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.

项目摘要 感染艾滋病毒的人表明肺部感染和艾滋病毒的发病率增加是一种独立的风险 COPD的因素发展。 HIV TAT，艾滋病毒和香烟烟的早期蛋白质抑制了 调节ADAM17的脱乙酰基酶SIRT1，这是一种激活Notch信号的蛋白酶以及 蛋白水解裂解和促炎细胞因子的激活。这可能导致下游有害 对纤毛生成，粘膜毛的清除和肺功能下降的影响。而COPD中的慢性炎症是 多因素的病因，受损的粘膜纤毛间隙导致肺部感染可以发挥重要作用 在促进肺部感染中的作用。最佳的粘膜纤毛清除需要粘液，纤毛和一层薄层 气道表面液体可促进纤毛跳动。在粘膜系统的任何隔间中，异常 损害损害清除率导致粘液撞击入口细菌并促进慢性细菌 感染。 HIV TAT促进了原发支气管上皮中的异常微洋小组，并上调miR-142-5p， 已知可以抑制SIRT1的microRNA。该建议将确定艾滋病毒TAT涉及的机制促进 粘毛功能障碍并确定对纤毛生成，粘液过度分泌和 炎。鉴于SIRT1抑制在介导这些效果中的因果作用，我们将使用一种新颖 CRISPR介导的基因特异性microRNA拮抗方法破坏了miR-142-5p靶点位点 Sirt1 3’utr。这将在HIV TAT和香烟烟的情况下保留SIRT1的表达 miR-142-5p的其他功能。目标1：HIV TAT上调miR-142-5p并抑制SIRT1恶化 NHBE Ali培养物中的艾滋病毒和香烟烟雾暴露于ADAM17水平。鉴于艾滋病毒tat和 香烟烟雾介导的miR-142-5p失调导致SIRT1抑制和异常凹口信号传导。 具体而言，我们将分析ADAM17上调，纤毛生成，杯状细胞增生和纤毛Beat NHBE Ali文化中的频率。 AIM 2：由于HIV TAT和香烟烟雾抑制SIRT1，而SIRT1 抑制会损害粘膜钙清除仪，我们将确定基因特异性microRNA拮抗作用是否存在 结合SIRT1激活剂来挽救SIRT1水平，抑制注射和增强 用HIV TAT处理并暴露于原发支气管上皮细胞中的粘膜纤毛清除设备 香烟烟。