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: 10547928
• 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/10547928
• 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; 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; 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.

项目概要 艾滋病毒感染者肺部炎症的发生率增加，艾滋病毒是一种独立的风险 COPD 的因素发展。 HIV Tat 是 HIV 的一种早期蛋白质，吸烟可抑制 调节 ADAM17 的脱乙酰酶 SIRT1，ADAM17 是一种参与激活 Notch 信号传导的蛋白酶 促炎细胞因子的蛋白水解和激活。这可能会导致下游有害 对纤毛生成、粘液纤毛清除和肺功能下降的影响。虽然 COPD 的慢性炎症是 病因是多因素的，粘液纤毛清除受损导致反复肺部感染可能发挥重要作用 促进肺部炎症的作用。最佳的粘液纤毛清除需要粘液、纤毛和一层薄薄的 气道表面液体有利于纤毛跳动。粘液纤毛系统任何部分的异常都可以 损害粘液清除，导致粘液嵌塞，捕获细菌并促进慢性细菌 感染。 HIV Tat 促进初级支气管上皮中异常的 microRNAome 并上调 miR-142-5p， 已知可抑制 SIRT1 的 microRNA。该提案将确定涉及 HIV Tat 促进的机制 粘液纤毛功能障碍并确定对纤毛发生、粘液分泌过多和粘液纤毛的个体影响 炎。鉴于 SIRT1 抑制在介导这些影响中的因果作用，我们将使用一种新颖的方法 CRISPR介导的基因特异性microRNA拮抗方法破坏miR-142-5p靶位点 SIRT1 3’UTR。这将在 HIV Tat 和香烟烟雾的背景下保留 SIRT1 表达，而不影响 miR-142-5p 的其他功能。目标 1：HIV Tat 上调 miR-142-5p 并抑制 SIRT1 加剧 在 NHBE ALI 培养物中，HIV 和香烟烟雾暴露会上调 ADAM17 水平。鉴于 HIV Tat 和 香烟烟雾介导的 miR-142-5p 失调导致 SIRT1 抑制和异常的 Notch 信号传导。 具体来说，我们将分析 ADAM17 上调、纤毛发生、杯状细胞增生和纤毛跳动 NHBE ALI 培养物中的频率。目标 2：由于 HIV Tat 和香烟烟雾抑制 SIRT1，并且 SIRT1 抑制会损害粘膜纤毛清除装置，我们将确定基因特异性 microRNA 拮抗作用是否 与 SIRT1 激活剂联合使用可挽救 SIRT1 水平，抑制炎症并增强 用 HIV Tat 处理并暴露于的原代支气管上皮细胞中的粘液纤毛清除装置 香烟烟雾。