Role of Vimentin in Pathogenesis of CNS Inflammation

波形蛋白在中枢神经系统炎症发病机制中的作用

• 批准号: 8561170
• 负责人: SHENG-HE HUANG
• 金额: $ 24.3万
• 依托单位: CHILDREN'S HOSPITAL OF LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-15 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8561170
• 关键词: Bacterial Meningitis; Binding; Biological Markers; Blood; Blood - brain barrier anatomy; Brain; Cause of Death; Cell Adhesion; Cell Nucleus; Cells; Complex; Disease; Dissociation; Endothelial Cells; Escherichia coli; Gene Expression; Genes; Genetic; Goals; Head; Health; Human; In Vitro; Infection; Inflammation; Interphase Cell; Lead; Leukocytes; Low Birth Weight Infant; Mediating; Membrane Microdomains; Meningitis; Microbial Biofilms; Molecular Profiling; Monitor; Multi-Drug Resistance; Mus; NF-kappa B; Newborn Infant; Nuclear Translocation; PDZ protein; Pathogenesis; Pathway interactions; Penetration; Permeability; Phosphotransferases; Play; Proteins; Role; Signal Transduction; Signaling Molecule; Small Interfering RNA; Stem cells; Surface; System; Time; Tubulin; Ubiquitin; Up-Regulation; Vimentin; Virulence; Virulence Factors; base; bone; calmodulin-dependent protein kinase II; caveolin 1; central nervous system injury; early onset; genome-wide; indexing; inhibitor/antagonist; insight; microbial; migration; multicatalytic endopeptidase complex; neonatal sepsis; neutrophil; novel; pathogen; receptor; repaired; vimentin kinase

DESCRIPTION (provided by applicant): The E. coli IbeA receptor Vimentin (Vim), in conjunction with its co-receptor PSF, plays an important role in the pathogenesis of neonatal sepsis and meningitis (NSM), which remains a major cause of death in newborns, especially in low-birth weight infants. The ibeA locus is able to modulate expression of several virulence factors (e.g., fim, ibeB, ompA and biofilm-associated genes) and predominantly contribute to E. coli K1-caused early-onset human NSM by inducing both pathogen penetration and polymorphonuclear neutrophil (PMN) transmigration (PMNT) across the blood-brain barrier (BBB), which consists mainly of brain microvascular endothelial cells (BMEC). Vim has emerged as an organizer of a number of critical proteins involved in cell adhesion/migration and cell signaling. Vim- and PSF-mediated signaling is required for IbeA-induced NF-kB activation, pathogen penetration and PMNT across the BBB, which are the three hallmark features of bacterial meningitis. Lipid rafts (LRs) serve as a signaling platform for Vim, PSF, ¿7 nAChR (regulator of inflammation), and other signaling molecules (e.g., caveolin-1, ERK and CaM kinase II). Our recent studies have demonstrated for the first time that Vim and PSF cooperatively regulate IbeA+ E. coli K1-induced NF-kB signaling. Vim forms a complex with IkB, NF-kB and tubulins in the resting cells through its head domain (HD). Vim- mediated modulation of the ubiquitin proteasome system (UPS) is essential for NF-kB activation. IbeA is able to induce the IkB kinase (IKK) activation, dissociation of the Vim/IkB-NF-kB complex, and expression of Vim, ¿7 nAChR and proinflammatory factors (PIFs). Nuclear translocation of NF-kB is correlated with increased PSF in the nucleus. Circulating BMECs (cBMEC) and endothelial progenitor cells (EPC) in the blood could be used as cell-based biomarkers for indexing of the CNS injury and for monitoring genome-wide expression signatures (GES) during NSM. Most importantly, we have shown that IbeA+ E. coli-induced NF-kB activation, bacterial invasion, PMNT and BBB permeability are significantly reduced in Vim KO (-/-) mice, which are consistent with the in vitro findings. Based on these results, we hypothesize that Vim- and PSF-induced signaling is a new paradigm in which microbial factors (e.g., IbeA) induce cytoplasmic activation and nuclear translocation of NF-kB through the formation of the initial signaling complexes with other signaling molecules (e.g., caveolin-1, PI3K, ERK, CaM kinase II and Rac1) in lipid rafts. Subsequent activation of the IKK/NF-kB pathway and eventual upregulation of Vim, ¿7 nAChR and PIFs lead to CNS inflammation/BBB disorders, which can be detected by cBMEC/EPC-based approaches. Our hypothesis will be examined with the following specific aims: 1). Examine how Vim and PSF act in concert to modulate IbeA-induced NF-kB activation/translocation and how NF-kB subsequently up-regulates gene expression by defining Vim- and PSF-mediated signaling. 2). Determine how Vim and PSF contribute to IbeA-induced pathogen penetration and PMN transmigration across the BBB by SCP analysis of cBMEC and genetic blockage of Vim.

描述（由适用提供）：大肠杆菌IBEA受体波形蛋白（VIM）与其共受体PSF结合使用，在新生儿脓毒症和脑膜炎（NSM）的发病机理中起着重要作用，该作用仍然是新生儿的主要死亡原因，尤其是在低次体重婴儿中。 IBEA基因座能够调节多种病毒因子（例如FIM，IBEB，OMPA和生物膜相关基因），并主要促进大肠杆菌k1引起的早期爆发的人NSM，通过诱导病原体渗透和脑核中性粒细胞（PMN）（PMN）的脑（PMBR）（PMBR）（PMBR-BBR-BBR-BBR）（PMBR-BBR）（PMBR-BBR）（PMBR-BBR-BBR）的跨性别（PMBR）（PMBR），该（PMBR-BBR）跨越了bbbr，微血管内皮细胞（BMEC）。 VIM已成为许多参与细胞粘附/迁移和细胞信号传导的许多关键蛋白质的组织者。 IBEA诱导的NF-KB激活，病原体渗透和PMNT在BBB上是必需的VIM和PSF介导的信号传导，这是细菌脑膜炎的三个标志性特征。脂质筏（LRS）用作VIM，PSF，7 NACHR（炎症调节剂）和其他信号分子（例如Caveolin-1，ERK和CAM激酶II）的信号平台。我们最近的研究首次表明VIM和PSF合作调节了IBEA+大肠杆菌诱导的NF-KB信号传导。 VIM通过其头域（HD）在静息细胞中与IKB，NF-KB和小管形成复合物。泛素蛋白酶体系统（UPS）的VIM介导的调节对于NF-KB激活至关重要。 IBEA能够诱导IKB激酶（IKK）激活，VIM/IKB-NF-KB复合物的解离以及VIM的表达，€7 NACHR和促炎因子（PIFS）。 NF-KB的核转运与核中PSF的增加相关。血液中循环的BMEC（CBMEC）和内皮祖细胞（EPC）可用作基于细胞的生物标志物，用于索引中枢神经系统损伤并监测NSM期间基因组范围的表达特征（GES）。最重要的是，我们已经表明，IBEA+ E. coli诱导的NF-KB激活，细菌侵袭，PMNT和BBB渗透性在VIM KO（ - / - ）小鼠中显着降低，这与体外发现一致。基于这些结果，我们假设VIM和PSF诱导的信号传导是一种新的范式，其中微生物因子（例如IBEA）诱导NF-KB的细胞质激活和通过与其他信号分子的初始信号复合物形成NF-KB的核转运（例如，caveolin-caveolin-caveolin-cam，pi3k，pi3kk，筏。随后的IKK/NF-KB途径的激活以及VIM，7 NACHR和PIFS的事件上调导致CNS注入/BBB疾病，可以通过基于CBMEC/EPC的方法检测到。我们的假设将以以下特定目的进行检查：1）。检查VIM和PSF如何协同作用以调节IBEA诱导的NF-KB激活/易位以及NF-KB随后如何通过定义VIM和PSF介导的信号传导来上调基因表达。 2）。通过SCP分析CBMEC和VIM的遗传阻塞，确定VIM和PSF如何促进IBEA诱导的病原体渗透和PMN跨BBB的传播。