NONINVASIVE BIOMARKERS OF PROTEOLYTIC ACTIVITY IN CHILDREN WITH CYSTIC FIBROSIS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 在患有囊性纤维化(CF)的儿童中，蛋白分解活性会导致支气管扩张，导致进行性肺部疾病和显著的预期寿命缩短。这项建议的长期目标之一是定义预测儿童CF未来临床病程和疾病进展的蛋白水解性生物标志物。通过识别那些蛋白分解活性过高和更具侵略性的儿童，有可能在不可逆转的呼吸道损伤发生之前进行抗蛋白分解治疗。主要的假设是，在诱导痰中检测到有更明显的蛋白分解活性的CF儿童，将有更大程度的结构性和功能性肺损伤。这一假说将通过以下具体目标来验证：1)通过定量测定临床稳定期的CF患儿临床标本(诱导痰、尿液)中性粒细胞衍生的蛋白水解酶(弹性蛋白酶、基质金属蛋白酶2和9型)、肺抗蛋白水解酶(α-抗蛋白水解酶、分泌型白蛋白水解酶抑制物、金属蛋白酶组织抑制物)和弹性蛋白分解产物(桥桥素、异桥桥素)的水平，以确定蛋白分解活性的变化；2)将这些蛋白分解活性的变化与呼吸道结构性损害(通过每年高分辨率计算机断层扫描的支扩的严重程度和程度以及通过测量痰和尿中弹性蛋白分解产物进行生化评估)、功能性呼吸道损害(由年度肺功能测试确定)、较低的呼吸道细菌定植状况和细菌负荷以及相关发病率(住院率、肺部恶化)相关联；以及3)通过确定蛋白酶水平是否与被认为改变CF肺部疾病的基因(MBL、肿瘤坏死因子、转化生长因子-1和A1AT)的多态性有关，来检测基因修饰物对气道蛋白分解活性的影响。这些结果将是评估儿童CF新出现的抗蛋白水解性治疗的关键。 这项临床研究方案的另一个目标是加强和加强萨格尔博士在临床研究和以患者为中心的研究方面的方法。萨格尔博士将通过完成科罗拉多大学临床科学项目的博士学位，接受更正规的培训和教育。他将学习临床流行病学、生物伦理学、临床试验设计、药代动力学和人类遗传学等课程，并完成一篇关于CF中蛋白质分解活性的论文。此外，他将积极参与和培训儿科GCRC，并经常与他的赞助商、导师和合作者互动。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In children with cystic fibrosis (CF), proteolytic activity causes bronchiectasis, resulting in progressive lung disease and marked shortening of life expectancy. One of the long term objectives for this proposal is to define proteolytic biomarkers that are predictive of future clinical course and disease progression in children with CF. By identifying those children with excessive and more aggressive proteolytic activity, it may be possible to intervene with anti-proteolytic treatments before irreversible airway damage occurs. The main hypothesis is that CF children with more pronounced proteolytic activity, as measured in induced sputum, will have a greater degree of structural and functional lung damage. This hypothesis will be tested through the following specific aims: 1) to determine changes in proteolytic activity by quantitating levels of neutrophil-derived proteases (elastase, matrix metalloproteinases types 2 and 9), lung antiproteases (alpha1-antiprotease, secretory leukoprotease inhibitor, tissue inhibitors of metalloproteinases), and elastin breakdown products (desmosine, isodesmosine) in clinical specimens (induced sputum, urine) from CF children, during times of clinical stability, annually over 3 years; 2) to correlate these changes in proteolytic activity with structural airway damage (assessed by severity and extent of bronchiectasis on annual high-resolution computed tomography scans as well as biochemically through measurement of elastin breakdown products in sputum and urine), functional airway impairment (as determined by annual pulmonary function testing), lower airway bacterial colonization status and bacterial burden, and related morbidities (rates of hospitalization, pulmonary exacerbations); and 3) to examine the influence of genetic modifiers on airway proteolytic activity by determining if protease levels are associated with polymorphisms in genes that are believed to modify CF lung disease (MBL, TNF , TGF- , and A1AT). These results will be crucial to evaluating emerging anti-proteolytic treatments in children with CF. Another objective of this clinical research protocol is to enhance and strengthen Dr. Sagel's approach to clinical investigation and patient-oriented research. Dr. Sagel will receive more formal training and education by completing his Ph.D. in the University of Colorado's Clinical Science Program. He will take courses in clinical epidemiology, bioethics, clinical trial design, pharmacokinetics, and human genetics, and complete a thesis about proteolytic activity in CF. In addition, he will actively participate and train in the Pediatric GCRC, and frequently interact with his sponsor, mentors, and collaborators.