Cardiolipin Induced Lung Injury and FIC1

心磷脂引起的肺损伤和 FIC1

基本信息

批准号：
7799007
负责人：
Rama K Mallampalli
金额：
$ 49.3万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
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：
2010-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7799007
关键词：
Acute Acute Lung Injury Adverse effects Alveolar Alveolar Cell Apical Apoptosis Apoptotic Aspirate substance Attenuated Bacterial Infections Binding Biochemical Biological Biological Assay Bronchoalveolar Lavage Cardiolipins Carrier Proteins Cartoons Cell Line Cells Chemicals Chronic Chronic Obstructive Airway Disease Complement Coupled Data Disease Distal Dominant-Negative Mutation Epithelial Epithelial Cells Epithelium Escherichia coli Escherichia coli Infections Exhibits Foundations Gene Transfer Generations Human Impairment In Vitro Infection Inflammation Mediators Inflammatory Irrigation Knock-out Laboratories Lead Left Link Lipids Liquid substance Lung Lung Compliance Lung Lavage Fluid Lung diseases Maps Mechanics Mediating Mediator of activation protein Membrane Metabolism Mitochondria Modeling Molecular Molecular Profiling Mus Mutant Strains Mice Pathway interactions Patients Peptides Pharmaceutical Preparations Phosphatidylserines Phospholipids Physiological Physiological Processes Physiology Pneumonia Process Property Proteins Pseudomonas aeruginosa Pump RNA Interference Regulation Respiratory physiology Role Sepsis Severity of illness Structure Surface Tension Teaching Hospitals Testing Time Transgenic Mice Ubiquitin Universities Ventilator Virulent Work alveolar epithelium base clinically relevant human disease in vitro activity in vivo inhibitor/antagonist lung injury member microbial novel novel strategies novel therapeutics overexpression pathogen patient oriented programs protein expression public health relevance response sensor small molecule surfactant targeted delivery tool trafficking ubiquitin-protein ligase uptake

项目摘要

DESCRIPTION (provided by applicant): Cardiolipin (CL) is an important phospholipid component of mitochondrial and bacterial membranes that has emerged as a critical factor in the apoptotic program. To date, there is very limited data on the metabolism, processing, and physiologic role of CL in human disease states. Normally, very little CL is detected in lung fluid or in association with surfactant, but levels increase in patients with chronic obstructive pulmonary disease (COPD), and in chemical and sepsis-induced lung injury models. Preliminary data in the PI's laboratory shows that CL is elevated in lung fluid of patients with ventilator- associated pneumonia and in murine models of pneumonitis. Further, CL potently elevates lung surface- tension and impairs lung compliance. These original observations suggest a new role for CL as an important mediator of acute and chronic inflammatory lung injury. The data also suggest the existence of a CL-alveolar transport protein that may evacuate this phospholipid from pulmonary fluid, thereby preserving lung structure and function. ATP8b1 is a relatively newly discovered lipid pump that functions to translocate a phospholipid, phosphatidylserine, across biological membranes. Preliminary data show that ATP8b1 is highly expressed in alveolar epithelia, that ATP8b1 binds CL, and stable overexpression of this pump increases uptake and internalization of CL in murine lung epithelia. ATP8b1 mutant mice exhibit impairment of lung biophysical properties and are severely prone to acute bacterial inflammatory lung injury. Collectively, the above observations led to the overall hypothesis in this project that cardiolipin is a key mediator of inflammatory lung injury, and its intra-alveolar availability is tightly regulated by the lipid pump, ATP8b1. The PI will conditionally express ATP8b1 in a cell line and generate distal lung epithelial-targeted ATP8b1 double transgenic mice to ascertain the role of this lipid pump in regulating CL availability in lung fluid after E. coli and P. aeruginosa infection. Studies also assess ATP8b1 protein apical targeting, delivery of ATP8b1 CL-binding decoy peptides, and gene transfer of ATP8b1 in mutant mice after E. coli and P. aeruginosa infection as a means to attenuate inflammatory lung injury. Last, studies will assay CL levels and molecular species of CL in patients with acute lung injury at a University-based, patient-oriented teaching hospital. Execution of these studies will provide novel information of ATP8b1 molecular regulation that will impact the field of lung inflammatory disease. PUBLIC HEALTH RELEVANCE: We have recently discovered that in pneumonia, there is marked elevation of a rare lipid, termed cardiolipin, that potently inhibits lung function. We have also discovered a lipid pump, termed FIC1, that is rapidly degraded after bacterial infection. We propose in this application to use several tools to confirm that FIC1 is indispensable for controlling the availability of cardiolipin in lung fluid, to identify the molecular and biochemical mechanisms for FIC1 breakdown after bacterial infection, and to correlate cardiolipin levels in lung fluid of patients having pneumonia with illness severity.

描述（由申请人提供）：Cardiolipin（Cl）是线粒体和细菌膜的重要磷脂成分，它已成为凋亡程序的关键因素。迄今为止，CL在人类疾病状态中的代谢，加工和生理作用的数据非常有限。通常，在肺液或与表面活性剂相关的情况下检测到很少的CL，但是慢性阻塞性肺疾病（COPD）以及化学和败血症诱导的肺损伤模型的患者的水平增加。 PI实验室中的初步数据表明，呼吸机相关肺炎患者和肺炎鼠模型中的肺液中CL升高。此外，CL可有效提高肺表面张力并损害肺部依从性。这些原始观察结果表明，CL是急性和慢性炎症肺损伤的重要介质。数据还表明存在可从肺液中撤离这种磷脂的Cl-肺泡转运蛋白，从而保存肺结构和功能。 ATP8B1是一种相对较新的脂质泵，可在生物膜跨生物膜中运输磷脂，磷脂酰丝氨酸。初步数据表明，ATP8B1在肺泡上皮中高度表达，ATP8B1结合Cl，并且该泵的稳定过表达增加了鼠肺上皮中Cl的摄取和内在化。 ATP8B1突变小鼠表现出肺部生物物理特性损害，并且严重容易发生急性细菌炎症性肺损伤。总体而言，上述观察结果导致该项目的总体假设是心磷脂是炎症性肺损伤的关键介体，其肺泡内的可用性受脂质泵ATP8B1严格调节。 PI将有条件地在细胞系中表达ATP8B1，并产生远端的肺上皮靶向ATP8B1双重转基因小鼠，以确定该脂质泵在大肠杆菌和P. aeruginosa感染后调节肺液中CL的可用性中的作用。研究还评估了ATP8B1蛋白根尖靶向，ATP8B1 CL结合诱饵肽的递送以及大肠杆菌和铜绿假单胞菌感染后突变小鼠中ATP8B1的基因转移，以此作为减轻炎症性肺损伤的一种手段。最后，研究将在急性肺损伤患者的一家大学，以患者为导向的教学医院的急性肺损伤患者中分析CL水平和分子种类。这些研究的执行将提供ATP8B1分子调节的新信息，这将影响肺部炎症性疾病的领域。公共卫生相关性：我们最近发现，在肺炎中，一种稀有脂质（称为心磷脂）的明显升高，可有效抑制肺功能。我们还发现了一个称为FIC1的脂质泵，该脂质在细菌感染后迅速降解。我们建议在此应用中使用多种工具来确认FIC1对于控制肺液中有心脂蛋白的可用性是必不可少的，以识别细菌感染后FIC1分解的分子和生化机制，以及相关肺炎患病严重程度。