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: 10407568
• 负责人: Maliha Zahid
• 金额: --
• 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 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10407568
• 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 Pulmonary Disease; Cilia; 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; 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，粘液增加， 生产，并倾向于反复下呼吸道感染。它影响了约5%的美国人 人口，排名第三的死亡原因。这种高患病率和疾病慢性化导致 频繁住院，需要终身治疗。尽管疾病负担很高， 在过去的二十年里没有新的FDA批准的疗法。慢性吸烟暴露，最常见的 慢性阻塞性肺病的病因，导致活性氧（ROS）的产生增加，线粒体减少， 处理这些ROS的能力和细胞死亡，后者导致肺泡表面的破坏，起泡， 减少了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 气管支气管树的上皮细胞表面的突起以协调的方式跳动， 异时性波将吸入的污染物和病原体从肺部清除。在COPD中，这些纤毛 稀疏、发育不良、搏动频率低于健康肺，导致粘膜纤毛清除不良 (MCC)和反复感染。线粒体功能障碍，由于压倒性的ROS生产也是 与纤毛形成缺陷有关。我们先前的工作鉴定了一种12个氨基酸的肽， 称为心脏靶向肽，因为它能够在外周血中刺激正常小鼠心脏组织。 注射一个连续的丙氨酸扫描，单丙氨酸取代导致发现两个丙氨酸 突变体（S7 A和R11 A，其中第7位丝氨酸和第11位精氨酸被丙氨酸取代）， 而不是外周注射后心脏稳健转导的肺上皮组织。的关键角色 纤毛发生是Notch，一种决定多能性Club细胞命运和分化为 以多纤毛细胞为代价产生粘液的杯状细胞。我们已经证明， 用2nM DAPT（N-[N-（3，5-二氟苯乙酰基）-L- 丙氨酰]-S-苯基甘氨酸叔丁酯），一种小分子Notch抑制剂，导致了 纤毛发生、纤毛长度和纤毛搏动频率。在这项拨款中，我们建议 开发这些新的肺靶向肽作为载体，以递送许多不同的ROS清除剂 （Szeto-Schiller肽，Mitotempo，还原型谷胱甘肽）对患有吸烟诱导的COPD的小鼠的肺的作用。我们 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 改进MCC。我们的总体目标是通过增强COPD的新病理生理学途径， 线粒体功能和纤毛发生以改善MCC。