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

• 批准号: 10026871
• 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
• 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/10026871
• 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; Epithelium; 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

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多种不同的 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。