Molecular pathogenesis and host response following persistent E6/E7 expression

项目摘要/摘要 人乳头瘤病毒(HPV)是最常见的性传播感染。持续的HPV感染是 流行于年轻男性和女性，感染高危HPV类型，特别是HPV-16，与 发生上皮内瘤变的风险增加。此外，免疫抑制的个体 有更大的风险感染HPV并随后发展为HPV相关的癌前病变和 癌症损伤。目前，还没有针对持续性生殖器HPV感染的抗病毒治疗。海流 对这类患者的管理只包括对癌症前期和癌症的重复筛查。 损伤，这会产生严重的不适和焦虑。此外，在所有与HPV相关的阴道中， 外阴癌和肛门癌可归因于HPV-16。此外，虽然手术治疗对 宫颈癌前病变和早期癌病变，发病率高，复发率高。 阴道、外阴和肛门上皮内病变的发生率。因此，迫切需要开发一种创新的 治疗以消除持续的高危HPV感染，特别是阴道、外阴和肛门的HPV-16感染。在……里面 为开发治疗HPV-16持续感染和HPV-16相关生殖器的有效方法 因此，有必要建立一种慢性HPV感染的临床前模型。现在很明显，高危HPV E6和E7癌基因蛋白与HPV相关病变的恶性进展有关。我们 最近产生了携带能够表达荧光素酶的DNA构建体的HPV-16假病毒粒子(PSV)， HPVE6、E7癌基因和RAS癌基因。我们发现感染这种HPV-16PSV的小鼠 阴道感染150天后，荧光素酶及E6、E7癌基因持续表达。在……里面 相比之下，携带对照DNA和荧光素酶的HPV-16PSV仅显示瞬时表达 编码的基因。因此，我们创建了一种非侵入性成像系统，以遵循持续表达的 E6/E7在阴道中。当前项目的目的是确定动物模型是否类似于 目的：探讨HPV持续感染的分子发病机制和免疫应答。 持续感染会进展为癌前病变和癌症损害。我们的具体目标是：(1)描述 HPV-16E6/E7和RAS在人阴道、外阴和肛门持续表达的分子发病机制 免疫活性和免疫受损宿主和(2)表征E6和E7特异的体液和细胞- HPV-16PSV持续肛门感染后介导的免疫反应。成功者 该项目的实施将产生HPV-16E6/E7持续表达的临床前模型， 类似于慢性高危HPV感染。这将代表一种可能会改变的真正创新的方法 人乳头瘤病毒研究中的范式。此外，我们的临床前模型将促进分子生物学的发展。 针对E6/E7癌基因蛋白的干预措施，用于控制HPV相关疾病。结果是 本研究对持续HPV感染的管理和治疗具有重要意义。

PROJECT SUMMARY/ABSTRACT Human papillomavirus (HPV) is the most common sexually transmitted infection. Persistent HPV infection is prevalent in young men and women and infection with a high-risk HPV type, especially HPV-16, is associated with an increased risk of developing an intraepithelial neoplasia. Furthermore, immunosuppressed individuals have a greater risk of being infected with HPV and subsequently developing HPV-associated precancer and cancer lesions. Currently, there is no antiviral treatment for persistent genital HPV infection. The current management for such patients only includes repeat screening for the development precancer and cancer lesions, which generates significant discomfort and anxiety. Furthermore, 90% of all HPV-associated vaginal, vulval and anal cancers are attributable to HPV-16. Additionally, while surgical treatment is quite effective for precancer and early cancer lesions of the cervix, it is associated with significant morbidity and high recurrence rates in vaginal, vulval and anal intraepithelial lesions. Thus, there is an urgent need to develop an innovative treatment to eliminate persistent, high-risk HPV infections, especially HPV-16, in the vagina, vulva and anus. In order to develop an effective treatment for persistent HPV-16 infection and HPV-16-associated anogenital lesions, it is essential to develop a preclinical model of chronic HPV infection. It is now clear that high-risk HPV E6 and E7 oncogenic proteins are responsible for the malignant progression of HPV-associated lesions. We recently generated an HPV-16 pseudovirion (psV) carrying a DNA construct capable of expressing luciferase, HPV E6 and E7 oncogenes and Ras oncogene individually. We found that mice infected with this HPV-16 psV persistently expressed luciferase as well as E6 and E7 oncogenes 150 days after infection of the vagina. In comparison, HPV-16 psV carrying control DNA with luciferase only demonstrated transient expression of the encoded gene. Thus, we have created a non-invasive imaging system to follow the persistent expression of E6/E7 in the vagina. The purpose of the current project is to characterize whether the animal model resembles persistent anogenital HPV infection and to investigate the molecular pathogenesis and immune responses as persistent infection progresses to precancer and cancer lesions. Our specific aims are to: (1) characterize the molecular pathogenesis of persistent HPV-16 E6/E7 and Ras expression in the vagina, vulva and anus of immunocompetent and immunocompromised hosts and (2) characterize E6 and E7-specific humoral and cell- mediated immune responses following persistent anogenital infection with HPV-16 psVs. The successful implementation of this project will generate a preclinical model of persistent HPV-16 E6/E7 expression, resembling chronic high risk HPV infection. This will represent a truly innovative methodology that may change the paradigm in HPV research. Additionally, our preclinical model will facilitate the development of molecular interventions targeting E6/E7 oncogenic proteins for the control of HPV-associated diseases. The results of this study will have significant implications for the management and treatment of persistent HPV infection.