The role of Epstein Barr Virus and Human Papilloma Virus Co-infection in Conjunctival Squamous Cell Carcinoma.

项目总结 结膜鳞状细胞癌是眼部的一种恶性疾病，归类于眼科 表面鳞状肿瘤(OSSN)(Gichuhi等人)。2015年)。结膜鳞状细胞癌的发病率有地理分布 多变性，发病率高发生在非洲(每10万人口每年约1.3人)，而 这在世界其他地区相对罕见(每10万人每年约有0.1人)(吉竹市和 Sagoo 2016；Gichuhi等人。2013)。我们建议，第一次评估传染病病原体的作用， 人乳头瘤病毒(HPV)和EB病毒(EBV)在津巴布韦结膜鳞状细胞癌中的感染。这将建立 根据我们最近的初步数据(Galati等人，2020年)，其中爱泼斯坦-巴尔病毒(EBV)感染率高， 来自乌干达结膜鳞状细胞癌患者组织中61%的人乳头瘤病毒(HPV)是 检测到。 目前拟议的研究将涉及三个目标。 1.比较组织学证实的组织样本中是否存在乙型HPV和EBV 结膜鳞状细胞癌患者(病例)与非恶性病变的非肿瘤对照 从津巴布韦哈拉雷的一家三级医院和两家私人实验室招募。 2.确定病毒致癌蛋白表达的流行率(例如，HPV的E6和EBV的LMP1) 在结膜鳞癌组织中， 3.探索EBV转录本在不同细胞类型(B细胞、T细胞和上皮细胞)中的位置 在结膜鳞状细胞癌FFPE组织样本中，按HPV合并感染和HIV状态分层。 研究设计将是一项病例对照的回溯性研究，检索估计200例连续的癌症 样本(病例)和200例非肿瘤对照。病例和对照将按性别、年龄(+/-10岁)匹配 在可能的情况下，2015年1月1日至31日期间经组织学确诊的艾滋病毒状况 2020年1月，来自津巴布韦哈拉雷的中央第三级组织病理学实验室，那里的大多数省份 私立医院将他们的样本提交给。病毒DNA和RNA将在上皮和/或B和T细胞中检测到 通过基于聚合酶链式反应和原位杂交的方法从FFPE样本中提取细胞。我们的学习将建立在我们强大的 初步数据，并预计将提供证据，证明参与的独特组合 结膜鳞状细胞癌的感染性病原体(EBV/HPV)我们的发现将为预防和预防 筛查针对非洲人群的干预措施，在非洲，这种恶性肿瘤更常见。

PROJECT SUMMARY Conjunctival Squamous Cell Carcinoma (SCC) is a malignant disease of the eyes, classified under ocular surface squamous neoplasia (OSSN) (Gichuhi et al. 2015). Incidence of conjunctival SCC has a geographic variability, with high incidence rates occurring in Africa (about 1.3 people per 100,000 population- year), whilst it is relatively rare in other regions of the world (about 0.1 people per 100,000 population-year) (Gichuhi and Sagoo 2016; Gichuhi et al. 2013). We propose to evaluate, for the first time, the role of infectious agents, Human Papilloma Virus (HPV) and Epstein Barr Virus (EBV) in conjunctival SCC in Zimbabwe. This will build upon our recent preliminary data (Galati et al. 2020), where high rates of both Epstein-Barr virus (EBV), 64% and beta human papillomavirus (HPV), 61% in tissue from Ugandan patients with conjunctival SCC, were detected. Three aims will be addressed in the current proposed research. 1. Compare the presence of beta HPV and EBV in tissue samples from histologically confirmed conjunctival SCC patients (cases) versus cancer-free controls with non-malignant pathologies recruited from a tertiary hospital and two private laboratories in Harare, Zimbabwe. 2. Determine the prevalence of viral oncogenic protein expression (e.g., E6 for HPV and LMP1 for EBV) in conjunctival SCC tissues, 3. Explore the location of EBV transcripts across different cell types (B-cells, T-cells and epithelial cells) in conjunctival SCC FFPE tissue samples, stratified by HPV co-infection and HIV status. The study design will be a case-control retrospective study, retrieving an estimated 200 consecutive cancer samples (cases) and 200 cancer free controls. Cases and controls will be matched by sex, age (+/- 10 years) and where possible HIV status with histologically confirmed diagnosis between 1 January 2015 and 31 January 2020 from a central tertiary histopathology laboratory in Harare Zimbabwe, where most provincial and private hospitals refer their samples to. Viral DNA and RNA will be detected in epithelial and/or B and T cells from FFPE samples by PCR-based assays and in situ-hybridization. Our study will build on our strong preliminary data and is anticipated to provide evidence for the involvement of a unique combination of infectious agents (EBV/HPV) in conjunctival SCC. Our findings will offer new opportunities for prevention and screening interventions aimed at populations in Africa in which this malignancy is more common.