A Humanized Mouse Model of HPV-Associated Pathology Driven by E7 Expression
A Humanized Mouse Model of HPV-Associated Pathology Driven by E7 Expression
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DOI：
10.1371/journal.pone.0041743
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发表时间：
2012-07-23
期刊：
影响因子：
3.7
通讯作者：
Garcia-Escudero, Ramon
中科院分区：
文献类型：
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作者：
Buitrago-Perez, Agueda;Hachimi, Mariam;Garcia-Escudero, Ramon
人乳头瘤病毒（HPV）是人类宫颈癌的病原体，并且与口咽鳞状细胞癌的发生有关。尽管已经研发出预防性疫苗，但仍需要为这些部位患有恶性感染性病变的个体开发新的靶向疗法，且必须在合适的模型中进行测试。皮肤β型HPV似乎与皮肤癌发生有关。病毒的致癌性部分是通过病毒E7基因使视网膜母细胞瘤蛋白家族成员失活来实现的。在此我们表明，皮肤β型HPV5的E7蛋白与pRb结合并促进其降解。此外，我们描述了一种HPV相关疾病的体内模型，在该模型中，使用经过基因工程改造以表达E7蛋白的原代角质形成细胞制备的人造皮肤被移植到裸鼠身上。移植体中E7的表达可稳定维持长达6个月，诱导病变出现，就HPV16 E7而言，其在组织学上类似于致癌性HPV引起的人类肛门生殖器病变。此外，通过免疫检测和/或从mRNA和miRNA进行定量PCR对生物标志物表达进行分析证实，16E7修饰的移植皮肤与人类HPV相关的肿瘤前和肿瘤病变具有分子特征。最后，我们的研究结果表明，HPV5 E7在体内降低pRb水平的体外能力下降，这可能解释了与16E7移植体相比的表型差异。我们的模型似乎是HPV致癌作用基础研究以及HPV相关抗肿瘤疗法临床前测试的一个有价值的平台。
Human papillomavirus (HPV) is the causative agent of human cervical cancer and has been associated with oropharyngeal squamous cell carcinoma development. Although prophylactic vaccines have been developed, there is a need to develop new targeted therapies for individuals affected with malignant infected lesions in these locations, which must be tested in appropriate models. Cutaneous beta HPV types appear to be involved in skin carcinogenesis. Virus oncogenicity is partly achieved by inactivation of retinoblastoma protein family members by the viral E7 gene. Here we show that the E7 protein of cutaneous beta HPV5 binds pRb and promotes its degradation. In addition, we described an in vivo model of HPV-associated disease in which artificial human skin prepared using primary keratinocytes engineered to express the E7 protein is engrafted onto nude mice. Expression of E7 in the transplants was stably maintained for up to 6 months, inducing the appearance of lesions that, in the case of HPV16 E7, histologically resembled human anogenital lesions caused by oncogenic HPVs. Moreover, it was confirmed through biomarker expression analysis via immunodetection and/or quantitative PCR from mRNA and miRNA that the 16E7-modified engrafted skin shares molecular features with human HPV-associated pretumoral and tumoral lesions. Finally, our findings indicate a decrease of the in vitro capacity of HPV5 E7 to reduce pRb levels in vivo, possibly explaining the phenotypical differences when compared with 16E7-grafts. Our model seems to be a valuable platform for basic research into HPV oncogenesis and preclinical testing of HPV-associated antitumor therapies.