Mucus Biochemistry and Biophysics Core

粘液清除异常是囊性纤维化病理表型的重要因素。 我们在过去十年的研究表明，CFTR与CF相关的功能障碍导致了脱水 呼吸道粘液层。这种脱水的结果是生物物理上的重大变化。 粘液层的性质及其与细胞表面的相互作用，即产生粘着的粘液 到上皮细胞，纤毛和咳嗽介导的粘液清除减慢/失败。要了解根本 对于这些疾病的病因和有效的治疗方法，我们必须推进我们的 了解调节粘液清除的基本机制。为此，我们的实验室已经 开发了一系列新技术来直接量化一些关键的生化和生物物理 粘液层的特性，包括粘蛋白浓度、粘液渗透压、粘液 粘附力/粘聚力、粘液粘性和弹性模数。我们相信，新的方法可以 量化呼吸道表层的生物物理过程对于弄清黏液清除失败的原因至关重要。 Cf并确定最佳药剂组合(S)以恢复/加速 纤维细胞瘤患者的粘液清除情况。鉴于粘液层在肺健康中的重要性及其变化 在疾病方面，UNC粘液生物化学和生物物理核心的首要目标是提供核心 用户能够评估粘液的各种新的生化和生物物理特性。如中所述 这项提议，核心将允许其他研究人员访问我们的专门粘液/粘蛋白分析，包括 用高效液相色谱/光散射法和折射法测量粘蛋白的绝对浓度； 用免疫学和质谱学方法直接定量粘蛋白；3)胞外DNA 浓度，据称是CF疾病进展的贡献者；4)粘液的重量测量 固体百分比；5)粘液总渗透压和部分渗透压；6)粘液/细胞表面粘附力；7)粘液 粘合强度；8)大块粘液流变学；9)小球流变学；10)纤毛介导率 粘液清除；11)咳嗽介导的粘液清除的比率。UNC粘液筛选核心是 独一无二地定位于：1)表征用户提供的粘液样本的特性以及2) 评估研究人员提供的候选治疗药物对这些生化/生物物理的影响 属性。这一核心的优势是许多科学家的合作，他们是该领域的专家 粘液/粘蛋白分析。对潜在核心用户的好处包括来自核心中心调查人员的建议，如 如何回答粘液特定的问题，以及利用我们的新技术生成数据的能力。这个 北卡罗来纳大学粘液生物化学和生物物理学核心的最终目标是将个人研究人员解放出来 聚焦基础科学和药物开发。

Abnormal mucus clearance is an important contributor to the pathological phenotype of cystic fibrosis (CF). Our studies over the last decade have revealed that CF-related dysfunction of CFTR results in the dehydration of the airway mucus layer. The outcome of such dehydration is significant alterations in the biophysical properties of the mucus layer and its interaction with the cell surface, i.e. producing adherent mucus that sticks to epithelial cells and a slowing/failure of cilia- and cough-mediated mucus clearance. To understand the root causes of the pathology and develop effective therapies to treat such diseases, we must advance our knowledge of the fundamental mechanisms regulating mucus clearance. To this end, our laboratories have developed a series novel techniques to directly quantify a number of key biochemical and biophysical prosperities of the mucus layer, including mucin concentration, mucus osmotic pressure, mucus adhesion/cohesion strength, and mucus viscous and elastic moduli. We believe that novel approaches to quantifying biophysical processes in airway surface layer are critical to discern why mucus clearance fails in CF and to identify the optimal combinations of pharmacological agent(s) to restore/accelerate the rate of mucus clearance in patients with CF. Given the importance of the mucus layer in lung heath and its alteration in disease, the overarching goal of this the UNC Mucus Biochemistry and Biophysics Core is to provide Core users the ability to assess a variety of novel biochemical and biophysical properties of mucus. As detailed in this proposal, the Core will offer other investigators access to our specialized mucus/mucin assay, including measurements of: 1) absolute mucin concentrations by HPLC/light scattering and refractometry methods; 2) direct quantitation of mucins using immunologic and mass spectrometry approaches; 3) extracellular DNA concentration, a purported contributor of disease progression in CF; 4) gravimetric measurements of mucus percent solids; 5) total and partial mucus osmotic pressure; 6) mucus/cell surface adhesion forces; 7) mucus cohesive strength; 8) bulk mucus rheology; 9) small-scale micro-bead rheology; 10) rates of cilia-mediated mucus clearance; and 11) rates of cough-mediated mucus clearance. The UNC mucus screening core is uniquely positioned to both: 1) characterize the properties of user supplied mucus samples as well as 2) assess the impact of investigator supplied candidate therapeutic agents on these biochemical/biophysical properties. The strength of this Core is the collaboration of a number of scientists who are experts in the field of mucus/mucin analysis. The benefits to potential Core users include advice from Core Center Investigators as to how to answer mucus specific questions and the ability to generate data utilizing our novel techniques. The ultimate goal of the UNC Mucus Biochemistry and Biophysics Core is to free up individual investigators to focus on basic science and drug development.