Regulation of ZEB1 by Porphyromonas gingivalis in gingival epithelial cells

牙龈上皮细胞中牙龈卟啉单胞菌对ZEB1的调节

基本信息

批准号：
10228577
负责人：
Zackary R Fitzsimonds
金额：
$ 4.34万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10228577
关键词：
Amanitins Anaerobic Bacteria Apical Apoptosis Binding Biopsy Blood Circulation Cancer Patient Cell Cycle Cell Cycle Progression Cell Nucleus Cells Co-Immunoprecipitations Complex Data Development Diagnosis E-Cadherin E1A-associated p300 protein Embryonic Development Environment Epidemiology Epithelial Epithelial Cells Esophageal Squamous Cell Carcinoma Family Foundations Future Gene Expression Genes Genetic Transcription Germ Cells Gingiva Homeobox Human Immunofluorescence Immunologic Immunohistochemistry Immunoprecipitation Intercept Invaded Knowledge Laboratories Lead Literature Luciferases MADH2 gene MADH3 gene MADH4 gene MADH6 gene MADH7 gene MMP9 gene Malignant Neoplasms Measures Mediating Mesenchymal MicroRNAs Migration Assay Molecular N-Cadherin Neoplasm Metastasis Nuclear Oncogenic Oral Oral cavity Pathway interactions Patients Periodontal Diseases Periodontitis Phenotype Phosphorylation Plasmids Play Porifera Porphyromonas gingivalis Primary carcinoma of the liver cells Process Prognosis Property Quantitative Reverse Transcriptase PCR RNA RNA Synthesis Inhibitors Ramp Regulation Role Sampling Signal Pathway Signal Transduction Speed Survival Rate TGF Beta Signaling Pathway Testing Tight Junctions Time Tongue Neoplasms Transcription Coactivator Transforming Growth Factor beta Receptors Translating Untranslated RNA Up-Regulation Western Blotting Zinc Fingers arm cancer cell cancer diagnosis differential expression dysbiosis epithelial to mesenchymal transition expression vector genetic corepressor host-microbe interactions human tissue knock-down mRNA Expression mRNA Stability malignant mouth neoplasm mouth squamous cell carcinoma new therapeutic target novel novel diagnostics novel therapeutics overexpression prevent promoter recruit response scaffold synergism transcription factor tumor

项目摘要

Project Summary Oral cancer is the 11th most frequently diagnosed cancer in the world, and due to its late diagnosis, it has an approximately 50% survival rate after 5 years. Metastasis is meditated through Epithelial to Mesenchymal Transition (EMT), which is the process by which epithelial cells lose their tight junctions and apical/basal polarity and shift to a more invasive mesenchymal phenotype giving the cancer cells systemic access through invasion of the bloodstream. EMT is mediated through several transcriptional factors, however we will focus on ZEB1. Porphyromonas gingivalis is an oral obligate anaerobe that when present shifts the host/microbe relationship from synergy to a more dysbiotic environment, which contributes to periodontal disease. P. gingivalis has been shown to invade gingival epithelial cells, and once inside the cells can inhibit apoptosis, ramp up the cell cycle, and more recently by our lab has been shown to upregulate ZEB1. Although we are beginning to understand the potential oncogenic role of P. gingivalis, we do not currently know how ZEB1 is regulated by P. gingivalis in gingival epithelial cells. Our preliminary data suggests that this regulation is occurring through SMAD signaling regulation and through the long non-coding RNA ZEB1-AS1. SMAD signaling is a branch of the TGF-b signaling pathway, which during embryonic development is responsible for producing mesenchymal cells from the initial epithelium. SMAD signaling occurs by phosphorylation of SMAD2/3 by the TGF-b receptor, followed by the formation of a transcriptional regulatory trimer by SMAD2/3 and SMAD4, and finally nuclear localization and upregulation of ZEB1. Additionally, ZEB1 can bind to SMAD2/3 and SMAD4 to form a ZEB1/SMAD trimer to regulate ZEB1 transcription. We have found that P. gingivalis upregulates the regulatory SMAD3, which suggests P. gingivalis is able to intercept this pathway to upregulate ZEB1. We have also found that P. gingivalis upregulates ZEB1-AS1, which has been shown in hepatocarcinoma and esophageal squamous cell carcinoma to directly upregulate ZEB1, and our preliminary data supports this as knocking down ZEB1-AS1 leads to abrogation of ZEB1 regulation by P. gingivalis in gingival epithelial cells. We will investigate mechanisms of regulation by ZEB1-AS1 including direct promotion of gene expression in the nucleus through binding to P300, and/or control of ZEB1 mRNA stability. EMT regulation is a complex process that involves integration of several host signaling pathways, so we also suspect that ZEB1-AS1 will be integrated with SMAD signaling by potentially acting as a scaffold for SMAD3. Developing a better understanding of P. gingivalis mediated EMT will lay the foundation for future studies to target these regulatory mechanisms for diagnosis and treatment, and ultimately reduce the 50% chance of survival current patients have 5 years after diagnosis.

项目概要口腔癌是世界上第 11 位最常诊断的癌症，由于诊断较晚，它的发病率很高。 5年后生存率约为50%。转移是通过上皮细胞到间质细胞进行的转变 (EMT)，这是上皮细胞失去紧密连接和顶端/基底极性的过程并转变为更具侵袭性的间充质表型，使癌细胞能够通过侵袭进入全身的血液。 EMT 通过多种转录因子介导，但我们将重点关注 ZEB1。牙龈卟啉单胞菌是一种口腔专性厌氧菌，当其存在时会改变宿主/微生物的关系从协同作用到更加失调的环境，这会导致牙周病。牙龈卟啉单胞菌显示可侵入牙龈上皮细胞，一旦进入细胞即可抑制细胞凋亡，加速细胞周期，最近我们的实验室已证明可以上调 ZEB1。尽管我们刚刚开始了解牙龈卟啉单胞菌的潜在致癌作用，我们目前不知道牙龈卟啉单胞菌如何调节ZEB1 牙龈上皮细胞。我们的初步数据表明这种调节是通过 SMAD 信号传导发生的调节并通过长非编码RNA ZEB1-AS1。 SMAD 信号传导是 TGF-b 信号传导的一个分支途径，在胚胎发育过程中负责从最初的细胞产生间充质细胞上皮。 SMAD 信号传导通过 TGF-b 受体磷酸化 SMAD2/3 来发生，然后 SMAD2/3 和 SMAD4 形成转录调控三聚体，最后核定位和 ZEB1 的上调。此外，ZEB1 可以与 SMAD2/3 和 SMAD4 结合形成 ZEB1/SMAD 三聚体，调节ZEB1转录。我们发现牙龈卟啉单胞菌上调调节性 SMAD3，这表明 P. gingivalis 能够拦截该途径来上调 ZEB1。我们还发现牙龈卟啉单胞菌上调 ZEB1-AS1，已在肝癌和食管鳞状细胞癌中得到证实直接上调ZEB1，我们的初步数据支持这一点，因为敲低ZEB1-AS1会导致牙龈上皮细胞中牙龈卟啉单胞菌对ZEB1调节的废除。我们将研究机制 ZEB1-AS1 的调节包括通过与 P300 结合直接促进细胞核中的基因表达，和/或ZEB1 mRNA稳定性的控制。 EMT 监管是一个复杂的过程，涉及多个方面的整合宿主信号通路，因此我们也怀疑 ZEB1-AS1 可能通过潜在的方式与 SMAD 信号通路整合作为 SMAD3 的支架。更好地了解牙龈卟啉单胞菌介导的 EMT 将为为未来研究针对这些调节机制进行诊断和治疗奠定基础，并最终当前患者诊断后 5 年的生存机会减少 50%。