Targeting airway smooth muscle chloride fluxes for bronchorelaxation

靶向气道平滑肌氯化物通量以实现支气管舒张

基本信息

批准号：
9054914
负责人：
CHARLES W EMALA
金额：
$ 40万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-15 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9054914
关键词：
Acetylcholine Acute Aerosols Affect Agonist Anti-Inflammatory Agents Anti-inflammatory Asthma Breathing Bronchoconstriction CLCA2 gene Calcium Cavia Cell membrane Cells Chloride Channels Chloride Ion Chlorides Chronic Clinical Clinical Research Country Coupling Data Disease Drug Targeting Effectiveness Epithelial Family Figs - dietary Fluorescence Resonance Energy Transfer Generations Genetic Histamine Human Image Incidence Inflammation Inhalation Therapy Inositol Phosphates Left Leukotrienes Libraries Ligands Link Literature Lung MUC5AC gene Mediating Mediator of activation protein Membrane Potentials Modeling Molecular Mucous body substance Mus Muscle Tonus Muscle relaxation phase Nerve Niflumic Acid Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Population Potassium Chloride Prevalence Prevention Prodrugs Production Pyroglyphidae Relaxation Reporting Resistance Role Safety Sarcoplasmic Reticulum Signal Pathway Smooth Muscle Smooth Muscle Myocytes Sodium Specificity Text Therapeutic Time aerosolized airway epithelium airway inflammation airway obstruction base cell type clinically relevant constriction cost drug discovery experience extracellular genetic regulatory protein in vivo inhibitor/antagonist mortality mouse model novel novel therapeutic intervention novel therapeutics prevent public health relevance relating to nervous system respiratory smooth muscle statistics symporter systemic toxicity voltage

项目摘要

DESCRIPTION (provided by applicant): Over 300 million people in the world suffer from asthma. The US is among the countries with the highest incidence (10.9% of the population) with a yearly cost of $56 billion. Despite these staggering statistics, there have been no new classes of medications targeting airway smooth muscle (ASM)-mediated bronchoconstriction in asthma for many decades. Moreover, long-acting -agonists, the current leading therapy that directly targets ASM constriction, have been associated with an increased mortality from asthma and are currently under FDA-mandated safety review in 5 clinical studies. Our preliminary studies support the hypothesis that drugs targeting the channels and transporters that control chloride flux in ASM may be novel therapies for relaxing ASM. We propose functional and mechanistic studies to document the efficacy of targeting calcium-activated chloride channels alone or in combination with sodium-potassium-chloride co-transporters as novel asthma therapies. We propose that inhaled therapy will circumvent concerns related to systemic toxicity and will confirm existing literature that blockade of these channels also reduces airway mucus production. Thus, we propose the following 3 specific aims: Aim 1a: functional relaxation of ex vivo human and guinea pig ASM by targeting chloride flux pathways. We will demonstrate the effectiveness of blockade of chloride channels alone or in combination with chloride transporters in the prevention of contraction or the induction of relaxation. We will also determine the ability of chloride channel/transporter blockade to potentiate relaxation of 2- agonists in human ASM providing initial translational data for clinically relevant drug discovery. Aim 2: functional reduction of in vivo lung resistance. We will demonstrate that acute aerosol delivery of blockers of chloride channels +/- chloride transporters prevents bronchoconstriction in vivo in 3 mouse models: (1) naively hyperresponsive A/J strain (2) house dust mite sensitized C57 mice, and (3) a mouse with a selective genetic deletion of the TMEM16A chloride channel in smooth muscle. The effect on epithelial mucous production will be assessed in these models. Aim 3: the cellular mechanism(s) by which chloride channels regulate ASM tone. We will demonstrate the link between chloride control of plasma membrane (PM) potential and intracellular signaling pathways linked to contraction/relaxation including stored operated Ca2+ entry, Gq-coupling to inositol phosphate generation, and phosphorylation of contractile-regulatory proteins (RhoA, MYPT1, MLC). We will distinguish between chloride control of Ca2+ flux across the PM vs SR using a FRET-based SR-specific Ca2+ indicator (D1ER).

描述（由申请人提供）：美国是哮喘发病率最高的国家之一（占人口的 10.9%），每年造成的费用高达 560 亿美元。几十年来，一直没有针对哮喘气道平滑肌（ASM）介导的支气管收缩的新药物。此外，长效β受体激动剂是目前直接针对ASM收缩的主要疗法。与哮喘死亡率增加有关，目前正在进行 FDA 授权的 5 项临床研究安全审查，我们的初步研究支持这样的假设：针对控制 ASM 中氯离子流量的通道和转运蛋白的药物可能是缓解 ASM 的新疗法。我们建议进行功能和机制研究，以记录单独靶向钙激活氯离子通道或与钠钾氯协同转运蛋白联合作为新型哮喘疗法的功效。将证实现有文献表明，封锁这些通道也可以减少气道粘液的产生。因此，我们提出以下 3 个具体目标：目标 1a：通过针对氯化物通量途径来放松体外人类和豚鼠 ASM 的功能。我们还将确定单独阻断氯离子通道或与氯离子转运蛋白组合来预防收缩或诱导松弛的能力。氯离子通道/转运蛋白阻断以增强人 ASM 中 2- 激动剂的松弛，为临床相关药物发现提供初步转化数据目标 2：体内肺阻力的功能性降低我们将证明氯离子通道阻断剂的急性气雾输送 +。 /- 氯离子转运蛋白在 3 种小鼠模型中预防体内支气管收缩：(1) 幼稚高反应性 A/J 品系 (2) 屋尘螨致敏的 C57 小鼠，以及 (3) 平滑肌中选择性遗传删除 TMEM16A 氯离子通道的小鼠将在这些模型中评估对上皮粘液产生的影响：氯离子通道的细胞机制。我们将证明质膜 (PM) 电位的氯化物控制与收缩/舒张相关的细胞内信号通路（包括储存操作的 Ca2+ 进入）之间的联系。 Gq 偶联肌醇磷酸盐的生成以及收缩调节蛋白（RhoA、MYPT1、MLC）的磷酸化我们将使用基于 FRET 的 SR 特异性 Ca2+ 指示剂 (D1ER) 来区分 PM 和 SR 中 Ca2+ 通量的氯化物控制。。