Mucus Biochemistry and Biophysics Core

异常粘液清除是囊性纤维化（CF）病理表型的重要贡献者。 我们在过去十年的研究表明，CF相关的CFTR功能障碍导致脱水， 呼吸道粘液层。这种脱水的结果是生物物理特性的显著改变。 粘液层的性质及其与细胞表面的相互作用，即产生粘附的粘液 以及纤毛和咳嗽介导的粘液清除的减慢/失败。为了了解根源 我们必须研究病理学的原因，并开发治疗这些疾病的有效疗法， 了解调节粘液清除的基本机制。为此，我们的实验室 开发了一系列新的技术，直接量化一些关键的生化和生物物理 粘液层的性质，包括粘蛋白浓度、粘液渗透压、粘液 粘附/内聚强度以及粘液粘性和弹性模量。我们认为，新的方法， 量化气道表面层中的生物物理过程对于辨别为什么粘液清除失败是至关重要的， CF，并确定药理学药物的最佳组合，以恢复/加速 CF患者的粘液清除率。鉴于粘液层在肺健康中的重要性及其变化 在疾病中，北卡罗来纳大学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物化学和生物物理特性。详见 根据这一提议，核心将为其他研究人员提供我们专门的粘液/粘蛋白检测，包括 测量：1）通过HPLC/光散射和折射法的绝对粘蛋白浓度; 2） 使用免疫学和质谱法直接定量粘蛋白; 3）细胞外DNA 浓度，CF中疾病进展的据称贡献者; 4）粘液的重量测量 固体百分比; 5）总粘液渗透压和部分粘液渗透压; 6）粘液/细胞表面粘附力; 7）粘液 内聚强度; 8）大量粘液流变学; 9）小规模微珠流变学; 10）纤毛介导的 粘液清除率;和11）咳嗽介导的粘液清除率。粘液筛选核心， 独特地定位于：1）表征用户提供的粘液样品的性质以及2） 评估研究者提供的候选治疗药物对这些生化/生物物理学的影响 特性.这个核心的力量是一些科学家的合作，他们是该领域的专家， 粘液/粘蛋白分析。潜在核心用户的受益包括核心中心研究者的建议， 如何回答粘液的具体问题，以及利用我们的新技术生成数据的能力。的 粘液生物化学和生物物理学核心的最终目标是解放个体研究者， 专注于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.