ANALYSIS OF AIRWAY SERPINS IN BABOONS MODELS OF BPD

狒狒BPD模型中气道SERPIN的分析

基本信息

批准号：
7716069
负责人：
SULE CATALTEPE
金额：
$ 2.26万
依托单位：
TEXAS BIOMEDICAL RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7716069
关键词：
Apoptosis Aspirate substance Biochemical Biological Assay Blood capillaries Bronchopulmonary Dysplasia Cells Clinical Co-Immunoprecipitations Computer Retrieval of Information on Scientific Projects Database Cysteine Protease Development Endopeptidases Endothelial Cells Epithelial Cells Extracellular Matrix Family Funding Grant Histopathology Immunoblotting Immunohistochemistry Infant Inflammation Inflammatory Institution Kinetics Liquid substance Localized Lung Mass Fragmentography Messenger RNA Modeling Molecular Monitor Morbidity - disease rate Natural Immunity Neonatal Intensive Care Ovalbumin Papio Pathologic Processes Peptide Hydrolases Physiological Play Predisposition Preventive Property Protease Inhibitor Proteins Recombinants Regulation Research Research Personnel Resources Reverse Transcriptase Polymerase Chain Reaction Role Serine Serpins Source Structure of parenchyma of lung Therapeutic United States National Institutes of Health airway inflammation angiogenesis base capillary cell type in vivo lung injury macrophage member mortality nano neutrophil novel protein expression

项目摘要

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. Despite recent advances in neonatal intensive care, bronchopulmonary dysplasia (BPD) remains a major cause of infant morbidity and mortality. The mechanisms for the development of BPD are multifactorial and not yet clearly defined. Inflammation is a common feature of BPD. Airway inflammation is associated with an influx of inflammatory cells and their products. Neutrophils, macrophages and airway epithelial cells release potent proteinases during the inflammatory cascade. These proteinases are involved in diverse physiologic and pathologic processes, such as extracellular matrix remodeling, angiogenesis, apoptosis and innate immunity. In the healthy lung as well as during normal development, the activity of proteinases are tightly regulated by local and systemic anti-proteinases. Among the local proteinase inhibitors, members of the ov-serpin family (ovalbumin-related serpins) are an emerging group of proteins that are abundantly expressed by several cell types in the lung, including airway epithelial cells, endothelial cells and inflammatory cells. These ov-serpins include SERPINB1, -B2, -B3, -B4, B6 and B9. Ov-serpins inhibit an array of proteinases that play significant roles in lung injury. Based on their localization, regulation and biochemical properties, we hypothesize that ov-serpins are ideally localized in the lung tissue and inflammatory cells to regulate the activity of proteinases released during inflammation, such as occurs in BPD. Transcriptional or post-translational alterations in ov-serpin expression in the immature lung can be associated with unopposed proteinase activity and thus, increased susceptibility to BPD. In order to investigate our hypothesis, we propose to utilize the well-characterized baboon models of BPD. The specific aims of this project are to: 1) correlate ov-serpin mRNA and protein expression in baboon lungs with the development of BPD by quantitative RT-PCR, immunoblotting, and immunohistochemistry, 2) characterize the activity of serine- and cysteine proteinases as ov-serpin targets in the airways of baboons with and without BPD by kinetic assays and identify in vivo target proteinases of ov-serpins by analyzing tracheal aspirate fluids by co-immunoprecipitation and nano-capillary HPLC-ion trap mass spectrometry (LC-MS/MS), 3) determine whether administration of recombinant SCCA1 (SERPINB3) alters the development of BPD in baboon models by monitoring clinical and biochemical parameters, and lung histopathology. These studies will enhance our understanding of the cellular and molecular mechanisms underlying BPD and facilitate development of novel preventive and therapeutic strategies for BPD.

该子项目是利用该技术的众多研究子项目之一资源由 NIH/NCRR 资助的中心拨款提供。子项目和研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金，因此可以在其他 CRISP 条目中表示。列出的机构是对于中心来说，它不一定是研究者的机构。尽管新生儿重症监护最近取得了进展，但支气管肺发育不良（BPD）仍然是婴儿发病和死亡的主要原因。 BPD 的发生机制是多因素的，目前尚未明确定义。炎症是 BPD 的一个常见特征。气道炎症与炎症细胞及其产物的流入有关。中性粒细胞、巨噬细胞和气道上皮细胞在炎症级联反应期间释放有效的蛋白酶。这些蛋白酶参与多种生理和病理过程，例如细胞外基质重塑、血管生成、细胞凋亡和先天免疫。在健康的肺部以及正常发育过程中，蛋白酶的活性受到局部和全身抗蛋白酶的严格调节。在局部蛋白酶抑制剂中，ov-丝氨酸蛋白酶抑制剂家族（卵清蛋白相关丝氨酸蛋白酶抑制剂）的成员是一组新兴蛋白质，在肺部多种细胞类型中大量表达，包括气道上皮细胞、内皮细胞和炎症细胞。这些 ov-丝氨酸蛋白酶抑制剂包括 SERPINB1、-B2、-B3、-B4、B6 和 B9。 Ov-丝氨酸蛋白酶抑制剂可抑制一系列在肺损伤中发挥重要作用的蛋白酶。基于它们的定位、调节和生化特性，我们假设 ov-丝氨酸蛋白酶抑制剂理想地定位于肺组织和炎症细胞中，以调节炎症期间释放的蛋白酶的活性，例如 BPD 中发生的情况。未成熟肺中 ov-serpin 表达的转录或翻译后改变可能与不受对抗的蛋白酶活性相关，因此增加了对 BPD 的易感性。为了研究我们的假设，我们建议利用特征良好的 BPD 狒狒模型。该项目的具体目标是：1) 通过定量 RT-PCR、免疫印迹和免疫组织化学将狒狒肺中的 ov-serpin mRNA 和蛋白表达与 BPD 的发展相关联，2) 将丝氨酸和半胱氨酸蛋白酶的活性表征为通过动力学测定，ov-serpin 靶向有或没有 BPD 的狒狒气道，并通过分析识别 ov-serpin 的体内靶蛋白酶通过免疫共沉淀和纳米毛细管 HPLC-离子阱质谱 (LC-MS/MS) 分析气管抽吸液，3) 通过监测临床和生化参数确定重组 SCCA1 (SERPINB3) 的施用是否会改变狒狒模型中 BPD 的发展和肺组织病理学。这些研究将增强我们对 BPD 背后的细胞和分子机制的理解，并促进开发新的 BPD 预防和治疗策略。