Restoring mucociliary clearance apparatus to mitigate lung inflammation in the context of HIV and cigarette smoke

恢复粘膜纤毛清除装置以减轻艾滋病毒和香烟烟雾背景下的肺部炎症

• 批准号: 10664021
• 负责人: Srinivasan Chinnapaiyan
• 金额: $ 14.75万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10664021
• 关键词: 3' Untranslated Regions; Abbreviations; Accounting; Affect; Air; Airway Disease; Allergens; Attenuated; Bacteria; Bacterial Infections; Bacterial Pneumonia; Bronchoalveolar Lavage; Cells; Chronic; Chronic Bronchitis; Chronic Obstructive Pulmonary Disease; Cilia; Clustered Regularly Interspaced Short Palindromic Repeats; Compensation; Core-Binding Factor; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Deacetylase; Development; Disease; Epithelial Cells; Etiology; Event; Feedback; Frequencies; Functional disorder; Gene Expression; Genes; Goals; Goblet Cells; HIV; HIV Receptors; Human; Hyperplasia; Immediate-Early Proteins; Impairment; Incidence; Individual; Inflammation; Inflammatory; Intervention; Lead; Liquid substance; Lung; Lung diseases; Lung infections; MUC5AC gene; Mediating; MicroRNAs; Morbidity - disease rate; Mucociliary Clearance; Mucous body substance; Pathologic; Pathology; Pathway interactions; Patients; Peptide Hydrolases; Persons; Play; Publishing; Pulmonary Inflammation; Quality of life; Recurrence; Respiratory Tract Infections; Risk; Risk Factors; Role; SIRT1 gene; Severities; Signal Transduction; Site; Smoker; Smoking; Smoking Status; System; TGFB1 gene; Testing; Thinness; Untranslated Regions; Up-Regulation; Viral Load result; airway epithelium; airway surface liquid; antagonist; antiretroviral therapy; bronchial epithelium; chronic respiratory disease; cigarette smoke; cilium biogenesis; comorbidity; cytokine; exposure to cigarette smoke; improved; lung microbiome; microbial colonization; mortality; mucus clearance; mucus hypersecretion; notch protein; novel; pathogen; pollutant; preservation; pulmonary function; pulmonary function decline; respiratory colonization; therapeutic evaluation

PROJECT SUMMARY People living with HIV demonstrate increased incidence of lung inflammation and HIV is an independent risk factor development of COPD. HIV Tat, an immediate early protein of HIV and cigarette smoke suppress the deacetylase SIRT1 that regulates ADAM17, a protease involved in activating Notch signaling as well as proteolytic cleavage and activation of proinflammatory cytokines. This can lead to downstream deleterious effects on ciliogenesis, mucociliary clearance and lung function decline. While chronic inflammation in COPD is a multifactorial etiology, impaired mucociliary clearance leading to recurrent lung infections can play an important role in promoting lung inflammation. Optimal mucociliary clearance requires mucus, cilia, and a thin layer of airway surface liquid to facilitate ciliary beating. Abnormalities in any compartment of the mucociliary system can compromise mucus clearance leading to mucus impaction entrapping bacteria and promoting chronic bacterial infection. HIV Tat promotes an aberrant microRNAome in the primary bronchial epithelium and upregulates miR-142-5p, a microRNA known to suppress SIRT1. This proposal will identify mechanisms involved in HIV Tat promotes mucociliary dysfunction and identifying individual effects on ciliogenesis, mucus hypersecretion and inflammation. Given the causal role for SIRT1 suppression in mediating these effects, we will use a novel CRISPR-mediated gene specific microRNA antagonism approach to disrupt the miR-142-5p target site on the SIRT1 3’UTR. This will preserve SIRT1 expression in the context of HIV Tat and cigarette smoke without affecting other functions of miR-142-5p. Aim 1: HIV Tat upregulates miR-142-5p and suppress SIRT1 exacerbate upregulates ADAM17 levels by HIV and cigarette smoke exposure in NHBE ALI cultures. Given that HIV Tat and cigarette smoke mediated miR-142-5p dysregulation leading to SIRT1 suppression and aberrant Notch signaling. Specifically, we will analyze ADAM17 upregulation, ciliogenesis, goblet cell hyperplasia and ciliary beat frequency in NHBE ALI cultures. Aim 2: Since HIV Tat and cigarette smoke suppresses SIRT1, and SIRT1 suppression impairs mucociliary clearance apparatus, we will determine if gene-specific microRNA antagonism in combination with SIRT1 activator to rescue SIRT1 levels, suppression of inflammation and enhance mucociliary clearance apparatus in primary bronchial epithelial cells treated with HIV Tat and exposed to cigarette smoke.

项目摘要 HIV感染者肺部炎症的发病率增加，HIV是一种独立的风险 COPD的发展因素。HIV达特是HIV的一种早期蛋白质，香烟烟雾抑制了HIV的免疫应答。 调节ADAM 17的脱乙酰酶SIRT 1，ADAM 17是一种参与激活Notch信号传导的蛋白酶， 蛋白水解裂解和促炎细胞因子的活化。这可能导致下游有害的 对纤毛生成、粘膜纤毛清除和肺功能下降的影响。虽然COPD中的慢性炎症是 一种多因素病因，导致复发性肺部感染的粘膜纤毛清除受损可能在 促进肺部炎症。最佳的粘膜纤毛清除需要粘液、纤毛和一层薄薄的 气道表面液体，以促进纤毛跳动。在粘膜纤毛系统的任何隔室中的脱落可以 损害粘液清除率，导致粘液嵌塞，从而截留细菌并促进慢性细菌感染， 感染 HIV达特促进原代支气管上皮中的异常microRNAome并上调miR-142- 5 p， 一种抑制SIRT 1的微小RNA这项建议将确定参与艾滋病毒达特促进机制 粘液纤毛功能障碍，并确定对纤毛发生、粘液分泌过多和 炎症考虑到SIRT 1抑制在介导这些效应中的因果作用，我们将使用一种新的 CRISPR介导的基因特异性microRNA拮抗方法破坏miR-142- 5 p靶位点， SIRT1 3'UTR.这将在HIV达特和香烟烟雾的背景下保留SIRT 1表达，而不影响SIRT 1的表达。 miR-142- 5 p的其他功能目的1：HIV达特上调miR-142- 5 p并抑制SIRT 1的恶化 在NHBE ALI培养物中通过HIV和香烟烟雾暴露上调ADAM 17水平。鉴于艾滋病毒达特和 香烟烟雾介导的miR-142- 5 p失调导致SIRT 1抑制和异常Notch信号传导。 具体而言，我们将分析ADAM 17上调，纤毛发生，杯状细胞增生和纤毛搏动， NHBE ALI培养物中的频率。目的2：由于HIV达特和香烟烟雾抑制SIRT 1， 抑制损害粘膜纤毛清除装置，我们将确定是否基因特异性microRNA拮抗 与SIRT 1激活剂组合以拯救SIRT 1水平，抑制炎症并增强 用HIV达特处理并暴露于 香烟烟雾。