Chloride Channel Regulators, Calcium Activated (CLCA) Proteins in Equine Asthma - Mechanistic Relevance as Immunomodulators and Potential Mucus Biomarkers?

马哮喘中的氯离子通道调节剂、钙激活 (CLCA) 蛋白 - 作为免疫调节剂和潜在粘液生物标志物的机制相关性？

• 批准号: 508761433
• 负责人: Privatdozent Dr. Lars Mundhenk
• 金额: --
• 依托单位: Institut für Tierpathologie (WE12)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/508761433?language=en
• 关键词: Chloride; Channel; Regulators; Calcium; Activated

Equine asthma is among the most important pulmonary diseases of horses with many current challenges regarding therapy and diagnostics. In addition to the incomplete knowledge on its development and sustainable therapeutic options of this distressing disease, there is still a lack of suitable biomarkers for target-oriented diagnostics. The project proposed here aims to link research resources in a networked, interdisciplinary approach with a cross-regional strategy to provide impetus for improved treatment of this common disease. Our preliminary work and the contributions of other research groups suggest that members of the chloride channel regulator, calcium-activated (CLCA) protein family may play a central role in the transmigration of neutrophil granulocytes into the airways in humans with asthma and corresponding mouse models via stimulation of alveolar macrophages. Furthermore, some representatives are overexpressed in airway diseases with mucus overproduction. Except for one representative, CLCA1, which is secreted by airway mucin-producing cells, the CLCA family is unknown in the horse to date. Translation of findings from other species is impeded by several inter-species differences known for this family. The goals of this project include (1) the characterization of the CLCA proteins in the horse and their occurrence in the airways, (2) deciphering their role as signaling molecules for neutrophil influx, (3) the identification of CLCA1 interaction partners and regulatory networks in the mucus that play a role in immune cell influx, and (4) the exploration of their potential as biomarkers in the mucus. First, we aim to identify CLCA members expressed in the airways of healthy and asthmatic horses. Equine alveolar macrophages will be stimulated with recombinant CLCA proteins or their functional domains to determine their response using global RNA and protein technologies and to uncover protein structure-activity relationships. Furthermore, we will test whether the immunostimulatory response of macrophages results in neutrophil influx in vitro. For the remaining objectives, proteomics technologies will be used to comparatively quantify the total proteome specifically in the mucus of bronchoalveolar lavage fluid between healthy horses and those with equine asthma. Here, CLCA proteins and other proteins will be differentially quantified in the landscaping approach in the context of the clinical picture to assess their biomarker potential and discover protein-protein interaction networks. The results will help to (1) better understand the development of neutrophilic bronchiolitis in equine asthma, (2) identify suitable mucus proteins as biomarkers for improved diagnostics, and (3) potentially explore new therapeutic approaches.

马哮喘是马最重要的肺部疾病之一，目前在治疗和诊断方面面临许多挑战。除了对其发展和这种令人痛苦的疾病的可持续治疗选择的不完全了解之外，仍然缺乏适合靶向诊断的生物标志物。这里提出的项目旨在将网络化、跨学科方法中的研究资源与跨区域战略联系起来，为改进这种常见疾病的治疗提供动力。我们的初步工作和其他研究小组的贡献表明，氯离子通道调节剂钙活化（CLCA）蛋白家族成员可能在中性粒细胞通过刺激肺泡巨噬细胞进入哮喘患者和相应小鼠模型的气道中发挥核心作用。此外，一些代表在粘液分泌过多的气道疾病中过度表达。除了一种具有代表性的CLCA1，它是由气道粘液生成细胞分泌的，CLCA家族迄今在马中是未知的。其他物种的研究结果的翻译受到该科已知的几种物种间差异的阻碍。本项目的目标包括：(1)马的CLCA蛋白的特征及其在气道中的发生，(2)解释它们作为中性粒细胞内流的信号分子的作用，(3)鉴定在免疫细胞内流中起作用的黏液中的CLCA1相互作用伙伴和调节网络，以及(4)探索它们作为黏液生物标志物的潜力。首先，我们的目标是确定在健康和哮喘马的气道中表达的CLCA成员。将用重组CLCA蛋白或其功能域刺激马肺泡巨噬细胞，利用全局RNA和蛋白质技术确定它们的反应，并揭示蛋白质的结构-活性关系。此外，我们将在体外测试巨噬细胞的免疫刺激反应是否会导致中性粒细胞内流。对于剩下的目标，蛋白质组学技术将用于比较量化健康马和马哮喘患者之间支气管肺泡灌洗液粘液中的总蛋白质组。在这里，CLCA蛋白和其他蛋白将在临床图像的背景下在景观方法中进行差异量化，以评估其生物标志物潜力并发现蛋白质-蛋白质相互作用网络。该结果将有助于(1)更好地了解马哮喘中性粒细胞细支气管炎的发展，(2)确定合适的粘液蛋白作为生物标志物，以改进诊断，以及(3)潜在地探索新的治疗方法。