Targeting Underlying Pathophysiological Mechanisms to Develop Novel Therapies for Chronic Obstructive Lung Disease

针对潜在的病理生理机制开发慢性阻塞性肺病的新疗法

基本信息

批准号：
10171619
负责人：
Maliha Zahid
金额：
$ 58.39万
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：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10171619
关键词：
Affect Affinity Alanine Alveolar Amino Acids Apoptotic Area Arginine Attenuated Binding Biodistribution Biological Assay Cardiac Cell Culture Techniques Cell Death Cell Density Cell Nucleus Cell Surface Proteins Cell surface Cells Chronic Chronic Obstructive Airway Disease Cilia Cleaved cell Coughing Cytolysis Disease Disease model Embryo Epithelial Epithelial Cells Esters FDA approved Frequencies Functional disorder Generations Genus Hippocampus Goals Goblet Cells Grant Hair Harvest Heart High Prevalence Hospitalization Human Hyperplasia Image Impairment Incubated Inflammatory Infiltrate Inhalation Injections Intravenous Label Lead Length Ligands Lower Respiratory Tract Infection Lung Measures Mitochondria Mitochondrial DNA Morbidity - disease rate Morphology Mucociliary Clearance Mucous body substance Mus Nasal Epithelium Organ Oxygen Pathway interactions Peptides Peripheral Ploidies Population Positioning Attribute Predisposition Production Proteins Reactive Oxygen Species Recurrence Reduced Glutathione Scanning Serine Shortness of Breath Small Interfering RNA Structure of parenchyma of lung Surface Technology Testing Therapeutic Time Tissues Tracheobronchial Trees Triceps Brachii Muscle Tubulin Work Xenopus airway obstruction bronchial epithelium burden of illness cigarette smoke-induced COPD cilium biogenesis cilium motility cytokine drug development esterase exposure to cigarette smoke fluid flow gamma secretase imaging system improved in vivo in vivo imaging system inhibitor/antagonist interest knock-down mitochondrial dysfunction mortality mutant notch protein novel novel therapeutics pathogen pollutant recurrent infection small molecule smoke-induced COPD thioredoxin peroxidase transcription factor uptake vector

项目摘要

Chronic obstructive pulmonary disease (COPD) is characterized by airflow obstruction, increased mucus production, and predisposition to recurrent lower respiratory tract infections. It affects ~5% of the US population, ranking third as a cause of mortality. This high prevalence and disease chronicity results in frequent hospitalizations, and need for lifelong therapies. In spite of this high disease burden, there have been no new FDA approved therapies in the past two decades. Chronic cigarette smoke exposure, the commonest cause of COPD, leads to increased generation of reactive oxygen species (ROS), decreased mitochondrial ability to handle these ROS, and cell death, with the latter leading to breakdown of alveolar surfaces, blebbing, and decreased pulmonary surface for adequate oxygen exchange. Cilia are tubulin-containing hair-like projections on the cell surface of epithelial cells lining the tracheobronchial tree that beat in a coordinated, metachronal wave to sweep inhaled pollutants and pathogens away from the lungs. In COPD, these cilia are sparse, stunted, and beat with lower frequencies than in healthy lungs leading to poor mucociliary clearance (MCC) and recurrent infections. Mitochondrial dysfunction due to overwhelming ROS production is also associated with defective cilia formation. Our prior work led to identification of a 12-amino acid peptide that we termed Cardiac Targeting Peptide due to its ability to transduce normal mouse heart tissue after peripheral injection. An alanine scan with sequential, single alanine substitutions led to the discovery of two alanine mutants (S7A and R11A with serine at position 7 and arginine at position 11 substituted with alanine) that instead of the heart robustly transduced lung epithelial tissue after a peripheral injection. A key player in ciliogenesis is Notch, a transcription factor that determines pluripotent Club cell fate and differentiation into mucus producing goblet cells at the expense of multi-ciliated cells. We have shown that treatment of reciliating mouse tracheal and human nasal epithelial cell cultures with 2nM DAPT (N-[N-(3,5-Difluorophenacetyl)-L- alanyl]-S-phenylglycine t-butyl ester), a small molecule Notch inhibitor, led to significant increase in degree of ciliogenesis, cilia length, and ciliary beat frequency compared to controls. In this grant, we are proposing to develop these novel lung targeting peptides as vectors to deliver a number of different ROS scavengers (Szeto-Schiller peptide, Mitotempo, reduced glutathione) to lungs of mice with smoke-induced COPD. We are also proposing to utilize DAPT in vivo in these mice to improve ciliary function with the ultimate goal of improving MCC. Our overarching goal is to target novel pathophysiological pathways in COPD by enhancing mitochondrial function and ciliogenesis to improve MCC.

慢性阻塞性肺疾病 (COPD) 的特点是气流阻塞、粘液增多生产和复发性下呼吸道感染的倾向。它影响了美国约 5% 的地区人口，位居第三位死亡原因。这种高患病率和疾病长期性导致频繁住院，需要终生治疗。尽管疾病负担很高，但过去二十年来，FDA 没有批准任何新疗法。慢性香烟烟雾暴露是最常见的慢性阻塞性肺病 (COPD) 的病因，导致活性氧 (ROS) 生成增加、线粒体减少处理这些 ROS 和细胞死亡的能力，后者导致肺泡表面破裂、起泡、并减少肺表面积以进行充分的氧气交换。纤毛是含有微管蛋白的毛发状气管支气管树内壁上皮细胞表面的投射，以协调的方式跳动，异时波将吸入的污染物和病原体从肺部清除。在慢性阻塞性肺病中，这些纤毛是与健康肺部相比，稀疏、发育不良和搏动频率较低，导致粘膜纤毛清除不良（MCC）和反复感染。由于过量的 ROS 产生而导致线粒体功能障碍与纤毛形成缺陷有关。我们之前的工作导致鉴定出一种 12 氨基酸肽，我们将其由于其能够在外周血后转导正常小鼠心脏组织，因此被称为心脏靶向肽。注射。通过连续的、单个丙氨酸取代的丙氨酸扫描发现了两个丙氨酸突变体（S7A 和 R11A，其中第 7 位的丝氨酸和第 11 位的精氨酸被丙氨酸取代）外周注射后，代替心脏强烈转导肺上皮组织。关键人物纤毛发生是Notch，一种决定多能俱乐部细胞命运和分化的转录因子以多纤毛细胞为代价产生粘液的杯状细胞。我们已经表明，安抚治疗使用 2nM DAPT (N-[N-(3,5-二氟苯乙酰)-L- 丙氨酰]-S-苯基甘氨酸叔丁酯），一种小分子Notch抑制剂，导致Notch程度显着增加与对照组相比，纤毛发生、纤毛长度和纤毛跳动频率。在这笔赠款中，我们建议开发这些新型肺靶向肽作为载体来传递许多不同的 ROS 清除剂（Szeto-Schiller 肽、Mitotempo、还原型谷胱甘肽）对烟雾诱发的慢性阻塞性肺病小鼠的肺部。我们是还建议在这些小鼠体内利用 DAPT 来改善纤毛功能，最终目标是改善中冶集团。我们的首要目标是通过增强 COPD 的新病理生理学途径线粒体功能和纤毛发生以改善 MCC。