Investigating the mechanism by which PTPN14 degradation by HPV E7 represses epithelial differentiation

研究 HPV E7 降解 PTPN14 抑制上皮分化的机制

• 批准号: 10326841
• 负责人: Joshua Hatterschide
• 金额: $ 4.21万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-06-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10326841
• 关键词: Amino Acid Motifs; Anogenital cancer; Award; Basal Cell; Binding; Biochemistry; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cell physiology; Collaborations; Complex; Confocal Microscopy; Core Facility; Data; Elements; Epithelial; Epithelial Cells; Extracellular Matrix; Foundations; Genetic Transcription; Genotype; Goals; HPV E7; Human Activities; Human Papillomavirus; Human papilloma virus 31; Human papilloma virus infection; Hydrogels; Immunoprecipitation; Knock-out; Life Cycle Stages; Maintenance; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Mass Spectrum Analysis; Mediating; Modeling; Oncogenes; Oncogenic; Oncoproteins; Oral; Phase; Physiological; Play; Positioning Attribute; Postdoctoral Fellow; Protein Tyrosine Phosphatase; Proteins; Proteomics; Publishing; Regulation; Repression; Research; Research Personnel; Role; Scaffolding Protein; Signal Transduction; Stratified Epithelium; Stratified Squamous Epithelium; Stratum Basale; Techniques; Testing; Training; Transcription Coactivator; Tumor Suppressor Proteins; Undifferentiated; Viral; Viral Physiology; Viral reservoir; Virus Latency; carcinogenicity; chronic infection; experience; genetic regulatory protein; keratinocyte; latent infection; malignant oropharynx neoplasm; oral cavity epithelium; overexpression; paralogous gene; prevent; programs; skills; tissue culture; tumorigenesis; virus host interaction

PROJECT SUMMARY/ABSTRACT: Persistent infection with oncogenic genotypes of human papillomavirus (HPV) can cause oropharyngeal and anogenital cancers. The HPV E7 oncoprotein is both a central driver of HPV-mediated oncogenesis and an important accessory protein for sustaining HPV infection. This proposal focuses on a highly conserved activity of HPV E7: degradation of the host protein tyrosine phosphatase, PTPN14. It was recently discovered that PTPN14 degradation by HPV E7 repress epithelial differentiation. PTPN14 was previously not characterized as a regulator of differentiation, and repression of differentiation through PTPN14 is significant because HPV+ cancers are often poorly differentiated. In an HPV infected stratified epithelium, HPV establishes the stable maintenance phase of its life cycle undifferentiated basal keratinocytes, and HPV exits this phase of its life cycle and commits to productive replication when an infected cell differentiates. Therefore, repression of differentiation through PTPN14 degradation by HPV E7 is also significant because it could play a role in regulating the transition between these phases of the HPV life cycle. However, the mechanism by which PTPN14 degradation by HPV E7 represses differentiation is not defined, and the impact of repressing differentiation on the HPV life cycle has yet to be identified. PTPN14 is a tumor suppressor that can inhibit the transcriptional regulators Yes-associated protein (YAP) and TAZ. PTPN14 can also bind to the scaffolding protein, KIBRA, to inhibit YAP/TAZ. YAP/TAZ are oncogenes that promote epithelial basal cell identity and repress differentiation. Therefore, PTPN14 degradation by HPV E7 may repress differentiation by disrupting PTPN14- and KIBRA-containing signaling complexes and activating YAP/TAZ. Preliminary data gathered for this proposal implicate YAP/TAZ and KIBRA in mechanisms surrounding PTPN14 in keratinocytes. The objectives of this proposal are to determine the mechanism by which PTPN14 degradation by E7 represses differentiation and to determine how this mechanism impacts the HPV life cycle. The specific aims of this proposal are 1) to determine if PTPN14 degradation by HPV E7 activates YAP/TAZ and regulates the HPV life cycle, and 2) to determine if PTPN14 degradation by HPV E7 disrupts KIBRA- mediated signaling. Aim 1 will test if PTPN14 degradation by HPV E7 activates YAP/TAZ and promotes the transcription of YAP/TAZ transcriptional targets in oral epithelial keratinocytes. Furthermore, it will test how constitutively active YAP/TAZ impact the HPV life cycle. Aim 2 will test if PTPN14 requires KIBRA to induce differentiation in oral keratinocytes, and test whether PTPN14 degradation by HPV E7 disrupts KIBRA-containing signaling complexes. Defining this mechanism will augment our understanding of the HPV E7 oncoprotein, which has implications for both its carcinogenic activities and its role in HPV infection. Completing the proposed aims will provide training in several key skills, preparing me to pursue post-doctoral research following this award.

项目摘要/摘要： 人乳头瘤病毒 (HPV) 致癌基因型的持续感染可导致口咽部和 肛门生殖器癌症。 HPV E7 癌蛋白既是 HPV 介导的肿瘤发生的核心驱动因素，也是 维持 HPV 感染的重要辅助蛋白。该提案重点关注高度保守的活动 HPV E7：宿主蛋白酪氨酸磷酸酶 PTPN14 的降解。最近发现 HPV E7 导致的 PTPN14 降解抑制上皮分化。 PTPN14 之前并未被定性为 分化的调节因子，通过 PTPN14 抑制分化非常重要，因为 HPV+ 癌症通常分化较差。在 HPV 感染的复层上皮中，HPV 建立了稳定的 其生命周期的维持阶段未分化的基底角质形成细胞，HPV 退出其生命周期的此阶段 并在受感染的细胞分化时进行高效复制。因此，抑制分化 HPV E7 降解 PTPN14 也很重要，因为它可以在调节转变中发挥作用 HPV 生命周期的这些阶段之间。然而，HPV 降解 PTPN14 的机制 E7抑制分化尚未明确，抑制分化对HPV生命周期的影响已被证实 尚未确定。 PTPN14是一种肿瘤抑制因子，可以抑制转录调节因子Yes相关蛋白（YAP）和 塔兹。 PTPN14 还可以与支架蛋白 KIBRA 结合，抑制 YAP/TAZ。 YAP/TAZ 是致癌基因 促进上皮基底细胞身份并抑制分化。因此，PTPN14被HPV E7降解可能 通过破坏含有 PTPN14 和 KIBRA 的信号复合物并激活 雅普/塔兹。为本提案收集的初步数据表明 YAP/TAZ 和 KIBRA 参与了相关机制 角质形成细胞中的 PTPN14。该提案的目标是确定 PTPN14 的机制 E7 的降解抑制分化并确定该机制如何影响 HPV 生命周期。 该提案的具体目标是 1) 确定 HPV E7 降解 PTPN14 是否激活 YAP/TAZ 并调节 HPV 生命周期，2) 确定 HPV E7 降解 PTPN14 是否会破坏 KIBRA- 介导的信号传导。目标 1 将测试 HPV E7 降解 PTPN14 是否激活 YAP/TAZ 并促进 口腔上皮角质形成细胞中 YAP/TAZ 转录靶标的转录。此外，它将测试如何 持续活跃的 YAP/TAZ 影响 HPV 生命周期。目标 2 将测试 PTPN14 是否需要 KIBRA 来诱导 口腔角质形成细胞的分化，并测试 HPV E7 降解 PTPN14 是否会破坏含有 KIBRA 的 信号复合物。定义这一机制将增强我们对 HPV E7 癌蛋白的理解，该蛋白 其致癌活性及其在 HPV 感染中的作用都有影响。完成拟议目标 将提供多项关键技能的培训，为我在获奖后继续从事博士后研究做好准备。