The role of PUMA in the progression of cigarette smoking-induced COPD

PUMA 在吸烟引起的 COPD 进展中的作用

• 批准号: 10930186
• 负责人: Dongshi Chen
• 金额: $ 41.4万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-22 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10930186
• 关键词: Air; Alveolar; Animal Model; Apoptosis; Apoptosis Inhibitor; Apoptotic; Attenuated; BCL2 gene; Bronchoalveolar Lavage; Cause of Death; Cell Death; Cell Death Induction; Cell Death Inhibition; Cells; Chronic Obstructive Pulmonary Disease; Chronic lung disease; Cigarette smoke-induced emphysema; Data; Databases; Development; Disease Progression; Elasticity; Epithelial Cells; Etiology; Evaluation; Exposure to; Family member; Foundations; Genomics; In Vitro; Induction of Apoptosis; Inflammation; Inflammatory; Inflammatory Response; Inhibition of Apoptosis; Investigation; Knockout Mice; Liquid substance; Lung; Lung diseases; Macrophage; Macrophage Activation; Mediating; Mitochondria; Modeling; Molecular; Molecular Weight; Mus; Organoids; Pathogenesis; Patients; Pattern; Play; Pre-Clinical Model; Process; Pulmonary Emphysema; Pulmonary Inflammation; Reporting; Research; Role; Signal Pathway; Smoking; Structure of parenchyma of lung; TNF gene; TP53 gene; Testing; Therapeutic Uses; Tissues; airway obstruction; cigarette smoke; cigarette smoke-induced; cigarette smoke-induced COPD; cigarette smoking; cytokine; design; effective therapy; experimental study; exposure to cigarette smoke; improved; in vivo; inhibitor; knock-down; loss of function; mortality; neutrophil; novel; novel therapeutic intervention; novel therapeutics; pharmacologic; pre-clinical; prevent; pulmonary function; small molecule inhibitor; smoking cessation; targeted treatment; therapeutic target

Abstract Chronic obstructive pulmonary disease (COPD) is the fourth-leading cause of mortality worldwide. The causes of COPD are usually determined by multiple factors, among which exposure to cigarette smoke (CS) is the most common cause of COPD. CS exposure has been reported to induce epithelial cell death by apoptosis and necroptosis, and trigger lung inflammation. Most previous studies on cell death focus on the mechanism of apoptosis. Necroptosis, a new form of cell death, has emerged as another important mechanism for COPD pathogenesis. Therefore, revealing a novel target for both apoptosis and necroptosis will help the development of new treatments for COPD. Our preliminary data show that p53 up-regulated modulator of apoptosis (PUMA), a pro-apoptotic BH3-only Bcl-2 family member, is markedly induced in the lung tissues of COPD patients and mice exposed to CS treatment, as well as in lung epithelial cells treated with air-liquid interface (ALI) CS exposure or cigarette smoking extract (CSE). PUMA deficiency protected against CSE-induced apoptosis and necroptosis in the lung epithelial cells, and mouse alveolar organoids. Depletion of PUMA in the lung epithelial cells also attenuated the macrophage inflammatory profile. Moreover, knockdown of PUMA has higher suppressive effect on pro-inflammatory cytokines induction by CSE exposure than inhibition of cell death, indicating that PUMA might have a new function in triggering inflammation besides cell death. PUMA knock-out mice showed decreased lung inflammation, apoptosis, and necroptosis after 2 months of CS exposure, and rescued lung function loss after 6 months of CS exposure. Based on these preliminary findings, we hypothesize that PUMA mediates CS-induced apoptosis and necroptosis, and contributes to lung inflammation in the pathogenesis of COPD. We further hypothesize that PUMA can be pharmacologically targeted as a novel therapy against COPD. Specifically, we will investigate the functional role of PUMA in CS-induced lung epithelial cell death, as well as inflammatory macrophage activation, and test whether PUMA deletion or inhibition by a first-in-class small molecule inhibitor can protect the lung from CS-induced emphysema. In the proposed studies, we will pursue these three specific aims: (1) Test the hypothesis that PUMA orchestrates CS-induced apoptosis and necroptosis. (2) Test the hypothesis that PUMA induction in lung epithelial cells triggers pro-inflammatory cytokines expression and causes cell death independent lung inflammation. (3) Test the hypothesis that inhibition of PUMA by small molecular weight inhibitors prevents the development of cigarette smoke-induced emphysema in mice. This project is expected to reveal a new function of PUMA in the pathogenesis of COPD, especially its novel function in the induction of inflammatory cytokines. The completion of the proposed studies will provide proof-of- principle evidence for targeting PUMA as a novel therapeutic approach for COPD treatment.

摘要 慢性阻塞性肺疾病(COPD)是全球第四大死亡原因。原因何在 COPD的发病通常是由多种因素决定的，其中暴露于香烟烟雾(CS)是最多的 慢性阻塞性肺病的常见病因。据报道，CS暴露可通过细胞凋亡和细胞凋亡诱导上皮细胞死亡 坏死性下垂，并引发肺部炎症。以往对细胞死亡的研究大多集中在细胞死亡的机制上。 细胞凋亡。坏死性下垂是一种新的细胞死亡形式，已成为COPD的另一种重要机制 发病机制。因此，揭示细胞凋亡和坏死性下垂的新靶点将有助于该研究的发展。 治疗慢性阻塞性肺疾病的新疗法。我们的初步数据显示，P53上调了凋亡调节因子(PUMA)， 在COPD患者和慢性阻塞性肺疾病患者的肺组织中，显著诱导了BH3-Only Bcl2家族成员的促凋亡作用 暴露于CS处理的小鼠，以及暴露于气液界面(ALI)CS暴露的肺上皮细胞 或香烟烟抽提物(CSE)。彪马缺乏对CSE诱导的细胞凋亡和坏死性下垂的保护作用 在肺上皮细胞和小鼠肺泡上皮细胞中。肺上皮细胞中PUMA的耗竭也 减轻巨噬细胞的炎症反应。此外，PUMA的敲除具有较高的抑制作用 对CSE诱导的促炎细胞因子的作用优于对细胞死亡的抑制作用，表明PUMA 除了细胞死亡外，可能还具有引发炎症的新功能。美洲狮基因敲除小鼠显示 减少CS暴露2个月后的肺部炎症、细胞凋亡和坏死性下垂，挽救肺 CS暴露6个月后功能丧失。根据这些初步发现，我们假设PUMA 介导CS诱导的细胞凋亡和坏死性下垂，并在肺炎发病机制中参与肺部炎症 慢性阻塞性肺疾病（慢阻肺）。我们进一步假设PUMA可以作为治疗COPD的一种新的药物靶点。 特别是，我们将研究PUMA在CS诱导的肺上皮细胞死亡中的功能作用，以及 炎性巨噬细胞激活，并测试PUMA是否缺失或抑制由一流的小分子 分子抑制物对CS诱导的肺气肿有保护作用。在建议的研究中，我们会继续 这三个具体目标：(1)检验PUMA协调CS诱导的细胞凋亡和 坏死性下垂。(2)验证肺上皮细胞中PUMA诱导触发促炎作用的假说 细胞因子的表达和导致细胞死亡的非依赖性肺部炎症。(3)检验抑制假设 小分子量抑制剂PUMA预防香烟烟雾诱导的肺气肿 在老鼠身上。 本项目有望揭示PUMA在COPD发病机制中的新功能，特别是其新的功能。 在诱导炎性细胞因子中的作用。建议的研究完成后，将会证明- 以PUMA为靶点作为COPD治疗新方法的主要证据。