Project 2-Three-dimensional modeling of EBV-HPV interactions leading to anogenital cell dysplasia

项目 2-EBV-HPV 相互作用导致肛门生殖器细胞发育不良的三维模型

• 批准号: 9209611
• 负责人: Chris L McGowin
• 金额: $ 20.1万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9209611
• 关键词: 3-Dimensional; Address; Affect; African American; Architecture; Area; Basal Cell; Basement membrane; Biopsy; Cancer cell line; Cell Proliferation; Cells; Centers of Research Excellence; Cervical; Cervical dysplasia; Cervix Uteri; Cervix carcinoma; Clinical; Collaborations; Consult; Data; Detection; Development; Disease; Dysplasia; Environment; Epithelial; Epithelial Cells; Epithelium; Epstein-Barr Virus Infections; Funding; Future; Genomic DNA; Goals; HIV; HPV-High Risk; High Risk Woman; Human; Human Herpesvirus 4; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Human papillomavirus 18; Human papillomavirus 6; Imagery; Immunohistochemistry; Infection; Invaded; Investigation; Laboratories; Leadership; Life Cycle Stages; Link; Louisiana; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Measures; Mentors; Methods; Microscopy; Modeling; Molecular Target; Oncogenes; Oncogenic; Oncogenic Viruses; Oncoproteins; Outcome; Outcome Measure; Papillomavirus; Pathogenicity; Phenotype; Production; Property; Research Personnel; Retroviridae; Risk; Sampling; Scientist; Sexually Transmitted Diseases; Squamous intraepithelial lesion; Staining method; Stains; System; Tissues; Training; Translating; Viral; Virus; Virus Diseases; Woman; carcinogenesis; carcinogenicity; career; co-infection; cohort; design; experimental study; field study; high risk; innovation; insight; keratinocyte; keratinocyte differentiation; member; metaplastic cell transformation; monolayer; novel; oncology; penis foreskin; retroviral transduction; three-dimensional modeling; tumorigenesis; two-dimensional

ABSTRACT Infections with high-risk sub-types of Human Papilloma Virus (HPV) are responsible for more than 99% of all cervical carcinoma cases in the US. Our recent data in collaboration with Dr. Hagensee laboratory has shown that co-detection of Epstein-Barr virus (EBV) with HPV in cervical samples increases the risk of concurrent cervical dysplasia by 4-6 fold. In addition, HIV+ women from New Orleans (n=531) with detectable cervical HPV and EBV are at higher risk (69%) for concurrent squamous intraepithelial lesions (SIL) as compared to only 28% of women with only detectable HPV (p<0.001; OR 5.57, 95%CI 2.19-14.4). We hypothesize that EBV acts as a co-factor for progression of HPV-related cervical cancers. We will address this hypothesis directly using innovative 3-dimensional (3D) epithelial models of co-infection. First, using a well-characterized 3D model of terminally differentiated keratinocytes, we will determine whether co-infection with HPV and EBV adversely affects normal differentiation of the stratified multi-layer epithelium compared to HPV alone. Using targeted molecular methods, we will dissect the augmenting capacity of EBV's oncoprotein, LMP-1, for cellular transformation in the context of high-risk HPV types. Importantly, the proposed outcome measures will also address epithelial transformation in the context of the low-risk HPV6 because HPV6 is found more frequently in women shedding EBV, and since EBV's augmenting capacity might be greater in otherwise low-risk types. Parallel experiments will be performed in primary and/or HPV-immortalized human ectocervical epithelial cells. Utilizing expertise from all members of the COBRE team, the innovative 3D epithelial models utilized herein will facilitate a powerful and direct investigation of whether co-infection with EBV enhances malignant transformation. Specifically, these models allow for visualization of the stratified epithelial architecture and invasion of the underlying stromal layer, quantification of basal cell proliferation, and several outcomes not achievable with conventional 2D (monolayer) models. Also unique to the 3D model, we have the ability to produce and utilize infectious HPV thus allowing for co-infection with live viruses rather than modeling HPV infection by retroviral transduction of E6/E7 oncogenes. The mentoring duo of Drs. Alison Quayle and Augusto Ochoa, combined with consulting provided by Drs. Michelle Ozbun and Hagensee, will provide excellent scientific leadership to proposed COBRE mentee Dr. Chris McGowin. In line with the goals of the COBRE mechanism, the combined track record of the proposed mentors for training of junior investigators will provide Chris a rich environment conducive for translating his findings into independent R01 funding.

摘要 人类乳头瘤病毒（HPV）的高风险亚型感染占所有病例的99%以上。 美国宫颈癌病例。我们与哈根西博士实验室合作的最新数据显示， 宫颈样本中EB病毒（EBV）与HPV的共同检测增加了同时发生 宫颈发育不良4-6倍。此外，来自新奥尔良的HIV+妇女（n=531）， HPV和EBV并发鳞状上皮内病变（SIL）的风险较高（69%）， 仅28%的女性仅检测到HPV（p<0.001; OR 5.57，95%CI 2.19-14.4）。我们假设EB病毒 作为HPV相关宫颈癌进展的辅助因素。我们将直接讨论这个假设 使用创新的三维（3D）上皮模型的共同感染。首先，使用特征良好的3D 终末分化的角质形成细胞模型，我们将确定是否与HPV和EBV共感染 与单独的HPV相比，其不利地影响复层多层上皮的正常分化。使用 靶向分子方法，我们将剖析EB病毒的癌蛋白，LMP-1，细胞增殖的能力， 在高危HPV类型的背景下转化。重要的是，拟议的成果措施还将 在低风险HPV 6的背景下解决上皮转化，因为HPV 6在 妇女脱落EBV，因为EBV的扩增能力可能更大，否则低风险类型。 平行实验将在原代和/或HPV永生化人宫颈上皮细胞中进行。 利用COBRE团队所有成员的专业知识，本文所用的创新3D上皮模型将 有助于强有力的和直接的调查是否与EBV的共同感染增强恶性肿瘤的发生。 转型具体地，这些模型允许分层上皮结构的可视化， 下面的基质层的侵袭，基底细胞增殖的定量，以及一些结果， 可通过常规2D（单层）模型实现。3D模型也是独一无二的，我们有能力 产生和利用感染性HPV，从而允许与活病毒共感染，而不是模拟HPV 通过逆转录病毒转导E6/E7癌基因感染。艾莉森·奎尔博士和奥古斯托博士 奥乔亚，结合米歇尔·奥兹本博士和哈根西博士提供的咨询，将提供出色的 科学领导力，以建议COBRE学员克里斯·麦高文博士。根据COBRE的目标， 根据这一机制，拟议的初级调查员培训导师的综合业绩记录将提供 克里斯有一个丰富的环境，有利于将他的发现转化为独立的R 01资金。