A Ginsenoside TMEM16A Potentiator for Cystic Fibrosis

项目总结 囊性纤维化跨膜电导调节器(CFTR)氯离子通道功能障碍阻碍粘液 清除并导致呼吸道疾病，囊性纤维化(CF)。新型CFTR调节剂改善粘液纤毛 基因突变特异性患者的运输(MCT)和疾病的病理生理表现，但 不能完全恢复呼吸道功能。我们最近发现了从朝鲜红中提取的人参皂苷 人参作为钙激活的Cl-−通道[TMEM16A]的增强剂，可使MCT恢复 体外和体内通过这一替代的氯离子途径。然而，TMEM16A激活剂的临床试验结果不佳 最近被描述的通道在受感染的呼吸道粘液高分泌中的作用导致了一些 提示抑制TMEM16A是一种更好的策略。然而，我们假设增强TMEM16A， 不是通过嘌呤能的通道激活，而是细胞内钙离子驱动的通路，将挽救MCT 感染的CF型呼吸道。特定目标1将分离最有效地增强TMEM16A和TMEM16A的人参皂苷 测定其对MCT参数和粘液病理的影响。我们期待找到最佳的人参皂苷 通过Ussing小室和膜片钳实验，同时测量对MCT标志物的影响 和粘液粘度。特定目标2将评估TMEM16A增强剂是否改善上皮细胞氯- 感染的CF大鼠的呼吸道分泌物、组织病理学和MCT参数。我们将确定单次剂量 人参皂苷在假单胞菌感染的CF大鼠体内的药代动力学研究 鼻电位差，MCT，肺微CT，组织病理学，粘液特性(黏度，固体，质量， 密度)和炎性细胞因子。我们将回答有关靶向治疗效用的问题 TMEM16A途径，并确定一种新的TMEM16A增强剂作为治疗CF呼吸道疾病的有效药物。

PROJECT SUMMARY Dysfunction of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel impedes mucus clearance and leads to the airway disease, cystic fibrosis (CF). New CFTR modulators improve mucociliary transport (MCT) and pathophysiologic manifestations of the disease in genetic mutation-specific patients, but do not completely restore airway function. We recently discovered ginsenosides derived from Korean Red Ginseng serve as potentiators of the Ca2+-activated Cl− channel (CaCC) [TMEM16A] and can restore MCT in vitro and in vivo through this alternative Cl- pathway. Yet, poor clinical trial results with activators of TMEM16A and a recently described role for the channel in mucus hypersecretion in infected airways has led to some to suggest that inhibiting TMEM16A is a better strategy. However, we hypothesize that potentiating TMEM16A, rather than channel activation through purinergic, intracellular Ca2+- driven pathways, will rescue MCT in infected CF airways. Specific Aim 1 will isolate the ginsenoside that most effectively potentiates TMEM16A and measure its effects on MCT parameters and mucus pathology. We anticipate finding the optimal ginsenoside through Ussing chamber and patch clamp experiments, while also measuring the impact on markers of MCT and mucus viscosity. Specific Aim 2 will assess whether a TMEM16A potentiator improves epithelial Cl- secretion, histopathology, and MCT parameters in infected CF rat airways. We will determine single dose pharmacokinetics for the selected ginsenoside and administer to Pseudomonas-infected CF rats to evaluate nasal potential difference, MCT, lung micro-CTs, histopathology, mucus properties (viscosity, solids, mass, density), and inflammatory cytokines. We will answer questions about the therapeutic utility of targeting the TMEM16A pathway and identify a new TMEM16A potentiator as an effective treatment for CF airway disease.