Development of tools and knowledge to facilitate the investigation of chloride channel regulator CLCA2 in rare human diseasses

开发工具和知识以促进罕见人类疾病中氯离子通道调节剂 CLCA2 的研究

• 批准号: 10728179
• 负责人: THOMAS John BRETT
• 金额: $ 15.55万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10728179
• 关键词: Airway Disease; Anions; Antibodies; Asthma; Binding; Biochemical; Biological; Biological Process; Brain; Calcium; Cell Culture Techniques; Cells; Chemosensitization; Chloride Channels; Chlorides; Chronic Obstructive Pulmonary Disease; Crohn' s disease; Cystic Fibrosis; Data; Diarrhea; Digestive System Disorders; Disease; Drug Targeting; Electrophysiology (science); Evaluation; Family; Family member; Future; Gastrointestinal Diseases; Gastrointestinal tract structure; Genetic; Goals; Health; Human; Inflammatory; Investigation; Knowledge; Ligands; Link; Literature; Lung; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Metalloproteases; Molecular; Monoclonal Antibodies; Mucociliary Clearance; Mucous Membrane; Pattern; Pilot Projects; Play; Protein Family; Proteins; Pulmonary Cystic Fibrosis; Rare Diseases; Reagent; Regulation; Research; Role; Route; Sampling; Stains; Structure; Surface; System; Therapeutic; Tissues; Trachea; Ulcerative Colitis; cystic fibrosis airway; design; experimental study; human disease; insight; knowledge base; mRNA Expression; member; novel; novel therapeutics; targeted treatment; tissue culture; tool; tool development

ABSTRACT: The calcium-activated chloride channel regulator CLCA2 has been associated with and suggested as a drug target for inflammatory diseases of the airway and gastrointestinal tract including the rare disease cystic fibrosis (CF). However, a detailed understanding of its biological functions has lagged far behind its association with these diseases, largely due to complete lack of biochemical knowledge and quality antibody reagents. We recently made groundbreaking advances investigating fellow family members, CLCA1 and CLCA4, by showing that they are self-cleaving metalloproteases that potentiate the calcium activated chloride channels TMEM16A and TMEM16B, respectively. In our preliminary data we show that CLCA2 can also potentiate a calcium- activated chloride channel in human cells. Therefore, we are uniquely poised to generate significant advances in our understanding of CLCA2 function in health and disease. This pilot project will develop the knowledge base and tools required to facilitate this. In Aim 1, we will investigate the mechanism of regulation of CLCA2 activity and comprehensively identify the TMEM16 family member modulated by CLCA2. In Aim 2, we will develop a novel panel of monoclonal antibody reagents specific for the mature, cleaved forms of CLCA2 (N- CLCA2 and C-CLCA2) that can be used in several experimental formats, and carry out initial analysis of healthy and diseased CF airway tissues and cell samples with them. The long-term goal of this project (beyond this pilot R03) is to develop a comprehensive mechanistic understanding of CLCA2 function in health and disease, which could provide new therapeutic routes to treat CF and other muco-obstructive diseases, as we demonstrate in our preliminary data. Successful completion of this pilot will provide the knowledge and tools to investigate the functional roles of CLCA2 in CF, with the potential to develop targeted therapeutics in subsequent projects, as well as provide insight into CLCA2 links in asthma, COPD, infectious diarrhea, Crohn’s disease, ulcerative colitis, and some cancers.

抽象的： 钙激活氯离子通道调节剂 CLCA2 与相关并被建议作为 气道和胃肠道炎症性疾病的药物靶标，包括罕见的囊性疾病 纤维化（CF）。然而对其生物学功能的详细了解却远远落后于其关联性 这些疾病的发生，很大程度上是由于完全缺乏生化知识和优质抗体试剂。我们 最近在调查其他家庭成员 CLCA1 和 CLCA4 方面取得了突破性进展， 它们是自裂解金属蛋白酶，可增强钙激活氯离子通道 TMEM16A 和TMEM16B，分别。在我们的初步数据中，我们表明 CLCA2 还可以增强钙- 激活人体细胞中的氯离子通道。因此，我们有能力取得重大进展 帮助我们了解 CLCA2 在健康和疾病中的功能。该试点项目将发展知识 促进这一点所需的基础和工具。在目标1中，我们将研究CLCA2的调节机制 活性并全面鉴定受 CLCA2 调节的 TMEM16 家族成员。在目标 2 中，我们将 开发一组新的单克隆抗体试剂，专门针对成熟、切割形式的 CLCA2（N- CLCA2 和 C-CLCA2) 可用于多种实验格式，并进行初步分析 健康和患病的 CF 气道组织及其细胞样本。该项目的长期目标（超越 该试点项目 R03）旨在对 CLCA2 在健康和 疾病，这可以为治疗 CF 和其他粘膜阻塞性疾病提供新的治疗途径，因为我们 在我们的初步数据中证明。该试点项目的成功完成将为以下人员提供知识和工具： 研究 CLCA2 在 CF 中的功能作用，有可能开发针对 CF 的靶向治疗方法 后续项目，以及深入了解 CLCA2 在哮喘、慢性阻塞性肺病、感染性腹泻、克罗恩病中的联系 疾病、溃疡性结肠炎和某些癌症。