A novel proteolytic system of pulmonary inflammation

肺部炎症的新型蛋白水解系统

基本信息

批准号：
8101081
负责人：
AMIT GAGGAR
金额：
$ 36.63万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-02 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8101081
关键词：
Address Adult Animals Antibodies Applications Grants Automobile Driving Biological Markers CXC Chemokines Chemotaxis Chronic Clinical Collagen Cystic Fibrosis Detection Development Disease Dose Epithelial Exhibits Extracellular Matrix Future Generations Glycine Hereditary Disease Hospitalization In Vitro Incubated Individual Inflammation Inflammatory Inflammatory Response Inpatients Interleukin-8 Laboratories Lead Ligands Lung diseases Mass Spectrum Analysis Matrix Metalloproteinases Mediating Metalloproteases Modeling Mus N-terminal Neonatal Pathogenesis Pathologic Pathway interactions Patient Recruitments Patients Peptide Hydrolases Peptides Peripheral Phenotype Play Pneumonia Process Production Proline Protocols documentation Pulmonary Cystic Fibrosis Reaction Reagent Regulation Relative (related person)Research Role Sampling Series Serum Signal Pathway Signal Transduction Sodium Channel Specificity Specimen Sputum Stimulus System Testing Work aerosolized airway inflammation airway remodeling cofactor cystic fibrosis airway cystic fibrosis patients epithelial Na+ channel feeding in vivo mouse model neutrophil novel patient population prolyl oligopeptidase public health relevance receptor binding therapeutic target

项目摘要

DESCRIPTION (provided by applicant): In this proposal, we describe a new pathway signaling neutrophil (PMN) influx and damage to the airways that may play a role as an initiator or cofactor for chronic inflammatory lung diseases such as cystic fibrosis (CF). We have previously demonstrated that proline-glycine-proline (PGP) peptides can cause a robust PMN influx into the airways of mice; this cellular specificity is due to structural relatedness to a receptor binding domain of CXC chemokines such as IL-8. The PI has shown that PGP is released from collagen by a stepwise process initially involving matrix metalloproteases (MMP)-8 and/or 9 with prolyl endopeptidase (PE) catalyzing the final reaction. Recently, the PI's lab has shown that PE is found in neutrophils and stimulated PMNs, when incubated on collagen, can cause notable PGP generation. Although work from the PI's group has demonstrated that PGP is found in CF airway secretions, the impact of PGP peptides in augmenting inflammatory responses in CF lung disease is not currently known. This grant application aims to address this by first investigating the impact of CXC ligands (including PGP and N-(-PGP) as a novel mechanism for the generation of PGP in ex vivo studies and further examining the specific intracellular pathways utilized for protease release from neutrophils. Then, these observations will be taken to a unique mouse model of CF lung disease, the (-ENaC overexpressor mouse (Specific Aim 2), to determine the impact of components of this inflammatory pathway on the phenotype and inflammatory response observed in this murine model. Determination of these fundamental pathways driving protease regulation/release have important implications to patients with chronic inflammatory disease, such as CF, where inflammatory stimuli such as CXC chemokines, are in increased abundance even during periods of disease stability. Our preliminary evidence suggests that at the beginning of an inpatient CF exacerbation, PGP levels are elevated but do decline by the end of hospitalization. However, even with this clinical improvement, PGP levels are still elevated compared to non-disease controls, suggesting that there continues to be ongoing inflammation in these airways. In the final aim of this proposal, we examine if clinically stable CF individuals have elevated levels of PGP and proteases in both sputum and serum. If so, it is possible that their elevation and persistence may predict the development of future disease exacerbations, serving as a unique biomarker for CF lung disease. The findings generated from these aims may have profound translational implications not only to CF lung disease but potentially to other neutrophilic inflammatory conditions. PUBLIC HEALTH RELEVANCE: We have proposed a series of complementary experimental aims to examine the impact of PGP-containing peptides and their generating proteases in cystic fibrosis (CF). The implications of these findings may be broadly applicable to other conditions with prominent neutrophilic inflammation and extracellular matrix breakdown.

描述（由申请人提供）：在本提案中，我们描述了一种新的中性粒细胞（PMN）流入和气道损伤信号通路，该通路可能作为慢性炎症性肺病（如囊性纤维化（CF））的引发剂或辅助因子发挥作用。我们之前已经证明，脯氨酸-甘氨酸-脯氨酸（PGP）肽可以导致大量中性粒细胞流入小鼠气道；这种细胞特异性是由于与 CXC 趋化因子（例如 IL-8）的受体结合域的结构相关性所致。 PI 表明，PGP 通过逐步过程从胶原蛋白中释放，该过程最初涉及基质金属蛋白酶 (MMP)-8 和/或 9，以及催化最终反应的脯氨酰内肽酶 (PE)。最近，PI 的实验室表明，PE 存在于中性粒细胞中，并且当与胶原蛋白一起孵育时，受刺激的 PMN 会导致显着的 PGP 生成。尽管 PI 小组的工作已证明在 CF 气道分泌物中发现了 PGP，但目前尚不清楚 PGP 肽对增强 CF 肺部疾病炎症反应的影响。本拨款申请旨在通过首先研究 CXC 配体（包括 PGP 和 N-(-PGP)）作为在离体研究中生成 PGP 的新机制的影响，并进一步研究用于蛋白酶释放的特定细胞内途径来解决这一问题。然后，将这些观察结果应用于 CF 肺部疾病的独特小鼠模型（-ENaC 过度表达小鼠（具体目标 2）），以确定该炎症途径的成分对在该小鼠模型中观察到的表型和炎症反应对于患有慢性炎症性疾病（例如 CF）的患者具有重要意义，其中 CXC 趋化因子等炎症刺激物即使在慢性炎症期间也会增加。我们的初步证据表明，在住院 CF 恶化开始时，PGP 水平升高，但在住院结束时下降。然而，即使有了这种临床改善，与非疾病对照相比，PGP 水平仍然升高，表明这些气道中仍然存在持续的炎症。在该提案的最终目标中，我们检查临床稳定的 CF 个体的痰液和血清中 PGP 和蛋白酶水平是否升高。如果是这样，它们的升高和持续存在可能可以预测未来疾病恶化的发展，作为 CF 肺病的独特生物标志物。这些目标产生的发现可能不仅对 CF 肺病产生深远的转化意义，而且可能对其他中性粒细胞炎症性疾病产生深远的影响。公共健康相关性：我们提出了一系列补充实验目的，以检查含 PGP 的肽及其生成的蛋白酶对囊性纤维化 (CF) 的影响。这些发现的意义可能广泛适用于具有显着中性粒细胞炎症和细胞外基质分解的其他病症。