Phosphorylation of ciliary target proteins

纤毛靶蛋白的磷酸化

基本信息

批准号：
8039271
负责人：
Matthias A Salathe
金额：
$ 38.25万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-07-01 至 2013-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8039271
关键词：
Address Adenylate Cyclase Adherent Culture Adrenergic Agonists Affect Agonist Albuterol Arthritis Asthma Bicarbonates Biological Assay Blood Vessels Breathing Bronchiectasis Bronchodilation Bronchodilator Agents Calcium Carbon Dioxide Carnitine Cations Cell Line Cell physiology Cells Charge Chronic Obstructive Airway Disease Cilia Clinical Cyclic AMP Data Diffuse Disease Drug Transport Environment Enzymes Epithelial Epithelial Cells Exhalation Fluorescence Resonance Energy Transfer Forskolin Frequencies Functional disorder GTP-Binding Proteins Goals Health Height Homeostasis Human Human Volunteers Individual Inflammation Joints Lead Liquid substance Measures Mediating Molecular Mucociliary Clearance Mucous Membrane NADPH Oxidase Pathway interactions Pharmaceutical Preparations Phosphorylation Physiological Play Production Proteins Regulation Role Second Messenger Systems Small Interfering RNA Smooth Muscle Societies Surface Testing Time Tissues absorption airway epithelium airway inflammation airway surface liquid base clinically relevant cytokine extracellular improved in vivo inhibitor/antagonist new therapeutic target novel prevent second messenger uptake

项目摘要

DESCRIPTION (provided by applicant): In airway disease exacerbations, the airway acidifies as measured by low pH in exhaled breath condensate in asthma, COPD and bronchiectasis. Clinically, ¿-adrenergic agonists are used to increase cAMP via stimulation of transmembrane adenylyl cyclases (tmACs), with the intent to enhance mucociliary transport and bronchodilation during disease exacerbations. For bronchodilation, inhaled drugs need to cross the airway epithelium. The majority of the currently used bronchodilators have a positive charge at physiological or acidic pH. Thus, these drugs cannot freely diffuse across the airway epithelium to reach their targets. We have shown that the pH-dependent organic cation/carnitine transporters OCTN1 and OCTN2 are expressed in airway epithelia and that they transport cations, including albuterol, into airway epithelial cells in a pH-dependent manner. Uptake of positively charged bronchodilators is decreased in an acidic airway environment, suggesting that their bronchodilatory effects will be reduced. We have also shown that intracellular acidification directly inhibits ciliary beating. As a counter measure, increases in intracellular HCO3- stimulate cilia via a pathway involving cAMP production by a novel, cytosolic (or soluble) adenylyl cyclase (sAC). These new findings suggest a significant role for pHi and HCO3- in the regulation of human airway homeostasis. We hypothesize that pH directly and HCO3-, likely through sAC, play critical roles in regulating airway epithelial cell functions, including uptake of charged cations such as bronchodilators. This hypothesis will be comprehensively tested with three specific aims, including a bioassay that assesses the ability of inhaled albuterol to dilate airway blood vessels in normal human volunteers. Aim 1 will test the hypothesis that cAMP production is accomplished by both tmACs and sAC and thus influenced by changes in HCO3-, pH, Ca2+. Aim 2 will test the hypothesis that pH-dependent cation uptake into airway epithelial cells will make ¿-adrenergic agonists available in higher concentrations on the airway surface during acidification. Aim 3 will test the hypothesis that airway acidification inhibits transepithelial cation uptake, including albuterol, into airway submucosal tissues in human beings. This application thus addresses a novel and clinically relevant regulation mechanism of mucociliary clearance and of drug availability on the epithelial surface as well as in the subepithelial tissue using a translational and state-of-the-art approach. PUBLIC HEALTH RELEVANCE. Airway diseases and specifically their exacerbations pose important health problems in the USA and worldwide. The airway acidifies during disease exacerbations and this change in pH is predicted to decrease uptake of bronchodilators across the airway epithelium to their target (namely smooth muscle) and adversely affect aspects of mucociliary clearance. This application examines mechanisms by which bronchodilators are taken up into the airway mucosa and novel aspects of mucociliary clearance regulation. Improving ways that bronchodilators reach their target during disease exacerbations and preventing mucociliary dysfunction or restoring clearance to normal levels is the ultimate clinical goal. This proposal thus may identify new therapeutic targets that could help to decrease the burden from airway disease exacerbations on individuals and on society.

描述（由申请人提供）：在气道疾病加剧中，气道通过低pH值在哮喘，COPD和支气管扩张中的呼气呼吸冷凝物中酸化。在临床上，肾上腺素激动剂用于通过刺激跨膜腺苷酸周期（TMAC）来增加cAMP，目的是增强疾病加剧期间的粘膜纤毛转运和支气管扩张。对于支气管扩张，遗传药物需要越过气道上皮。当前使用的大多数支气管扩张剂在生理或酸性pH值时具有正电荷。那就是这些药物不能在气道上皮上自由扩散以达到目标。我们已经表明，pH依赖性有机阳离子/肉碱转运蛋白octn1和octn2在气道上皮中表达，它们以pH值依赖性方式将阳离子（包括pleosol）运输到气道上皮细胞中。在酸性气道环境中，带正电的支气管扩张剂的吸收减少，表明其支气管扩张剂将减少。我们还表明，细胞内酸化直接抑制纤毛跳动。作为计数器测量，细胞内HCO3的增加通过涉及cAMP生产的途径刺激纤毛，这是一种新型的胞质（或固体）腺苷酸环化酶（SAC）。这些新发现表明，PHI和HCO3-在人类气道体内平衡的调节中起着重要作用。我们假设pH值直接通过SAC，可以通过SAC来控制气道上皮细胞功能，包括吸收带电阳离子（例如支气管扩张剂）。该假设将通过三个特定目的对这一假设进行全面检验，包括一项生物测定法，该生物测定法评估了掺入倍醇在正常人类志愿者中扩张气道血管扩张的能力。 AIM 1将检验以下假设：CAMP生产是由TMAC和SAC完成的，因此受HCO3-，pH，Ca2+的变化的影响。 AIM 2将检验以下假设：pH依赖性阳离子摄取对气道上皮细胞的摄取将使酸化过程中较高浓度的肾上腺肾上腺激动剂可用。 AIM 3将检验以下假设：气道酸化抑制了包括铜酚在内的换皮阳离子吸收，包括人类的气道粘膜下组织。因此，使用转化和最先进的方法，该应用解决了上皮表面以及上皮组织中的新型和临床相关的调节机制。公共卫生相关性。气道疾病及其加剧的疾病在美国和全球构成了重要的健康问题。疾病加剧期间的气道酸化，预计pH值的这种变化将减少跨气道上皮的支气管扩张剂对其靶标（即平滑肌）的摄取，并对粘膜清除率的各个方面产生不利影响。这种应用检查机制，通过将支气管扩张剂吸收到气道粘膜和粘膜纤毛清除调节的新方面。在疾病加剧期间，支气管扩张剂达到目标并防止粘膜助力功能障碍或将清除恢复到正常水平的方法是最终的临床目标。因此，该建议可能会确定新的治疗靶标，这些靶标有助于减少气道疾病对个人和社会的加剧。

项目成果

期刊论文数量（29）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Soluble adenylyl cyclase is localized to cilia and contributes to ciliary beat frequency regulation via production of cAMP.

DOI：
10.1085/jgp.200709784
发表时间：
2007-07
期刊：
The Journal of general physiology
影响因子：
0
作者：
Schmid A;Sutto Z;Nlend MC;Horvath G;Schmid N;Buck J;Levin LR;Conner GE;Fregien N;Salathe M
通讯作者：
Salathe M

Effect of airway acidosis and alkalosis on airway vascular smooth muscle responsiveness to albuterol.

DOI：
10.1186/s40360-015-0008-y
发表时间：
2015-04-02
期刊：
BMC pharmacology & toxicology
影响因子：
2.9
作者：
Cancado JE;Mendes ES;Arana J;Horvath G;Monzon ME;Salathe M;Wanner A
通讯作者：
Wanner A

Soluble adenylyl cyclase in health and disease.

DOI：
10.1016/j.bbadis.2014.07.010
发表时间：
2014-12
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE
影响因子：
6.2
作者：
Schmid, Andreas;Meili, Dimirela;Salathe, Matthias
通讯作者：
Salathe, Matthias

Calcium-mediated, purinergic stimulation and polarized localization of calcium-sensitive adenylyl cyclase isoforms in human airway epithelia.

人气道上皮细胞中钙介导的嘌呤能刺激和钙敏感腺苷酸环化酶亚型的极化定位。

DOI：
10.1016/j.febslet.2007.06.015
发表时间：
2007
期刊：
FEBS letters
影响因子：
3.5
作者：
Nlend,Marie-Christine;Schmid,Andreas;Sutto,Zoltan;Ransford,GeorgeA;Conner,GregoryE;Fregien,Nevis;Salathe,Matthias
通讯作者：
Salathe,Matthias

Post-secretory fate of host defence components in mucus.

粘液中宿主防御成分的分泌后命运。