A novel proteolytic system of pulmonary inflammation

肺部炎症的新型蛋白水解系统

• 批准号: 8475396
• 负责人: AMIT GAGGAR
• 金额: $ 34.65万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 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/8475396
• 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

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.

描述（由申请人提供）：在此提案中，我们描述了一种新的途径信号中性粒细胞（PMN）涌入和对气道的损害，可能在慢性炎性肺部疾病（例如囊性纤维化（CF））中起发挥作用。我们以前已经证明，脯氨酸 - 甘氨酸 - 丙啉（PGP）肽会导致小鼠气道的强大PMN涌入。该细胞特异性是由于与CXC趋化因子（例如IL-8）的受体结合结构域的结构相关性。 PI表明，PGP通过最初涉及金属蛋白酶（MMP）-8和/或9的逐步过程从胶原蛋白中释放出来，并用prolyl内肽酶（PE）催化了最终反应。最近，PI的实验室表明，在嗜中性粒细胞中发现了PE，并且在胶原蛋白孵育时刺激的PMN可能会引起显着的PGP产生。尽管PI小组的工作表明PGP在CF气道分泌物中发现，但PGP肽对增加CF肺部疾病的炎症反应的影响尚不清楚。该赠款申请旨在通过首先研究CXC配体的影响（包括PGP和N - （ - PGP），作为离体研究中PGP生成的新机制，并进一步研究用于从蛋白酶释放的特定细胞内途径中性粒细胞。这些基本途径的鼠模型都对慢性炎症性疾病的患者（例如CF）具有重要意义。在住院CF加剧开始时，PGP水平升高，但在住院结束时确实下降了。但是，即使有了这种临床改善，与非疾病控制相比，PGP水平仍然升高，这表明这些气道中仍在持续炎症。在该提案的最终目的中，我们检查了临床稳定的CF个体在痰液和血清中的PGP和蛋白酶水平是否升高。如果是这样，他们的升高和持久性可能会预测未来疾病加剧的发展，这是CF肺部疾病的独特生物标志物。这些目标产生的发现可能不仅对CF肺疾病具有深远的翻译意义，而且可能对其他中性粒细胞炎症状况具有深远的影响。