(PQ5) Role of HIV-associated cellular miRNAs in HPV 16-induced pathogenesis

 描述（由申请人提供）：该项目的重点是了解为什么HIV阳性与HIV阴性个体更有可能进展为人乳头瘤病毒16（HPV 16）相关的肛门高级鳞状上皮内病变（HSIL）和肛门癌。虽然HPV相关的肛门HSIL和来自HIV阳性和HPV阴性个体的癌症在组织病理学上相似，但是在感染HPV之前或之后，上皮暴露于HIV可能导致上皮中的细胞变化，这更可能导致细胞凋亡。 转化，从而增加肛门癌的发病率在这一人群中。 一些研究表明，外源性HIV蛋白（例如达特和nef）干扰已知在HPV 16相关癌症中失调的细胞蛋白和途径。达特和nef干扰细胞蛋白及其相应途径的一种方式是通过调节microRNA（miRNA）表达。人类基因组编码超过3，707种miRNA，这些miRNA经常靶向与癌症发展或预防相关的许多基因。已经在几种类型的肿瘤和这些肿瘤的不同阶段检测到差异表达的miRNA。我们假设HIV蛋白调节肛门上皮细胞的miRNA谱，导致调节宿主癌基因和肿瘤抑制基因的细胞通路失调。与HPV 16共感染进一步增强了细胞miRNA及其相应途径的这种失调。 与肛门肿瘤诊所，研究和教育（ANCRE）和加州大学旧金山分校的桑德勒功能基因组学设施合作，我们将在HIV阳性和HIV阴性个体的存档肛门活检中进行miRNA分析。我们将使用HTG EdgeSeq miRNA全转录组测定法比较不同病理级别组织中的细胞miRNA谱，包括正常组织，HSIL和癌症，HTG EdgeSeq miRNA全转录组测定法是一种测量2，275种人类miRNA表达的新技术。这将使我们能够描绘出HPV 16阳性肛门肿瘤的差异和相似性，这些肿瘤来自HIV阳性与HIV阴性个体。此外，我们将使用肛门上皮体外模型：1）鉴定导致细胞miRNA失调的HIV蛋白，2）在HIV/HPV 16单独感染和合并感染的情况下检测一组感兴趣的miRNA的表达，3）证明单个miRNA对细胞通路和蛋白质的影响，这些通路和蛋白质参与HPV 16相关的肛门癌进展。 这将是第一项在HIV感染背景下建立肛门癌特异性miRNAome的研究。HIV相关细胞miRNAs的鉴定， HPV 16相关的肛门癌进展可能导致新的诊断/预后标志物的开发，以及治疗肛门癌的疗法，特别是在HIV阳性个体中。

 DESCRIPTION (provided by applicant): This project is focused on understanding of why HIV-positive versus HIV-negative individuals are more likely to progress to human papillomavirus 16 (HPV 16)-associated anal high-grade squamous intraepithelial lesions (HSIL) and anal cancer. Although HPV-associated anal HSIL and cancers from HIV-positive and HPV- negative individuals are histopathologically similar, epithelial exposure to HIV, before or after infection with HPV, may lead to cellular changes in the epithelium that are more likely to result in cellular transformation, thus increasing the incidence of anal cancer in this population. Several studies have suggested that exogenous HIV proteins (e.g. tat and nef) interfere with cellular proteins and pathways that are known to be dys-regulated in HPV 16-associated cancers. One way that tat and nef interfere with cellular proteins and their corresponding pathways is through modulation of microRNA (miRNA) expression. The human genome encodes over 3,707 miRNAs, which frequently target many genes related to cancer development or prevention. Differentially-expressed miRNAs have been detected in several types of tumors and at different stages of those tumors. We hypothesize that HIV proteins modulate the miRNA profile of anal epithelial cells leading to dys-regulation of cellular pathways that regulate host oncogenes and tumor suppressor genes. Co-infection with HPV 16 further enhances this dys-regulation of cellular miRNAs and their corresponding pathways. Partnered with the Anal Neoplasia Clinic, Research and Education (ANCRE) and the Sandler Functional Genomics Facility at UCSF, we will perform miRNA profiling in archived anal biopsies from HIV- positive and HIV-negative individuals. We will compare the cellular miRNA profiles in tissues of different grades of pathology, including normal tissues, HSIL and cancer using the HTG EdgeSeq miRNA whole transcriptome assay, a novel technique that measures the expression of 2,275 human miRNAs. This will allow us to delineate the differences and similarities in HPV 16-positive anal tumors that arise from HIV-positive versus HIV-negative individuals. In addition we will use an anal epithelial in-vitro model to: 1) Identify HIV proteins that lead to dys-regulation of cellular miRNAs, 2) Validate the expression of a subset of miRNAs of interest in the setting of HIV/HPV 16 infection alone and during co-infection, and 3) Demonstrate the effects of individual miRNAs on cellular pathways and proteins that are involved in HPV 16-associated anal cancer progression. This will be the first study to establish a miRNAome specific to anal cancer in the setting of HIV infection. The identification of HIV-associated cellular miRNAs that are involved in HPV 16-associated anal cancer progression could lead to development of new diagnostic/prognostic markers, as well as therapies to treat anal cancers, particularly in HIV-positive individuals.