Endoplasmic Reticulum Stress Signaling in Allergen-induced Airway Remodeling

过敏原诱导的气道重塑中的内质网应激信号传导

• 批准号: 10394300
• 负责人: Vikas Anathy
• 金额: $ 39万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10394300
• 关键词: ATF6 gene; Ablation; Adenoviruses; Allergens; Allergic; Asthma; Attenuated; Binding; Biochemical; Biological Assay; CRISPR/Cas technology; Cell Culture Techniques; Cell Line; Cells; Complement; Cysteine; Data; Development; Disease; Dissociation; Disulfides; Endoplasmic Reticulum; Enzymes; Eosinophilia; Epithelial; Epithelial Cells; Extrinsic asthma; Fibrosis; GRP78 gene; Genes; Genetic Transcription; Human; Inflammation; Inflammatory; Inflammatory Response; Knockout Mice; Light; Lung; Mediating; Mediator of activation protein; Modality; Modification; Molecular Chaperones; Mus; Organelles; Oxidation-Reduction; Oxygen; Pathogenesis; Pathway interactions; Pharmacology; Process; Production; Protein Disulfide Isomerase; Proteins; Pulmonary Pathology; Reactive Oxygen Species; Reporting; Resistance; Resolution; Role; Signal Transduction; Stress; Suggestion; Supportive care; System; Time; Transducers; Transgenic Mice; Translations; Up-Regulation; airway epithelium; airway hyperresponsiveness; airway inflammation; airway remodeling; allergic airway disease; allergic response; alternative treatment; asthma model; asthmatic; base; chemokine; conventional therapy; cytokine; dimer; disulfide bond; efficacy evaluation; endoplasmic reticulum stress; inhibitor; insight; lung development; monomer; mortality; mouse model; mutant; novel; oxidation; pulmonary function; response; small hairpin RNA; therapeutic target; treatment strategy

PROJECT SUMMARY Severe allergic asthma is marked by airway inflammation, remodeling, and hyperresponsiveness, which correlates with decreased lung function and increased mortality. We have identified the endoplasmic reticulum (ER) based unfolded protein response (UPR) as a critical mediator of lung epithelial inflammatory and fibrotic responses to allergens. Recent reports highlight that increases in the UPR pathways are potentially classified as an endotype of severe asthma. However, the mechanisms whereby the UPR response is initiated and perpetuated in settings of severe asthma remain unaddressed, and will be the focus of this application. Our novel preliminary results now suggest that allergen challenge increases reactive oxygen species (ROS) in the ER and oxidative modification of cysteines in GRP78. Oxidation of GRP78 resultes in dissociation of GRP78 and causes initial activation of the UPR transducer, ATF6. We observed that the ATF6 target, PDIA5, was also significantly upregulated in both asthmatics and a mouse asthma model in association with an enhanced allergic responses in lungs. Intriguingly, PDIA5 reduces cysteines disulfides to (-S-S-) to sulfhydryls (-SH) of ATF6, which is suggestive of a feed-forward regulatory mechanism to sustain the UPR. Based on these observations, we hypothesize that allergen-induced oxidation of GRP78 initiates the UPR signaling, with subsequent increases in PDIA5 prolong the UPR and thereby increasing allergic airway responses. To examine this hypothesis, we propose the following specific aims: In Specific Aim #1 we will determine the functional role of allergen-induced reactive oxygen species and subsequent oxidation of GRP78 in initiation of the UPR, expression of cytokines/chemokines, and subsequent induction of pro-inflammatory response and development of lung remodeling. The specific Aim #2 seeks to dissect the critical requirement of allergen-induced PDIA5 in disulfide mediated processing of a transducer of UPR ATF6 in severe allergic airway responses. In both aims we will use transgenic mouse models, cell culture and sensitive redox and biochemical assays. Most importantly we will examine the efficacy of specific inhibitors of ATF6α (Ceapin-A7) and PDIA5 (LOC14) in attenuating allergen-induced UPR, decreasing subsequent pro- inflammatory responses, and ultimately resulting in resolution of allergen-induced lung pathology. These studies will shed light on the importance of the allergen-induced lung epithelial UPR in pro-inflammatory responses, lung remodeling and offers insight into new and highly needed treatment modalities for severe allergic airway disease beyond supportive care.

项目摘要 严重的过敏性哮喘以气道注射，重塑和反应性高度标记，这 与肺功能降低和死亡率增加相关。我们已经确定了内质网 基于（ER）的展开蛋白质反应（UPR）是肺上皮炎症和纤维化的关键介质 对过敏原的反应。最近的报告强调，UPR途径的增加可能被归类为 严重哮喘的内型。但是，启动UPR响应的机制以及 在严重哮喘的环境中永久存在，这将是该应用程序的重点。 我们的新型初步结果现在表明，过敏原挑战增加了活性氧（ROS） GRP78中半胱氨酸的ER和氧化修饰。 GRP78的氧化导致GRP78的解离 并导致UPR传感器ATF6的初始激活。我们观察到ATF6目标PDIA5是 在哮喘和小鼠哮喘模型中也显着上调 肺中的过敏反应。有趣的是，PDIA5将半胱氨酸二硫化物降低至（-s-s-）至硫酸（-sh）的（-sh） ATF6提出了一种维持UPR的喂养法规机制。基于这些 观察结果，我们假设过敏原诱导的GRP78氧化会启动UPR信号，并随着UPR信号传导 随后的PDIA5增加延长了UPR，从而增加了过敏性气道响应。检查 这个假设，我们提出了以下具体目的： 在特定目标1中，我们将确定过敏原诱导的活性氧和 GRP78随后在UPR启动时的氧化，细胞因子/趋化因子的表达以及随后的 诱导促炎反应和肺重塑的发展。具体目标＃2试图 在二硫键介导的处理器处理中，剖析过敏原诱导的PDIA5的关键要求 在严重的过敏性气道反应中UPRATF6。在这两个目标中，我们都将使用转基因小鼠模型，细胞培养 以及敏感的氧化还原和生化测定。最重要的是，我们将检查特定抑制剂的效率 在衰减过敏原诱导的UPR时，ATF6α（Ceapin-A7）和PDIA5（LOC14）的of ofaTF6α（loc14） 炎症反应，并最终导致过敏原诱导的肺病理的解决。这些研究 会阐明过敏原诱导的肺上皮UPR在促炎反应中的重要性，肺 重塑并深入了解严重过敏性气道疾病的新的且急需的治疗方式 超越支持性护理。