Pre-Clinical Evaluation of IRE1beta as a Novel Therapeutic Target for Cystic Fibrosis Airway Mucus Production

IRE1beta 作为囊性纤维化气道粘液产生的新治疗靶点的临床前评估

基本信息

批准号：
10296058
负责人：
Carla Maria Pedrosa Ribeiro
金额：
$ 46.46万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-15 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10296058
关键词：
Address Affect Airway Disease Amino Acids Anti-Inflammatory Agents Asthma Attenuated Bacterial Infections Binding Binding Proteins Birth Bronchiectasis Bronchiolitis Cells Child Chronic Clinic Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Cytoplasmic Tail Data Distal Endoplasmic Reticulum Enzymes Epithelial Exhibits Feedback Gastrointestinal tract structure Generations Genetic Human Impairment Inflammation Inositol Integral Membrane Protein Interleukin-1 Lead MAPK8 gene MUC5AC gene MUC5B gene Mammals Mediating Medical Messenger RNA Modeling Mucins Mucous body substance Mus Obstruction Para-Influenza Virus Type 1 Pathway interactions Pharmacology Phosphotransferases Preclinical Testing Production Protein Isoforms Proteins Pulmonary Cystic Fibrosis Pulmonary Fibrosis RNA Splicing Regulation Respiratory Syncytial Virus Infections Respiratory System Respiratory syncytial virus Ribonucleases Role Signal Pathway Structure TNF gene Tertiary Protein Structure Testing Therapeutic Effect VX-770 Viral Bronchiolitis Virulence Factors Virus Diseases airway epithelium airway inflammation bronchial epithelium cystic fibrosis airway cystic fibrosis airway epithelia cystic fibrosis mouse cystic fibrosis patients cytokine efficacy testing endoplasmic reticulum stress idiopathic pulmonary fibrosis infant infection inhibitor/antagonist innovation mutant neutrophil new therapeutic target novel therapeutics overexpression p38 Mitogen Activated Protein Kinase preclinical evaluation prevent respiratory infection virus response sensor targeted treatment transcription factor

项目摘要

Cystic fibrosis (CF) lungs exhibit mucoinflammatory responses soon after birth, likely triggered by viral infections and/or aspiration. Respiratory syncytial virus (RSV) causes bronchiolitis leading to airway muco- obstruction in young CF children, who exhibit increases in MUC5B and MUC5AC mucins in their airways. Because no treatments are available for CF airway mucus overproduction, there is a clear unmet medical need for therapies that target mucin synthesis in CF airways. CF airway epithelial inflammation triggers endoplasmic reticulum (ER) stress and activates the inositol requiring enzyme 1 (IRE1), which exists in two isoforms, α and β. IRE1α is ubiquitous, but IRE1β is only expressed in mucous cells of the respiratory and GI tracts. We have shown that IRE1β (but not IRE1α) is required for airway mucin production. IRE1 is an ER transmembrane protein with a lumenal domain (sensor of unfolded proteins) and a cytoplasmic domain (effector) with kinase and RNase activities. It is unknown whether the IRE1 lumenal domain senses unfolded mucins and whether its cytoplasmic domains mediate mucin production; however, our previous studies suggested that mucin production triggers ER stress and activates IRE1β kinase-induced RNase activation. Because the activated IRE1 RNase splices the mRNA of X-box binding protein-1 (XBP-1s), a transcription factor that up-regulates mucin production, this may provide a mechanism for CF airway epithelial mucin overproduction. In non-mucous cells, IRE1 kinase activates JNK, p38 MAP kinase and NF-B via protein interactions, but it is unknown whether the IRE1 kinase activates these pathways, which are relevant to CF airways because they can promote mucin production. Our preliminary data indicate that IRE1β, MUC5AC and MUC5B levels are up-regulated in native CF human airways and in freshly isolated CF human distal airway epithelia. Over-expression of wild type IRE1 in primary human bronchial epithelia (HBE) increased mucin production, whereas over-expression of IRE1 mutants that lack kinase and/or RNase activities decreased mucin production. IL-1 and TNF, predominant CF airway cytokines, differently affected XBP-1s (only IL-1 increased XBP-1s) and mucin production (IL-1 > TNF) in HBE, suggesting that they up-regulate mucin production via IRE1β RNase-dependent and independent mechanisms. KIRA6, an IRE1 kinase + RNase inhibitor, blunted IL-1-increased XBP-1s and mucins in CF HBE. Notably, a combination of CFTR modulators (VX-445, VX-661 and VX-770) had no effect on mucin production. RSV infection of HBE increased XBP-1s and mucin production/secretion, and these responses were blunted by KIRA6. Finally, murine parainfluenza virus type 1-infected mice developed viral bronchiolitis and airway muco-obstruction, modeling the bronchiolitis in CF infants infected by RSV. Our aims will test the role of IRE1β protein domains in CF airway cytokine-increased mucin production in CF HBE. We will also evaluate the therapeutic effect of IRE1β inhibition on respiratory virus infection-induced mucin production in CF HBE and murine airways, including a CF mouse. Our studies may lead to the generation of IRE1β inhibitors as novel therapeutics for CF airway muco-obstruction.

囊性纤维化（CF）肺在出生后不久暴露于粘液炎症反应，可能是由病毒引起的感染和/或抽吸。呼吸道合胞病毒（RSV）引起的细支气管炎，导致气道粘液年轻的CF儿童的阻塞，他们的呼吸道中的MUC5B和MUC5AC粘蛋白暴露了增加。因为没有治疗可用于CF气道粘液过量生产，因此有明显的未满足的医疗需求用于针对CF气道中粘蛋白合成的疗法。 CF气道上皮注入触发内质网状（ER）应力并激活需要酶1（IRE1）的肌醇，该酶存在于两个同工型中，α和 β。 IRE1α无处不在，但IRE1β仅在呼吸道和胃肠道的粘液细胞中表达。我们有表明气道粘蛋白产生需要IRE1β（但不是IRE1α）。 IRE1是ER跨膜蛋白带有腔域（展开蛋白的传感器）和具有激酶和RNase的细胞质结构域（效应子）活动。尚不清楚IRE1腔内结构域是否会感觉到展开的粘蛋白以及其细胞质是否域介导粘蛋白的产生；但是，我们以前的研究表明粘蛋白产生触发ER 应力并激活IRE1β激酶诱导的RNase激活。因为激活的IRE1RNase拼接 X-box结合蛋白-1（XBP-1）的mRNA，这是一种上调粘蛋白产生的转录因子，今年5月提供CF气道上皮粘蛋白过量生产的机制。在非糖细胞中，IRE1激酶激活 JNK，p38映射激酶和NF-B通过蛋白质相互作用，但是尚不清楚IRE1激酶是否激活这些途径与CF气道相关，因为它们可以促进粘蛋白产生。我们的初步数据表明IRE1β，MUC5AC和MUC5B水平在天然CF人类气道中上调新鲜分离的CF人类远端气道上皮。原发性支气管中野生型IRE1的过表达上皮（HBE）增加了突变的产生，而缺乏激酶和/或的IRE1突变体的过表达 RNase活性降低了粘蛋白的产生。 IL-1和TNF，主要的CF气道细胞因子，以不同的方式在HBE中影响XBP-1（仅IL-1增加XBP-1S）和粘蛋白的产生（IL-1>TNF），这表明他们通过IRE1βRNase依赖性和独立机制上调粘蛋白产生。 Kira6，一个IRE1 激酶 + RNase抑制剂，CF HBE中的IL-1XBP-1和粘蛋白钝化。值得注意的是， CFTR调节剂（VX-445，VX-661和VX-770）对粘蛋白的产生没有影响。 HBE的RSV感染 XBP-1S和粘蛋白产生/分泌增加，这些反应被KIRA6钝化。最后，鼠副1.一种感染的小鼠帕林弗鲁内扎病毒开发了病毒细支气管炎和气道粘液粘结膜，建模 RSV感染的CF婴儿的支气管炎。我们的目标将测试IRE1β蛋白质结构域在CF气道中的作用 CF HBE中的细胞因子吸收的粘蛋白产生。我们还将评估IRE1β抑制的治疗作用在呼吸道病毒感染引起的CF HBE和鼠气中的粘蛋白产生（包括CF小鼠）。我们的研究可能导致IRE1β抑制剂的产生，作为CF气道粘液粘结的新疗法。