Inhibition of TMEM16 proteins to attenuate mucus secretion and to improve airway clearance

抑制 TMEM16 蛋白可减少粘液分泌并改善气道清除

• 批准号: 407638684
• 负责人: Professor Dr. Karl Kunzelmann
• 金额: --
• 依托单位: Institut für Physiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/407638684?language=en
• 关键词: Inhibition; TMEM16; proteins; attenuate; mucus

A central problem in asthma and cystic fibrosis (CF) is the accumulation of large amounts of mucus in the airways. In CF, the mucus is particularly viscous, adhesive, and appears dehydrated, leading to mucus obstructions in airways and intestine. Chronic inflammation causes goblet cell metaplasia and upregulation of the calcium activated Cl- channel TMEM16A. It is, however, unclear if TMEM16A is essential for mucus production and/or secretion. In addition, it is unknown whether upregulation and activation of TMEM16A is beneficial for mucociliary clearance or whether this contributes to impaired airway clearance. Our published and preliminary (unpublished) results from tissue specific TMEM16A and TMEM16F knockout mice suggest that TMEM16A is essential for mucus release, while TMEM16F is relevant for mucus production. TMEM16A and 16F are Cl- channels expressed in mouse and human airways and intestine, and are essential for CFTR to operate. We reported that fluid secretion was essentially absent in TMEM16A-/- mice, yet mucociliary transport was even enhanced. Moreover, inhibition of TMEM16 improved airway function in preliminary experiments with asthmatic mice. Using TMEM16A and 16F knockout mice and pluripotent human airway epithelial cells, we will clarify: i) Whether TMEM16F is essential for mucus production. ii) Whether TMEM16A is indispensable for mucus secretion but unimportant for mucus production. iii) Whether blockade of TMEM16 in asthmatic mice will attenuate mucus secretion, enhance mucociliary clearance, and improve airway function. iv) If activation of TMEM16 has adverse effects in mouse airways. Mucus transport and mucociliary clearance will be measured in mice in vivo and ex vivo in perfused mouse and porcine airways, and in mouse intestine. We will investigate whether the FDA-approved compound and inhibitor of TMEM16, niclosamide ethanolamine, inhibits mucus production and mucus secretion. We will test whether the drug attenuates airway plugging in asthmatic mice and intestinal obstruction in CF mice. Similar experiments will be performed using a novel and highly potent inhibitor from AMGEN (ANO1-InhibA). Using activators of TMEM16 (Eact or C5), we will demonstrate that activation of TMEM16 has a negative impact on airway function. This is the first study that will clearly demonstrate the role of TMEM16 for airway mucus clearance and mucus plugging in inflammatory airway disease. It will provide novel insights into the cellular mechanisms of mucus release. The study is pressing because its results are needed to guide further TMEM16-based strategies to reduce airway mucus plugging in asthma and CF.

哮喘和囊性纤维化 (CF) 的一个核心问题是气道中积聚大量粘液。在 CF 中，粘液特别粘稠、有粘性，并且出现脱水，导致气道和肠道粘液阻塞。慢性炎症会导致杯状细胞化生和钙激活 Cl 通道 TMEM16A 的上调。然而，尚不清楚 TMEM16A 是否对于粘液产生和/或分泌至关重要。此外，尚不清楚 TMEM16A 的上调和激活是否有利于粘膜纤毛清除，或者是否会导致气道清除受损。我们发表的和初步（未发表）的组织特异性 TMEM16A 和 TMEM16F 敲除小鼠的结果表明 TMEM16A 对于粘液释放至关重要，而 TMEM16F 与粘液产生相关。 TMEM16A 和 16F 是在小鼠和人类气道和肠道中表达的 Cl- 通道，对于 CFTR 的运行至关重要。我们报道TMEM16A-/-小鼠基本上不存在液体分泌，但粘液纤毛运输甚至增强。此外，在哮喘小鼠的初步实验中，抑制 TMEM16 可以改善气道功能。使用 TMEM16A 和 16F 敲除小鼠以及多能人气道上皮细胞，我们将阐明： i) TMEM16F 是否对于粘液产生至关重要。 ii) TMEM16A对于粘液分泌是否是必不可少的，但对于粘液产生是否不重要。 iii) 在哮喘小鼠中阻断TMEM16是否会减弱粘液分泌、增强粘液纤毛清除并改善气道功能。 iv) 如果 TMEM16 的激活对小鼠气道产生不利影响。将在小鼠体内和离体灌注小鼠和猪气道以及小鼠肠道中测量粘液转运和粘液纤毛清除率。我们将研究 FDA 批准的 TMEM16 化合物和抑制剂氯硝柳胺乙醇胺是否抑制粘液产生和粘液分泌。我们将测试该药物是否可以减轻哮喘小鼠的气道堵塞和 CF 小鼠的肠梗阻。类似的实验将使用 AMGEN 的新型高效抑制剂（ANO1-InhibA）进行。使用 TMEM16 激活剂（Eact 或 C5），我们将证明 TMEM16 激活对气道功能有负面影响。这是第一项明确证明 TMEM16 在炎症性气道疾病中清除气道粘液和堵塞粘液的作用的研究。它将为粘液释放的细胞机制提供新的见解。这项研究非常紧迫，因为需要其结果来指导进一步基于 TMEM16 的策略，以减少哮喘和 CF 中的气道粘液堵塞。