The Role of ARNO-Arf6 signaling in barrier stability and chronic rhinosinusitis

ARNO-Arf6 信号在屏障稳定性和慢性鼻窦炎中的作用

• 批准号: 9060254
• 负责人: ANDREW P LANE
• 金额: $ 20.25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9060254
• 关键词: ADP-Ribosylation Factors; ADP-ribosylation factor 6; Affect; Antigens; Arthritis; Biochemistry; Biological Models; Blood Vessels; Cell Culture Techniques; Cell Surface Proteins; Cells; Cellular biology; Chronic; Chronic Sinusitis; Complex; Cytokine Signaling; Disease; Dyes; Endothelial Cells; Endothelium; Epithelial; Epithelial Cell Junction; Epithelial Cells; Equilibrium; Etiology; Evans blue stain; Functional disorder; Genes; Genetic; Guanine Nucleotide Exchange Factors; Health; Human; Immune; Immunology; Infiltration; Inflammation; Inflammatory; Intercellular Junctions; Interleukin-1 beta; Knock-out; Knockout Mice; Leukocytes; Liquid substance; Measures; Mediating; Medical; Modeling; Molecular; Mucous Membrane; Mus; NF-kappa B; Nasal Polyps; Nature; Nucleotides; Operative Surgical Procedures; Ovalbumin; Pathologic; Pathology; Pathway interactions; Patients; Permeability; Play; Polyps; Principal Investigator; Proteins; Publishing; Regulation; Research; Role; Signal Pathway; Signal Transduction; Sinus; Sinusitis; Small Interfering RNA; Stimulus; System; Techniques; Testing; Tissues; Transgenic Mice; United States; Vascular Permeabilities; Vascular System; Work; cell type; chronic rhinosinusitis; cytokine; drug development; improved; in vivo; indexing; innate immune function; mouse model; programs; research study; rhinosinusitis; small molecule inhibitor; therapeutic target; trafficking

 DESCRIPTION (provided by applicant): Chronic rhinosinusitis (CRS) is a significant health problem in the United States. It has been hypothesized that a common etiologic pathway may involve transient or chronic alterations to the sinonasal epithelial barrier perhaps allowing passage of agents that perpetuate a cycle of antigen exposure, increased cytokine release, and inflammation. There are two histopathologic subtypes of CRS, one without and one with nasal polyps (CRSsNP and CRSwNP respectively). The cellular and molecular mechanisms that underlie CRNsNP and CRSwNP remain poorly understood. A recent study published in Nature from the lab of our collaborator Dr. Dean Li demonstrated that the inflammatory cytokine interleukin-1 beta (IL-1ß) activates the ADP ribosylation factor nucleotide exchange factor (ARNO)-ADP-ribosylation factor 6 (Arf6) pathway independently of NF-κB to reduce inter-cellular junction protein cell surface localization in the vasculature. Our preliminary studies suggest that one can enhance sinonasal epithelial barrier stability and reduce indicies of CRS through targeted inhibition of the ARNO-Arf6 pathway. We hypothesize that as previously demonstrated in the vascular system, Arf6 is a central regulator of cytokine-induced destabilization of sinonasal epithelial junction protein cell surface localization and barrier permeability. We hypothesize that the sinonasal epithelial cell barrier in CRSsNP and CRSwNP patients is destabilized at baseline as well as through inflammatory cytokine signaling via increased Arf6 activity, and that inhibition of ARNO-Arf6 signaling will restore stability in both settings in an NF-κB-independent manner. Lastly, we hypothesize that inhibition of ARNO-Arf6 signaling will increase barrier stability and will reduce indicies of CRS. To test these hypotheses we will initially, in aim 1, examine sinus mucosa and characterize baseline and cytokine-induced differences in SNEC junction destabilization in patients without chronic sinusitis, CRSsNP and CRSwNP. We will also determine whether inhibition of Arf6 activity stabilizes the sinonasal epithelial barrier and reduces cytokine-induced barrier permeability. We will also determine whether inhibition of ARNO-Arf6 acts independently of NF-κB signaling in cytokine-treated SNECs. In aim 2, we will utilize genetically modified mice and the ovalbumin mouse model of CRS to determine whether stabilization of the epithelial or endothelial barrier through genetic deletion of the ARNO-Arf6 pathway reduces sinusitis. Fundamentally the studies proposed here seek to understand the pathologic role that barrier instability and permeability play in CRSsNP and CRSwNP and to evaluate the ARNO-Arf6 pathway as a therapeutic target. As elucidation of the barrier stabilizing effects of inhibition of the ARNO-Arf6 system has largely been limited to stabilizing the vascular barrier, this proposal has potential significance and applicability beyond sinusitis to other conditions affecting epithelial barrier stability.

 描述（由申请人提供）：慢性鼻窦炎（CRS）在美国是一个严重的健康问题。据推测，一个共同的病因学途径可能涉及短暂或慢性改变鼻窦上皮屏障，可能允许通过的代理人，永久的抗原暴露，增加细胞因子释放和炎症的周期。CRS有两种组织病理学亚型，一种无鼻息肉，一种有鼻息肉（分别为CRSsNP和CRSwNP）。CRNsNP和CRSwNP的细胞和分子机制仍然知之甚少。我们的合作者Dean Li博士的实验室最近在Nature上发表的一项研究表明，炎症细胞因子白细胞介素-1 β（IL-1 β）激活ADP核糖基化因子核苷酸交换因子（ARNO）-ADP核糖基化因子6（Arf 6）通路，独立于NF-κB，以减少血管系统中细胞间连接蛋白的细胞表面定位。我们的初步研究表明，通过靶向抑制ARNO-Arf 6通路，可以增强鼻窦上皮屏障的稳定性，减少CRS的诱导。我们假设，如先前在血管系统中所证明的，Arf 6是烟碱诱导的鼻窦上皮连接蛋白细胞表面定位和屏障通透性不稳定的中央调节因子。我们假设CRSsNP和CRSwNP患者的鼻窦上皮细胞屏障在基线时以及通过Arf 6活性增加的炎性细胞因子信号传导而不稳定，并且ARNO-Arf 6信号传导的抑制将以NF-κ B非依赖性方式恢复两种情况下的稳定性。最后，我们假设ARNO-Arf 6信号传导的抑制将增加屏障稳定性并将减少CRS的不确定性。为了检验这些假设，我们将首先在目标1中检查窦粘膜，并表征没有慢性鼻窦炎、CRSsNP和CRSwNP的患者中SNEC连接不稳定的基线和尼古丁诱导的差异。我们还将确定是否抑制Arf 6活性稳定的鼻窦上皮屏障，并减少苦参碱诱导的屏障通透性。我们还将确定ARNO-Arf 6的抑制作用是否独立于NF-κB信号传导在马槟榔碱处理的SNEC中起作用。在目标2中，我们将利用CRS的遗传修饰小鼠和卵清蛋白小鼠模型来确定通过ARNO-Arf 6途径的遗传缺失来稳定上皮或内皮屏障是否减少鼻窦炎。从根本上说，这里提出的研究试图了解屏障不稳定性和渗透性在CRSsNP和CRSwNP中发挥的病理作用，并评估ARNO-Arf 6途径作为治疗靶点。由于ARNO-Arf 6系统的抑制的屏障稳定作用的阐明在很大程度上限于稳定血管屏障，因此该提议具有潜在的意义和适用性， 鼻窦炎影响上皮屏障稳定性的其他条件。