Viral-host interactions that influence the life cycle of DNA tumor viruses

影响DNA肿瘤病毒生命周期的病毒-宿主相互作用

• 批准号: 10696205
• 负责人: CARY A MOODY
• 金额: $ 34.72万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10696205
• 关键词: Affect; Architecture; Cells; Cellular biology; Chromatin; Chromosomes; DNA; DNA Damage; DNA Tumor Viruses; DNA biosynthesis; Data; Development; Environment; Epithelial Cells; Epithelium; Exhibits; Future; Gene Expression; Genetic Transcription; Genomic Instability; Goals; HPV-High Risk; Herpesviridae; High-Throughput Nucleotide Sequencing; Human; Human Herpesvirus 4; Human Herpesvirus 8; Human Papilloma Virus Vaccine; Human Papilloma Virus-Related Malignant Neoplasm; Human Papillomavirus; Human papilloma virus 31; Human papilloma virus infection; Hybrids; Infection; Knowledge; Lesion; Life Cycle Stages; Link; Location; Lytic Phase; Lytic Virus; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Maps; Modification; Oncogenic Viruses; Oncoproteins; Oral; Oral cavity; Oropharyngeal; Oropharyngeal Squamous Cell Carcinoma; Pathogenesis; Pathway interactions; Production; Productivity; Proliferating; RNA; Regulation; Role; Saliva; Source; Squamous cell carcinoma; Stratified Epithelium; Subcellular Anatomy; Testing; Undifferentiated; Untranslated RNA; Viral; Viral Genome; Viral Pathogenesis; Virion; Virus; Virus Replication; Work; carcinogenesis; cell type; differential expression; high risk; insight; keratinocyte; low and middle-income countries; lytic replication; malignant mouth neoplasm; new therapeutic target; novel strategies; oral cavity epithelium; overexpression; premalignant; programs; replication stress; therapeutic development; transcriptome; transmission process; tumorigenesis; viral DNA; virus host interaction; virus related cancer

Project 2 Project Summary Abstract Oncogenic viruses cause 15-20% of human cancers. High-risk HPVs are associated with 5% of cancers, including cervical cancer (CC) as well as an increasing number of oropharyngeal squamous cell carcinomas (OSCC). RNA molecules, such as long non-coding RNAs (lncRNAs) and co-transcriptional R-loops (RNA:DNA hybrids) are associated with genomic instability and other hallmarks of cancer, but are understudied in the context of HPV infection. Our preliminary data indicate high-risk HPV31 positive cells have increased levels of R-loops compared to uninfected cells. Furthermore, we have identified several lncRNAs that are altered by HPV31 that influence epithelial differentiation, proliferation and survival. Whether these RNA molecules serve a pro-viral role and/or contribute to HPV pathogenesis is unclear. The DNA tumor viruses EBV and KSHV are also found in the oral cavity and cause oral cancers. Similarly to HPV, EBV and KSHV also use differentiation of the oral epithelium as a mechanism to activate lytic replication and virus production. Oral epithelial cells are the likely source of infectious virus in the saliva and a critical component of viral pathogenesis. However, the mechanisms that regulate the productive replication of HPV, as well as the latent/lytic phases of EBV and KSHV in the oral epithelium are not well characterized. These viruses likely employ similar strategies to uncouple proliferation from differentiation to promote both viral persistence and viral spread. KSHV and EBV alter the host lncRNA profile in other cell types, and R-loop forming sequences have been identified in KSHV and EBV viral genomes. Additionally, we have found that KSHV induces R-loop formation and DNA damage in oral epithelial cells. We hypothesize that HPV, EBV and KSHV reprogram the epithelial cell environment through R-loop formation/distribution and lncRNA expression in order to support the viral life cycle. Specific Aims to test this hypothesis are: (1) Determine the role of R-loops in HPV and KSHV pathogenesis by mapping R-loop distribution on cellular and viral DNA and determining if R-loops contribute to DNA damage in infected cells; (2) Determine how cellular lncRNAs contribute to the life cycle of HPV as well as KSHV and EBV in epithelial cells through a biased approach of overexpression/depletion studies of specific lncRNAs, and an unbiased approach through high throughput sequencing to examine global alterations in lncRNA expression. Understanding how R-loops and lncRNAs contribute to replication of these DNA tumor viruses may uncover a role for RNA molecules in facilitating viral persistence as well as identify cellular pathways that may be exploited for the treatment of viral- and potentially non-viral-associated cancers.

计划2 项目摘要 致癌病毒导致15-20%的人类癌症。高危HPV与5%的癌症有关， 包括宫颈癌（CC）以及越来越多的口咽鳞状细胞癌 （口腔鳞状细胞癌）。RNA分子，例如长非编码RNA（lncRNA）和共转录R环（RNA：DNA 杂交）与基因组不稳定性和其他癌症标志有关，但在 HPV感染的背景。我们的初步数据表明，高危HPV 31阳性细胞具有增加的水平， 与未感染细胞相比的R环。此外，我们已经确定了几种lncRNA， HPV 31影响上皮分化、增殖和存活。这些RNA分子是否作为 促病毒作用和/或促成HPV发病机制尚不清楚。DNA肿瘤病毒EBV和KSHV也是 在口腔中发现并导致口腔癌。与HPV类似，EBV和KSHV也使用细胞分化。 口腔上皮作为激活裂解复制和病毒产生的机制。口腔上皮细胞很可能是 唾液中的传染性病毒来源和病毒发病机制的关键组成部分。然而，机制 调节HPV的生产性复制，以及口腔中EBV和KSHV的潜伏/裂解期。 上皮没有被很好地表征。这些病毒可能采用类似的策略来解偶联增殖 从分化到促进病毒持久性和病毒传播。KSHV和EBV改变宿主lncRNA 在其他细胞类型中，R-loop形成序列已在KSHV和EBV病毒基因组中鉴定。 此外，我们发现KSHV诱导口腔上皮细胞中的R环形成和DNA损伤。我们 假设HPV、EBV和KSHV通过R-loop重编程上皮细胞环境 病毒的形成/分布和lncRNA的表达，以支持病毒的生命周期。具体目的是测试这个 假设：（1）通过绘制R-loop分布图确定R-loop在HPV和KSHV发病机制中的作用 细胞和病毒DNA上，并确定R环是否有助于感染细胞中的DNA损伤;（2）确定 细胞lncRNA如何通过一种新的途径促进HPV以及KSHV和EBV在上皮细胞中的生命周期， 特异性lncRNA的过度表达/缺失研究的偏倚方法，以及通过 高通量测序以检查lncRNA表达的总体改变。了解R环如何 和lncRNA有助于这些DNA的复制肿瘤病毒可能揭示了RNA分子在 促进病毒持久性以及鉴定可用于治疗病毒性结肠炎的细胞途径。 和潜在的非病毒相关癌症。