Mucolytic for enhanced mucus and biofilm clearance in cystic fibrosis patients

描述(申请人提供)：粘稠的粘液和延迟的粘液纤毛清除是囊性纤维化(CF)发病机制的重要因素，但目前可用的治疗方法对对抗粘液淤积的疗效有限。此外，填充在肺组织中的生物膜会降低肺功能，通常不会受到当前粘液类药物的影响，并严重限制了抗菌效果。新型黏液溶解疗法可以降低痰中CF的黏度，降低病原菌生物膜的活力、粘附力和黏度，改善黏液纤毛清除，将为对抗呼吸衰竭提供重要的治疗进展。我们的初步数据表明，PAAG是一种可溶性、无毒的聚阳离子多糖，对CF痰有很强的抑制作用，并能降低从CF分离株生长的生物膜的粘度和凝聚力。生物膜凝聚力的降低预计将增强标准抗生素的活性。在此基础上，我们假设PAAG可能是一种有效的肺黏液溶解剂，可以改善CF患者的呼吸道清除和提高抗生素活性，但需要进一步的研究来了解其药理、配方和广泛的作用范围，以建议开发为人类治疗药物。PAAG活性的展示将提供至关重要的概念数据证据，并为在人类受试者中测试其活性提供科学依据。在这个项目中，我们将评估PAAG的 使用最先进的流变仪降低患者痰的粘度的作用。此外，我们将通过评估纤毛细胞单层上的活性来证明粘度的降低改善了粘液纤毛的清除。我们还将通过更好地再现生理条件的标准生物膜筛选试验和流动细胞试验，确定适当的剂量和处理方案，以降低生物膜的凝聚力。生物被膜的连贯性降低也将与标准的抗生素治疗相结合，以了解PAAG对用于CF患者的治疗的增强作用。项目完成后，使用PAAG进行进一步开发，将证明PAAG是一种新的、高效的CF粘液溶解疗法，以满足CF患者护理中未得到满足的需求。

DESCRIPTION (provided by applicant): Thick hyperviscous mucus and delayed mucociliary clearance are important elements underlying cystic fibrosis (CF) pathogenesis, but currently available therapeutics exhibit limited efficacy to combat mucus stasis. In addition, biofilms that populate the CF lung contribute to reduced lung function and are typically not affected by current mucolytics and severely limit antimicrobial efficacy. Novel mucolytic therapies that innately reduce the viscosity of CF sputum, reduce the viability, cohesion and viscosity of pathogenic biofilms and improve mucociliary clearance would provide an important therapeutic advance to combat respiratory decline. Our preliminary data demonstrate that PAAG, a soluble, nontoxic polycationic polysaccharide, has robust effects on CF sputum and reduces the viscosity and cohesion of biofilms grown from CF isolates. This reduction in biofilm cohesion is anticipated to enhance activity of standard antibiotics. Based on this, we hypothesize that PAAG could be an effective pulmonary mucolytic for CF patients that will improve airway clearance and improve antibiotic activity, but requires further studies to understand its pharmacology, formulation, and breadth of effects to propose development as a human therapeutic. Demonstration of PAAG's activity will provide crucial proof of concept data and provide a scientific basis for testing its activity in human subjects. In this project, we will assess PAAG's role in reducing the viscosity of patient derived sputum using state-of-the-art rheometry. Further we will demonstrate that the reduction in viscosity improves mucociliary clearance by assessing activity on monolayers of ciliated cells. We will also determine appropriate dosing and treatment regimes for reducing the cohesion of biofilms by standard biofilms screening assays and flow cells assays that better reproduce physiologic conditions. The reduced coherence of biofilms will also be combined with standard antibiotic treatments to understand the potentiating effects of PAAG on therapies used in CF patients. Upon completion of the project, the use of PAAG for further development will have proof of concept as a new and highly effective CF mucolytic treatment to address an unmet need in CF patient care.