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

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.

项目概要有效的哮喘管理需要调节气道平滑肌 (ASM) 收缩状态以避免或是否通过使用直接支气管扩张剂（例如β-激动剂）来尝试逆转支气管收缩。抗炎药（例如皮质类固醇）或两者的某种组合，往往缺乏管理，据估计，55% 的哮喘患者控制效果不佳，目前所有支气管扩张药物均存在局限性。最常用的支气管扩张剂类别的功效和安全性问题仍然存在我们认为，支气管扩张药物的局限性是可以克服的。通过针对 ASM 收缩状态 2 个最强大的调节因子的方法：药物力学我们追求β-激动剂的组合（主要靶向）。药物机械耦合）和专门针对肌动蛋白细胞骨架的药物可能非常有效支气管扩张剂，具有功能协同作用，可降低药物剂量，从而提高安全性在目标 1 中，我们将使用细胞、组织和体内来建立三个目标。 ASM 收缩模型、药物机械耦合联合靶向的协作性质和肌动蛋白聚合，以及 β 激动剂和细胞骨架靶向药物的最佳组合在目标 2 中，我们将通过表征来确定这种功能协同性的机制基础。这些药物对控制跨桥循环的信号中间体和结果的影响（肌球蛋白轻链激酶和磷酸酶磷酸化）或肌动蛋白聚合状态（F/G肌动蛋白比），以及最后，在目标 3 中，我们将评估哮喘病理学的影响。 β-激动剂和肌动蛋白细胞骨架靶向药物联合使用抑制 ASM 收缩的功效气道阻力，通过采用源自人类细胞/组织的细胞和组织模型系统哮喘患者，或其中哮喘病理学被施加在体外/离体（细胞、组织）或体内（2 不同的体内小鼠模型）。拟议的研究由 3 个已建立的具有互补性的 PI 进行。专业知识代表了建立哮喘管理策略的创新方法，该策略克服了此外，拟议的机制研究将提供新的见解。研究如何以最佳方式破坏跨桥循环和细胞骨架硬化之间的合作在 ASM 细胞中产生张力。