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）的特征是气流阻塞，粘液增加生产和易于复发的下呼吸道感染。它影响了约5％的美国人口排名第三，是死亡率的原因。这种高患病率和疾病的慢性导致经常住院，需要终身治疗。尽管有这种高疾病负担，但在过去的二十年中，没有新的FDA批准疗法。慢性香烟烟暴露，最常见的 COPD的原因，导致活性氧（ROS）的产生增加，线粒体降低能够处理这些ROS和细胞死亡的能力，后者导致肺泡表面崩溃，吹散，并减少肺表面以进行足够的氧气交换。纤毛是含微管蛋白的头发在配位的气管支气管树的上皮细胞的细胞表面投影从肺部扫除吸入污染物和病原体的基因波。在COPD中，这些纤毛是稀疏，发育迟缓和频率低于健康肺部的较低（MCC）和经常感染。由于压倒性ROS产生引起的线粒体功能障碍也是与缺陷的纤毛形成有关。我们先前的工作导致我们确定了12个氨基酸肽由于其能力在周围后转导正常的小鼠心脏组织，因此被称为心脏靶向肽注射。具有连续的单丙氨酸取代的丙氨酸扫描导致发现了两个丙氨酸突变体（S7a和R11a在位置7处具有丝氨酸，在第11位的精氨酸用丙氨酸代替）外周注射后心脏不强烈地转导肺上皮组织。主要参与者纤毛生成是Notch，这是一种转录因子，它决定了多能俱乐部细胞的命运和分化为产生杯状细胞的粘液为代价多丝硅细胞。我们已经证明了回收的处理小鼠气管和人类鼻皮细胞培养物，具有2nm Dapt（n- [n-（3,5-二氟苯乙基）-L-）小分子缺口抑制剂，丙基甘氨酸T-苯基甘氨酸T-丁基酯，导致一定程度的显着增加与对照组相比，纤毛生成，纤毛长度和纤毛搏动频率。在这笔赠款中，我们建议开发这些新颖的肺靶向肽作为向量，以提供许多不同的ROS清除剂（szeto-shiller肽，mitotempo，降低谷胱甘肽）到具有烟雾引起的COPD的小鼠的肺。我们是还建议在这些小鼠中利用体内DAPT，以改善睫状功能，以实现最终目标改善MCC。我们的总体目标是通过增强COPD中的新型病理生理途径线粒体功能和纤毛发生以改善MCC。