靶向HPV16 - E7的TALENs可有效使E7癌基因发生突变，降低病毒DNA载量，恢复RB1功能以及下游靶点转录因子E2F1和细胞周期蛋白依赖性激酶2（CDK2）的功能，从而逆转恶性表型。总之，我们的研究结果表明，TALENs有作为HPV感染及相关宫颈恶性肿瘤治疗策略的潜力。
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.