Molecular pathogenesis and host response following persistent E6/E7 expression

E6/E7 持续表达后的分子发病机制和宿主反应

• 批准号: 8619069
• 负责人: Chien-Fu Hung
• 金额: $ 20.25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-08 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8619069
• 关键词: Animal Model; Anogenital Human Papilloma Virus Infection; Antigens; Antiviral Agents; Anus; Anxiety; Biological; Biological Process; C57BL/6 Mouse; Cancerous; Cervix Uteri; Chronic; DNA; Data; Development; Disease; Event; Frequencies; Generations; Genes; Genital Human Papilloma Virus Infection; HPV-High Risk; Histopathology; Human Papilloma Virus Vaccine; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Immune; Immune response; Immunocompetent; Immunocompromised Host; Immunosuppression; Individual; Infection; Intervention; Intraepithelial Neoplasia; Kinetics; Lesion; Luciferases; Malignant - descriptor; Malignant Neoplasms; Malignant Vaginal Neoplasm; Malignant neoplasm of anus; Malignant neoplasm of vulva; Mediating; Methodology; Modeling; Molecular; Morbidity - disease rate; Mus; Oncogenes; Oncogenic; Operative Surgical Procedures; Pathogenesis; Patients; Pharmaceutical Preparations; Pre-Clinical Model; Premalignant; Proteins; Public Health; Recurrence; Research; Risk; Severities; Sexually Transmitted Diseases; Staging; System; T-Lymphocyte; Testing; Therapeutic; Vaccination; Vagina; Vulva; adaptive immunity; base; carcinogenesis; cell mediated immune response; effective therapy; immunosuppressed; innovation; insight; intraepithelial; non-invasive imaging; novel; public health relevance; ras Oncogene; screening; young man; young woman

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.

项目概要/摘要 人乳头瘤病毒（HPV）是最常见的性传播感染。持续感染 HPV 是 流行于年轻男性和女性，感染高危 HPV 类型，尤其是 HPV-16，与 发生上皮内瘤变的风险增加。此外，免疫抑制的个体 感染 HPV 并随后发生与 HPV 相关的癌前病变的风险更大，并且 癌症病变。目前，尚无针对持续性生殖器 HPV 感染的抗病毒治疗方法。目前的 对此类患者的管理仅包括重复筛查癌前病变和癌症 病变，从而产生明显的不适和焦虑。此外，90% 的 HPV 相关阴道、 外阴癌和肛门癌可归因于 HPV-16。此外，虽然手术治疗对于 宫颈癌前病变和早期癌病变，与显着的发病率和高复发率相关 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 治疗以消除阴道、外阴和肛门中持续存在的高危 HPV 感染，尤其是 HPV-16。在 为了开发针对持续性 HPV-16 感染和 HPV-16 相关肛门生殖器的有效治疗方法 病变，开发慢性 HPV 感染的临床前模型至关重要。现在已经明确的是，高危型HPV E6和E7致癌蛋白负责HPV相关病变的恶性进展。我们 最近生成了携带能够表达荧光素酶的 DNA 构建体的 HPV-16 假病毒体 (psV)， HPV E6 和 E7 癌基因以及 Ras 癌基因分别。我们发现小鼠感染了 HPV-16 psV 阴道感染后 150 天持续表达荧光素酶以及 E6 和 E7 癌基因。在 相比之下，携带荧光素酶对照 DNA 的 HPV-16 psV 仅表现出瞬时表达 编码基因。因此，我们创建了一个非侵入性成像系统来跟踪持续表达 E6/E7 在阴道中。当前项目的目的是表征动物模型是否类似于 持续性肛门生殖器 HPV 感染并研究其分子发病机制和免疫反应 持续感染进展为癌前病变和癌症病变。我们的具体目标是：（1）描述 HPV-16 E6/E7和Ras在阴道、外阴和肛门持续表达的分子发病机制 免疫功能健全和免疫功能低下的宿主和（2）表征E6和E7特异性体液和细胞- HPV-16 pVs 持续肛门生殖器感染后介导的免疫反应。成功者 该项目的实施将产生持续表达 HPV-16 E6/E7 的临床前模型， 类似于慢性高危HPV感染。这将代表一种真正创新的方法，可能会改变 HPV 研究的典范。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 致癌蛋白的干预措施，用于控制 HPV 相关疾病。结果 这项研究将对持续性 HPV 感染的管理和治疗产生重大影响。