Viral and Genetic Regulation of Abnormal OCL Activity in PD

PD 中异常 OCL 活性的病毒和遗传调控

• 批准号: 8315745
• 负责人: DEBORAH Lynn GALSON
• 金额: $ 31.54万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-16 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8315745
• 关键词: Animal Model; CCAAT-Enhancer-Binding Proteins; CREB1 gene; Calcineurin; Cells; Complex; Cytokine Signaling; Funding; Genes; Genetic Transcription; Grant; IKKepsilon; IRF3 gene; Interleukin-6; Link; MAP Kinase Gene; MAPK14 gene; Measles virus nucleoprotein; Mediating; Messenger RNA; Molecular; Mutation; NF-kappa B; Osteoclasts; Paget' s Disease; Pathogenesis; Pathway interactions; Phenotype; Phosphorylation; Phosphotransferases; Proteins; Regulation; Reporting; Research Personnel; Role; Scaffolding Protein; Serine; Signal Transduction; TANK-binding kinase 1; TNF gene; TNFSF11 gene; TRAF2 gene; Transcription Factor AP-1; Tumor Necrosis Factor-alpha; Up-Regulation; Vitamin D3 Receptor; activating transcription factor; cofactor; cytokine; factor C; human TNF protein; in vivo; insight; novel; osteoclastogenesis; p65; programs; protein expression; response; transcription factor; virus genetics

Both viral and genetic factors have been implicated in the pathogenesis of Paget's disease (PD). However, the molecular mechanisms responsible for their effect on osteoclast (OCL) formation and activity in PD are unclear. It is our hypothesis that measles virus nucleoprotein (MVNP) mediates its effects in OCL precursors predominantly through increased NFicB signaling via activation of the two kB kinase (IKK)-related kinases IKKe (also called IKK-/) and TANK-binding kinase 1 (TBK1) and increased levels of RIP1 and TRAF2. This results in upregulation of IL-6 and a vitamin D receptor coactivator TAFM-17 and increases OCL formation. MVNP also drives multi-potential cells towards the OCL lineage through increased expression and activation of NFATd. The p62P392L mutation linked to PD enhances the effects of MVNP by further increasing NFxB signaling in response to TNF-ct and RANKL as well as increasing p38 MAPK activation in response to RANKL, TNF-alpha and 1,25-(OH)2D3. In support of this hypothesis, we recently found that MVNP expression increases RIP1, TRAF2, and p65 NFicB resulting in enhanced constitutive activation of NFicB, and increased NFicB responsivity to TNF-alpha and RANKL. We also found that MVNP increases NFATd and synergistically activates NFATcl-driven gene transcription, with either NFATd 'or RANKL, and induces IL-6 and TAFn-17. In addition, the p62p392L mutation activates NFicB and increases the sensitivity of OCL precursors to RANKL and TNF-alpha. MVNP also interacts with IKKe and TBK1 to activate NFKB but their role in PD are unknown. In this project we will: 1. Determine if MVNP activation of NFKB and induction of pagetic OCL formation results from alterations of the canonical pathway and/or utilization of an alternative pathway that involves activation of IKKepsilon/TBK1 and the effects of co-expression of MVNP and p62P392L on these pathways. 2. Determine if NFKB activation by MVNP is sufficient for induction of IL-6 or if other mechanisms are also necessary, and the effects of co-expression of p62R392L and MVNP on the molecular mechanlsm(s) identified above. 3. Determine if the synergy of MVNP with NFATd is due to modulation of NFATd activation or of a cofactor of NFATd. The studies of the molecular mechanisms responsible for MVNP and p62p392L effects on OCL formation outlined above" will provide important insight into the pathogenesis of PD and are directly linked to the cellular and in vivo studies proposed in Project 2,

佩吉特病（PD）的发病机制与病毒和遗传因素有关。然而，负责其对破骨细胞（OCL）的形成和活性在PD的影响的分子机制尚不清楚。我们假设麻疹病毒核蛋白（MVNP）主要通过激活两种kB激酶（IKK）相关激酶IKKe（也称为IKK-/）和TANK结合激酶1（TBK 1）增加NFicB信号传导以及增加RIP 1和TRAF 2水平来介导其在OCL前体中的作用。这导致IL-6和维生素D受体共激活剂TAFM-17的上调，并增加OCL的形成。MVNP还通过增加NFATd的表达和活化驱动多潜能细胞朝向OCL谱系。与PD相关的p62 P392 L突变通过响应于TNF-α和RANKL进一步增加NFxB信号传导以及响应于RANKL、TNF-α和1，25-（OH）2D 3增加p38 MAPK活化来增强MVNP的作用。为了支持这一假设，我们最近发现MVNP表达增加RIP 1、TRAF 2和p65 NFicB，导致NFicB的组成性激活增强，并增加NFicB对TNF-α和RANKL的响应性。我们还发现，MVNP增加NFATd，并协同激活NFATc 1-驱动的基因转录，与NFATd '或RANKL，并诱导IL-6和TAFn-17。此外，p62 p392 L突变激活NFicB并增加OCL前体对RANKL和TNF-α的敏感性。MVNP还与IKKe和TBK 1相互作用以激活NF κ B，但它们在PD中的作用尚不清楚。在这个项目中，我们将：1。确定NF κ B的MVNP激活和pagetic OCL形成的诱导是否是由于经典途径的改变和/或涉及IKK β/TBK 1激活的替代途径的利用以及MVNP和p62 P392 L共表达对这些途径的影响。2.确定MVNP对NF κ B的激活是否足以诱导IL-6或是否还需要其他机制，以及p62 R392 L和MVNP的共表达对上述分子机制的影响。3.确定MVNP与NFATd的协同作用是否是由于NFATd活化或NFATd辅因子的调节。上述负责MVNP和p62 p392 L对OCL形成的影响的分子机制的研究”将为PD的发病机制提供重要的见解，并与项目2中提出的细胞和体内研究直接相关，