Role of Mucin in Lung Homeostasis and Pathophysiology

粘蛋白在肺稳态和病理生理学中的作用

• 批准号: 10115780
• 负责人: Christopher M Evans
• 金额: $ 59.65万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10115780
• 关键词: Acute; Affect; Air Movements; Allergic; Anabolism; Animals; Applied Genetics; Area; Asthma; Automobile Driving; Award; Biology; Biomedical Engineering; Breathing; C-terminal; CRISPR/Cas technology; Cell Culture Techniques; Cells; Complement; Cysteine; Data; Development; Devices; Disease; Disulfides; Economics; Exposure to; Extrinsic asthma; Face; Functional disorder; Gel; Glycoproteins; Health; Health Care Costs; Homeostasis; Human; Impairment; Inhalation; Interleukin-13; Intervention; Knockout Mice; Laboratories; Link; Lung; Lung diseases; MUC5AC gene; MUC5B gene; Mediating; Modeling; Molecular; Mucins; Mucociliary Clearance; Mucolytics; Mucous body substance; Mus; Mutant Strains Mice; Mutation; N-terminal; Obstruction; Pathologic; Personal Satisfaction; Phenotype; Physiological; Point Mutation; Polymers; Positioning Attribute; Prevention; Prevention strategy; Property; Protein Isoforms; Proteins; Reducing Agents; Research; Rheology; Role; Sampling; Severity of illness; Sputum; Technology; Testing; Therapeutic Agents; Therapeutic Uses; Thick; Thinness; Transgenic Mice; airway epithelium; airway hyperresponsiveness; cytokine; disulfide bond; effective therapy; extracellular; improved; in vivo; innovation; macromolecular assembly; macromolecule; novel; novel strategies; particle; polymerization; preservation; prevent; public health relevance; tool; viscoelasticity; von Willebrand Factor

PROJECT SUMMARY/ABSTRACT Although mucus is a defensive barrier that is critical for maintaining healthy lungs, excessive mucus is a significant pathological contributor to lung diseases. The disparate faces of mucus function and dysfunction epitomize the need for tightly controlling the predominant macromolecules in airway mucus - the polymeric mucins MUC5AC and MUC5B. In asthma, mucus viscoelasticity increases and causes airflow and mucociliary clearance (MCC) impairments that are not adequately treated. We postulate that by determining specific molecular mechanisms of MUC5AC and MUC5B biosynthesis, we will identify novel mechanisms and potential treatments for improving airway mucus function and dysfunction. We previously demonstrated significant functions for Muc5ac and Muc5b in knockout mice. We now seek to interrogate how Muc5ac and Muc5b function as polymers. Muc5ac and Muc5b assemble by inter-molecular disulfide bonding at their cysteine-rich carboxyl (C-) and amino (N-) termini, resulting in the formation of massive glycopolymers. Recently, we treated allergically inflamed mice with inhaled agents that disrupted mucin polymer disulfide bonds, and we found that mucus viscoelasticity, MCC, and airflow function all improved. While inhaled mucolytics could be useful, there are currently no safe or effective options available for therapeutic use. Thus, there is further need to determine MUC5AC and MUC5B isoform specific polymer functions. We hypothesize that mucus function and dysfunction are dictated by specific mucin isoform expression and disulfide polymerization mechanisms that regulate mucus viscoelastic gel and transport properties. We will test the following hypotheses in thee Specific Aims: 1) that isoform specific mucin polymers mediate AHR and MCC; 2) that polymerization is mediated by disulfide bonds between conserved cysteine residues in the N- and C-termini of MUC5AC/Muc5ac and MUC5B/Muc5b; and 3) that conserved N- and C-terminal cysteines are required for mucin isoform-specific polymerization and function in vivo. We will conduct studies in mucin mutant mice, in human mucus samples, in airway epithelial cell cultures, and with bioengineered Human Small Airway-on-a-Chip devices. Successful completion of the Aims proposed here will further advance our understanding of mucin polymer formation and function in the airways. Our studies could also identify improved strategies for preventing obstruction while preserving defensive mucus functions that are required for lung health.

项目概要/摘要 虽然粘液是一种防御屏障，对于维持肺部健康至关重要，但过多的粘液是一个重要的问题。 肺部疾病的病理因素。粘液功能和功能障碍的不同方面体现了 需要严格控制气道粘液中的主要大分子 - 聚合粘蛋白 MUC5AC 和 MUC5B。在哮喘中，粘液粘弹性增加并导致气流和粘液纤毛清除 (MCC) 未得到充分治疗的损伤。我们假设通过确定特定的分子机制 MUC5AC 和 MUC5B 生物合成，我们将确定新的机制和潜在的治疗方法来改善 气道粘液功能和功能障碍。我们之前演示了 Muc5ac 和 Muc5b 的重要功能 在基因敲除小鼠中。我们现在试图探究 Muc5ac 和 Muc5b 作为聚合物如何发挥作用。 Muc5ac 和 Muc5b 通过分子间二硫键在其富含半胱氨酸的羧基 (C-) 和氨基 (N-) 末端组装，从而产生 形成块状糖聚合物。最近，我们用吸入剂治疗过敏性发炎的小鼠， 破坏粘蛋白聚合物二硫键，我们发现粘液粘弹性、MCC 和气流都起作用 改善了。虽然吸入粘液溶解剂可能有用，但目前没有安全或有效的选择 治疗用途。因此，还需要确定MUC5AC和MUC5B亚型特定的聚合物功能。 我们假设粘液功能和功能障碍是由特定的粘蛋白亚型表达和 调节粘液粘弹性凝胶和运输特性的二硫化物聚合机制。我们将测试 具体目标中的以下假设： 1) 同种型特异性粘蛋白聚合物介导 AHR 和 MCC； 2）那个 聚合是由 N 端和 C 端保守的半胱氨酸残基之间的二硫键介导的 MUC5AC/Muc5ac 和 MUC5B/Muc5b； 3) 粘蛋白需要保守的 N 端和 C 端半胱氨酸 同种型特异性聚合和体内功能。我们将在粘蛋白突变小鼠、人类中进行研究 粘液样本、气道上皮细胞培养物以及生物工程人类小气道芯片设备。 成功完成此处提出的目标将进一步增进我们对粘蛋白聚合物的理解 气道的形成和功能。我们的研究还可以确定预防阻塞的改进策略 同时保留肺部健康所需的防御性粘液功能。