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.

项目总结/摘要 人乳头瘤病毒（HPV）是最常见的性传播感染。持续性HPV感染 在年轻男性和女性中流行，感染高危型HPV，特别是HPV-16， 增加了发展上皮内瘤变的风险。此外，免疫抑制的个体 有更大的风险感染HPV，随后发展为HPV相关的癌前病变， 癌症病变。目前，没有针对持续生殖器HPV感染的抗病毒治疗。当前 对这类患者的管理仅包括重复筛查癌前病变和癌症的发展 病变，这会产生显著的不适和焦虑。此外，90%的HPV相关阴道， 外阴和肛门癌可归因于HPV-16。此外，虽然手术治疗对 宫颈癌前病变和早期癌病变，与显著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、外阴和肛门中持续的高危HPV感染，特别是HPV-16。在 为了开发一种有效的治疗持续性HPV-16感染和HPV-16相关的肛门生殖器感染的方法， 病变，开发慢性HPV感染的临床前模型至关重要。现在很清楚，高危HPV E6和E7致癌蛋白负责HPV相关病变的恶性进展。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）， HPV E6、E7癌基因和Ras癌基因。我们发现感染了HPV-16 psV的小鼠 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，携带对照DNA和荧光素酶的HPV-16 psV仅表现出瞬时表达。 编码基因因此，我们已经创建了一个非侵入性成像系统，以跟踪 E6/E7在阴道内。目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，并研究分子发病机制和免疫反应， 持续性感染进展为癌前病变和癌病变。我们的具体目标是：（1）表征 HPV-16 E6/E7和Ras在女性外阴、阴道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特异性体液和细胞- HPV-16 psV持续性肛门生殖器感染后介导的免疫应答。成功 该项目实施将产生持续HPV-16 E6/E7表达的临床前模型， 类似于慢性高危HPV感染。这将代表一个真正创新的方法论，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，以控制HPV相关疾病。的结果 这项研究将对持续性HPV感染的管理和治疗具有重要意义。