Project 3-Cameron

项目总结/摘要 致癌性人乳头瘤病毒（高危型HPV，或hrHPV）是导致更多 90%以上的宫颈癌和肛管癌（肛门生殖器癌）。感染者 人免疫缺陷病毒（HIV）感染（PLWH）特别易受hrHPV介导 肛门生殖器癌感染高危型HPV的人患癌症的风险增加，如果病毒基因组 整合到宿主基因组中。hrHPV整合促进肿瘤发生的机制 已经得到了很好的描述;然而，促进hrHPV整合的因素在很大程度上尚不清楚。的 建议研究的长期目标是确定这些因素，然后设计有效的 采取干预措施，预防艾滋病毒携带者患上肛门生殖器癌。研究表明， 患有肛门生殖器高危型HPV感染并同时感染另一种肿瘤病毒--EB病毒的患者 (EBV)，患癌前发育不良的可能性是患有肛门生殖器发育不良的人的3-6倍。 hrHPV在不存在EBV的情况下。该提案调查了EBV促进整合的假设 通过造成DNA损伤将hrHPV基因组导入宿主基因组。一般实验方法 将利用从hrHPV感染的女性和男性收集的宫颈和肛门组织， 了解肛门生殖器组织中hrHPV和EBV之间的相互作用。的空间关系 在共感染组织中hrHPV和EBV之间的相互作用是HPV感染机制的本质的关键决定因素。 病毒之间的相互作用;然而，迄今为止的研究未能确定细胞隔室 在肛门生殖器组织中被EB病毒占据。因此，本提案的第一个具体目标是确定 hrHPV和EBV通过共同感染同一细胞直接相互作用，或者如果它们通过感染 使用尖端原位杂交和免疫组化技术， 化学接近。第二个具体目标是开始阐明hrHPV-EBV共表达的作用。 感染宿主和病毒基因表达网络。hrHPV-EBV共感染中的转录表达 将使用高深度转录组学将肛门生殖器组织与hrHPV感染的组织进行比较 方法.宿主转录本的差异表达将揭示hrHPV干扰的细胞途径- EBV合并感染，包括合并感染对DNA损伤/修复途径的影响。病毒基因 a）EBV是否表达与DNA相关的潜伏转录物 损伤和肿瘤发生; B）hrHPV是否更可能在EBV存在下整合;和c）如果 EBV诱导hrHPV癌基因的表达。这些目标将共同为工作模型提供信息 描述了肛门生殖器癌背景下hrHPV和EBV之间的机制相互作用 这将在随后的R 01提案中进一步探讨。这些研究将阐明 新的干预策略的机会，以防止肛门生殖器癌的PLWH。

PROJECT SUMMARY / ABSTRACT Oncogenic human papillomavirus (high risk HPV, or hrHPV) is the etiologic agent responsible for more than 90% of cancers of the uterine cervix and the anal canal (anogenital cancers). People living with human immunodeficiency virus (HIV) infection (PLWH) are particularly susceptible to hrHPV mediated anogenital cancers. People infected with high-risk HPV are at increased risk for cancer if the viral genome integrates into the host genome. The mechanisms by which hrHPV integration promotes tumorigenesis have been well described; however, the factors that promote hrHPV integration are largely unknown. The long-term objective of the proposed research is to identify these factors and then design effective interventions to prevent PLWH from developing anogenital cancers. Research has shown that PLWH who have anogenital high-risk HPV infection concurrently with another tumor virus, Epstein-Barr virus (EBV), are 3-6 times more likely to have pre-cancerous dysplasia than people who have anogenital hrHPV in the absence of EBV. This proposal investigates the hypothesis that EBV promotes integration of hrHPV genomes into the host genome by causing DNA damage. The general experimental approach will utilize cervical and anal tissues collected from hrHPV infected women and men to gain a better understanding of the interactions between hrHPV and EBV in anogenital tissue. The spatial relationship between hrHPV and EBV in co-infected tissue is a key determinant of the nature of the mechanistic interactions between the viruses; however, studies to date have failed to identify the cell compartment occupied by EBV in anogenital tissue. Therefore, the first Specific Aim of this proposal is to determine if hrHPV and EBV interact directly by co-infecting the same cell, or if they interact indirectly by infecting proximal but separate cell compartments, using cutting-edge in situ hybridization and immunohisto- chemistry approaches. The second Specific Aim is to begin to elucidate the effects of hrHPV-EBV co- infection on host and virus gene expression networks. Transcript expression in hrHPV-EBV co-infected anogenital tissue will be compared to that of hrHPV infected tissue using high-depth transcriptomics methods. Differential expression of host transcripts will reveal cellular pathways perturbed by hrHPV- EBV co-infection, including the effects of co-infection on DNA damage/repair pathways. Viral gene expression will be observed to determine a) if EBV is expressing latency transcripts associated with DNA damage and oncogenesis; b) if hrHPV is more likely to be integrated in the presence of EBV; and c) if EBV induces the expression of hrHPV oncogenes. Together these aims will inform the working model describing the mechanistic interactions between hrHPV and EBV in the context of anogenital cancer development, which will be further explored in a subsequent R01 proposal. These studies will illuminate opportunities for novel intervention strategies to prevent anogenital cancers in PLWH.