Cooperative targeting of pharmacomechanical coupling and the actin cytoskeleton to regulate ASM contraction

药物机械耦合和肌动蛋白细胞骨架的协同靶向调节 ASM 收缩

基本信息

批准号：
10434061
负责人：
RAMASWAMY KRISHNAN
金额：
$ 46.57万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10434061
关键词：
ABL1 gene Acetylcholine Actins Adrenal Cortex Hormones Agonist Air Movements Airway Resistance Anti-Inflammatory Agents Asthma Biological Models Bronchoconstriction Bronchodilator Agents Cell model Cells Chronic Combined Modality Therapy Coupling Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cytoskeleton Development Dose Drug Targeting Drug usage Fourier Transform G Actin G-Protein-Coupled Receptors Goals Human In Vitro Interleukin-13 LIM Domain Kinase 1 Leukotriene Antagonists Lung Mediating Microscopy Modeling Morbidity - disease rate Mus Muscarinics Muscle Contraction Muscle Tension Muscle relaxants Muscle relaxation phase Myosin Light Chain Kinase Myosin Light Chains Nature Outcome Pharmaceutical Preparations Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Receptor Activation Receptor Signaling Regulation Reporting Safety Signal Transduction Slice Smooth Muscle Myocytes Structure of parenchyma of lung TNF gene Testing Therapeutic Tissue Model Tissues Traction Wheezing antagonist asthmatic compare effectiveness constriction cysteinyl leukotriene receptor human tissue in vivo in vivo Model inhibitor innovation insight mortality mouse model myosin phosphatase polymerization respiratory smooth muscle

ABL1 gene Acetylcholine Actins Adrenal Cortex Hormones Agonist Air Movements Airway Resistance Anti-Inflammatory Agents Asthma Biological Models Bronchoconstriction Bronchodilator Agents Cell model Cells Chronic Combined Modality Therapy Coupling Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cytoskeleton Development Dose Drug Targeting Drug usage Fourier Transform G Actin G-Protein-Coupled Receptors Goals Human In Vitro Interleukin-13 LIM Domain Kinase 1 Leukotriene Antagonists Lung Mediating Microscopy Modeling Morbidity - disease rate Mus Muscarinics Muscle Contraction Muscle Tension Muscle relaxants Muscle relaxation phase Myosin Light Chain Kinase Myosin Light Chains Nature Outcome Pharmaceutical Preparations Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Receptor Activation Receptor Signaling Regulation Reporting Safety Signal Transduction Slice Smooth Muscle Myocytes Structure of parenchyma of lung TNF gene Testing Therapeutic Tissue Model Tissues Traction Wheezing antagonist asthmatic compare effectiveness constriction cysteinyl leukotriene receptor human tissue in vivo in vivo Model inhibitor innovation insight mortality mouse model myosin phosphatase polymerization respiratory smooth muscle

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项目摘要

Project Summary Effective asthma management requires regulating airway smooth muscle (ASM) contractile state to avoid or reverse bronchoconstriction. Whether this is attempted by use of direct bronchodilators (e.g., β-agonists), by anti-inflammatory agents (e.g., corticosteroids), or some combination of both, too often management is lacking, as an estimated 55% of all asthmatics have suboptimal control. All current bronchodilator drugs have limitations which respect to efficacy, and safety issues still persist with the most frequently used class of bronchodilator drugs- long-acting β-agonists (LABAs). We submit that the limitations of bronchodilator drugs can be overcome by an approach that targets the 2 most powerful regulators of ASM contractile state: pharmacomechanical coupling and the actin cytoskeleton. We hypothesize that combinations of beta-agonists (that primarily target pharmacomechanical coupling) and drugs that specifically target the actin cytoskeleton can be highly efficacious bronchodilators, with a functional cooperatively that enables lower drug doses and therefore a better safety profile. Three aims are proposed to test this hypothesis. In Aim 1, we will establish, using cell, tissue, and in vivo models of ASM contraction, the cooperative nature of combined targeting of pharmacomechanical coupling and actin polymerization, and identify optimal combinations of beta-agonists and cytoskeleton-targeting drugs that relax ASM. In Aim 2, we will determine the mechanistic basis for this functional cooperativity by characterizing the effects of these drugs on signaling intermediates and outcomes that control cross bridge cycle (myosin light chain kinase and phosphatase phosphorylation) or actin polymerization state (F/G actin ratio), and on the upstream signals that regulate these outcomes. Lastly, in Aim 3 we will assess the effect of asthma pathobiology on the efficacy of combining β-agonists and actin cytoskeleton-targeting drugs in inhibiting ASM contraction and airway resistance, by employing cell and tissue model systems derived from cells/tissues from human asthmatics, or in which asthma pathobiology is imposed either in vitro/ex vivo (to cells, tissue), or in vivo (2 differrent in vivo murine models). The proposed studies performed by 3 established PIs with complementary expertise represent an innovative approach to establish an asthma management strategy that overcomes the current limitations of efficacy and safety. Moreover, the proposed mechanistic studies will provide new insight into how to optimally disrupt the cooperation between cross bridge cycling and cytoskeleton stiffening that generates tension in the ASM cell.

项目摘要有效的哮喘管理需要调节气道平滑肌（ASM）收缩状态才能避免或反向支气管收缩。是否通过使用直接支气管扩张剂（例如β-激动剂）来尝试这抗炎剂（例如皮质类固醇）或两者的某种组合，经常缺乏管理，据估计，所有哮喘患者中有55％具有次优控制。所有当前的支气管扩张药都有局限性关于效率和安全性问题的问题仍然与最常用的支气管扩张剂相关药物 - 长效β-激动剂（LABAS）。我们认为可以克服支气管扩张药的局限性通过针对ASM收缩状态的两个最强大调节器的方法：药物力学耦合和肌动蛋白细胞骨架。我们假设β-激动剂的组合（该主要目标专门针对肌动蛋白细胞骨架的药物力学耦合）和药物效率很高支气管扩张剂，在功能合作上可以使药物剂量降低，因此可以更好地安全轮廓。提出了三个目的来检验这一假设。在AIM 1中，我们将建立，使用细胞，组织和体内 ASM收缩的模型，药物力学耦合和合并靶向的合作性质和肌动蛋白聚合，并确定β-激动剂和细胞骨架靶向药物的最佳组合放松ASM。在AIM 2中，我们将通过表征该功能协调的机械基础这些药物对控制跨桥周期（肌球蛋白光）的信号中间体和结果的影响链激酶和磷酸酶磷酸化）或肌动蛋白聚合态（F/G肌动蛋白比），并在调节这些结果的上游信号。最后，在AIM 3中，我们将评估哮喘病理生物学的影响关于结合β-激动剂和肌动蛋白细胞骨架靶向药物在抑制ASM收缩和通过使用源自人类的细胞/组织的细胞和组织模型系统，气道电阻哮喘患者或哮喘病理生物学是在体外/外体内施加的（细胞，组织）或体内的（2 不同的体内鼠模型）。由3个建立的PI进行完整的PIS进行的拟议研究专业知识代表了一种创新的方法来建立哮喘管理策略，以克服当前的效率和安全性局限性。此外，拟议的机械研究将提供新的见解如何最佳破坏跨桥骑自行车与细胞骨架之间的合作在ASM单元中产生张力。