Forkhead factor regulation in oral viral infection

口腔病毒感染中叉头因子的调节

基本信息

批准号：
10425257
负责人：
Natalie Atyeo
金额：
$ 4.34万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2023-05-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10425257
关键词：
Address Affect Binding Binding Sites Biological Biological Assay Cell Differentiation process Cell Proliferation Cell physiology Cells Chromatin Chronic Clinical Clinical Research Complex Coupled DNA Binding Data Dentistry Deposition Differentiation and Growth EZH2 gene Enhancers Environment Enzymes Epigenetic Process Epithelial Cells Exhibits FOXQ1 gene Family Florida Gene Expression Genes Genetic Transcription Genomic approach Gingiva Herpesviridae Herpesviridae Infections Herpesvirus 1 Hour Human Human Herpesvirus 8 Human body Immune response Individual Infection Integration Host Factors KDM1A gene Laboratories Lead Life Luciferases Lytic Lytic Phase Malignant Neoplasms Mediating Mediator of activation protein Mentorship Methylation Notch Signaling Pathway Nucleic Acid Regulatory Sequences Oncogenic Oncogenic Viruses Oral Oral cavity Palate Kaposi&apos s Sarcoma Pathogenesis Pathway interactions Primary Infection Production Proteins Regulation Repression Research Research Training Role Route Saliva Scientist Site Small Interfering RNA Testing Training Training Programs Transcription Coactivator Universities Viral Viral Genes Viral Genome Viral Pathogenesis Viral Proteins Virion Virus Virus Diseases Virus Replication Virus Shedding base career cell growth cell type chromatin immunoprecipitation chronic infection college gene repression genome-wide genome-wide analysis latent infection member mutant notch protein novel oral biology oral cavity epithelium oral infection pathogen promoter response skills small molecule inhibitor transcription factor transcriptome transcriptome sequencing transmission process virology

项目摘要

The oral cavity is the initial site of infection for many pathogens, including viruses. Kaposi's sarcoma-associated herpesvirus (KSHV) is an oncogenic virus that is shed in saliva, which is the primary route for spread between individuals. While in most cell types KSHV establishes a lifelong latent infection, in primary human gingival epithelial cells (HGEP) KSHV can establish a lytic infection, with sustained viral gene expression driven by the essential lytic viral factor replication and transcription activator (RTA). Importantly, lytic KSHV infection of oral epithelial cells leads to the production and release of new infectious virions, a key early step in KSHV infection that can lead to viral spread and chronic infection of other cell types in the human body. Despite the clinical and biological importance of oral infection as an initial step in KSHV infection, little is known about the host-pathogen interactions and their control mechanisms during KSHV infection of oral epithelial cells. Through genome-wide transcriptome analysis, one member of the forkhead box family of transcription factors (FOX TFs) was identified as rapidly and robustly induced upon de novo KSHV infection in HGEP cells. The FOX factors comprise a family TFs responsible for many cellular processes including cell growth and differentiation. However, little is known about the roles of FOX TFs in the context of viral infection. Preliminary data revealed that a large number of predicted target genes of the FOX TF are de-regulated during KSHV infection. Many of these targets encode key regulators of critical cellular functions. While the identified FOX TF is highly induced upon KSHV infection of oral cells, it is expressed at very low levels in uninfected oral cells, suggesting a rapid transcriptional switch driven by viral infection. The purpose of this project is to dissect the mechanism of FOX TF silencing in uninfected oral epithelial cells, and to determine how lytic viral infection disrupts this silencing, thereby promoting rapid alterations of normal cellular functions in infected oral epithelial cells. To examine repressive mechanisms in the oral cavity, Aim 1 focuses on identification of epigenetic enzymes that restrict FOX TF expression. It is hypothesized that viral factors disrupt the silencing of these host epigenetic enzymes during lytic infections. Aim 2 will identify how specific KSHV proteins induce the expression of FOX TF. A combination of genome-wide studies coupled with KSHV mutants will be used to analyze how KSHV infection affects FOX TF regulatory regions to result in FOX TF induction. In summary, this study will identify a novel viral mechanism in which the virus exploits a central host transcription factor to deregulate key cellular processes during lytic infection. The completion of this project will provide in-depth training in epigenetics, virology and genomics approaches. These skills will be enhanced in the outstanding research environment provided by the Comprehensive Training Program in Oral Biology at the University of Florida College of Dentistry, which incorporates tailored mentorship and excellent clinical and research training to prepare the applicant for a career as a clinician-scientist.

口腔是许多病原体（包括病毒）的初始感染部位。卡波西与肉瘤相关疱疹病毒（KSHV）是一种在唾液中脱落的致癌病毒，这是在之间传播的主要途径个人。尽管在大多数细胞类型中，KSHV建立了终生的潜在感染，但在原发性牙龈中上皮细胞（HGEP）KSHV可以建立裂解感染，并具有持续的病毒基因表达。必需的裂解病毒因子复制和转录激活剂（RTA）。重要的是，口服裂解KSHV感染上皮细胞导致新的感染性病毒体的产生和释放，这是KSHV感染的关键早期一步这可能导致人体其他细胞类型的病毒扩散和慢性感染。尽管有临床和口腔感染作为KSHV感染的第一步的生物学重要性，对宿主病原体知之甚少在口腔上皮细胞KSHV感染期间的相互作用及其控制机制。通过全基因组转录组分析，确定了转录因子家族（FOX TF）的一个成员在HGEP细胞中从头kSHV感染而迅速且稳健地诱导。狐狸因素包括一个家庭 TF负责许多细胞过程，包括细胞生长和分化。但是，鲜为人知关于FOX TF在病毒感染中的作用。初步数据显示了大量在KSHV感染期间，FOX TF的预测靶基因被解调。这些目标中有许多编码关键细胞功能的关键调节剂。虽然在KSHV感染后，已确定的FOX TF被高度诱导口服细胞，在未感染的口腔细胞中以非常低的水平表达，这表明转录开关快速由病毒感染驱动。该项目的目的是剖析未感染的FOX TF沉默的机制口服上皮细胞，并确定裂解病毒感染如何破坏这种沉默，从而促进快速受感染的口腔上皮细胞中正常细胞功能的改变。检查抑制性机制口腔，AIM 1专注于识别限制FOX TF表达的表观遗传酶。这是假设病毒因子破坏了裂解感染期间这些宿主表观遗传酶的沉默。目的 2将确定特定的KSHV蛋白如何诱导FOX TF的表达。全基因组的结合结合KSHV突变体的研究将用于分析KSHV感染如何影响FOX TF调节导致FOX TF诱导的区域。总而言之，这项研究将确定一种新型的病毒机制，其中病毒利用中央宿主转录因子在裂解感染过程中放大管制细胞过程。这该项目的完成将提供对表观遗传学，病毒学和基因组学方法的深入培训。这些在综合培训提供的杰出研究环境中，技能将得到增强佛罗里达大学牙科学院口腔生物学课程，该学院纳入了量身定制的指导以及出色的临床和研究培训，为申请人做好准备的临床医生职业。