Targeting T2 inflammation-evoked mechanical endotypes of ASM shortening in asthma

靶向哮喘中 ASM 缩短的 T2 炎症诱发机械内型

• 批准号: 10657988
• 负责人: Steven S An
• 金额: $ 58.28万
• 依托单位: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-25 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657988
• 关键词: Acute; Adjuvant Therapy; Adrenergic Receptor; Adverse effects; Affect; Agonist; Anti-Inflammatory Agents; Asthma; Biological Products; Bronchial Spasm; Bronchodilation; Bronchodilator Agents; Calcium; Cells; Chronic; Clinical; Coupling; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Development; Drug Tolerance; Enhancers; Epigenetic Process; Event; Family; G-Protein-Coupled Receptors; Genetic Transcription; Genomic approach; Goals; Homologous Gene; Human; Immune; Immune response; Immunologics; In Vitro; Inflammation; Inflammatory Response; Interleukin-13; Kinetics; Knowledge; Lung; Mechanics; Mediating; Mediator; Methods; MicroRNAs; Molecular; Mus; Muscle relaxation phase; Obstruction; Obstructive Lung Diseases; Pathogenicity; Patients; Persons; Pharmaceutical Preparations; Physiological; Polycomb; Pre-Clinical Model; Predisposition; Property; Proteins; Psychological reinforcement; Receptor Activation; Receptor Inhibition; Regulation; Regulatory Element; Relaxation; Research; Role; Sensory; Signal Transduction; Slice; Smooth Muscle Myocytes; Tachyphylaxis; Taste Buds; Therapeutic; Tissues; Transcription Repressor; Transforming Growth Factor beta; Translational Repression; Work; airway inflammation; airway obstruction; airway remodeling; clinically relevant; cohort; computer studies; cytokine; desensitization; experimental study; genetic approach; improved; in vivo Model; insight; new therapeutic target; non-genetic; novel; release of sequestered calcium ion into cytoplasm; respiratory smooth muscle; single molecule; technological innovation

PROJECT SUMMARY: Asthma is characterized by chronic inflammation and bronchial obstruction due to human airway smooth muscle (HASM) shortening. However, the underlying basis for an enhanced shortening or the hyper-contractile state of HASM in asthma is not known. Further, our incomplete understanding of type 2 (T2) inflammation- regulated excitation-contraction (E-C) coupling in HASM shortening has hindered the development of new HASM bronchodilators with a novel mechanism of action for over 60 years. This application seeks to gain a foundational knowledge on the mechanical endotypes of HASM shortening in asthma (inflammation-dependent and -independent) and identify improved bronchodilators that are less susceptible to tolerance and less affected by immune inflammatory responses in asthma, focusing on previously unrecognized mechanisms evoked by bitter taste receptors (TAS2Rs) expressed on HASM. Our preliminary data, in pre-clinical models, support a premise that the immunologic and/or pathogenic mechanisms associated with a sustained mechanical reinforcement of HASM shortening, and the loss of β2-adrenoceptor (β2AR)-mediated bronchodilation, involve a transcriptional repressor function of the polycomb group (PcG) protein EZH2 (enhancer of zeste homolog 2). Further, our preliminary studies find a mechanistic role for microRNA-214 (miR-214) in TAS2R-evoked translational inhibition of EZH2. Based on these results, we hypothesize that TAS2Rs on HASM inhibit T2 cytokine-regulated E-C coupling in HASM shortening and the physiological loss of β2AR function in EZH2- and miR-214-dependent manners. Our goals are, first, to characterize T2- and non-T2- mediated molecular kinetics and mechanics of E-C coupling in HASM shortening and, second, determine miR- epigenetic nexus (i.e., non-genetic mechanisms) by which TAS2R activation promotes the functional efficacy of β2ARs and inhibits the mechanical endotypes of HASM shortening in asthma. Toward this end, we will leverage our unique technological innovations of single-molecule and single-cell micromechanical methods and integrative genetics and genomics approaches in clinically relevant human precision cut lung slices (hPCLS) and primary HASM cells derived from donor lungs of patients with and without severe asthma. When successful, the knowledge gained from these experimental and computational studies will: 1) shed new light on inflammation-dependent and -independent regulation of E-C coupling in HASM shortening; 2) uncover previously unidentified TAS2R paradigms to mitigate the physiological loss of β2AR function; and 3) establish new druggable targets and agents to treat β2-agonist-insenstivity in a large cohort of patients with difficult-to- control and severe asthma. This line of research is underappreciated in asthma and represents a clear shift in the asthma treatment paradigm.

项目摘要： 哮喘的特征是由于人类气道光滑而导致的慢性炎症和支气管观 肌肉（HASM）缩短。但是，增强缩短或超收缩的基础基础 哮喘中的HASM状态尚不清楚。此外，我们对2型（T2）炎症的不完全理解 - HASM缩短中的调节兴奋 - 收缩（E-C）耦合阻碍了新的发展 HASM支气管扩张剂具有新型的作用机理已有60多年的历史。本申请旨在获得 关于哮喘的HASM缩短机械内型的基础知识（炎症依赖性） 和 - 独立的），并确定不易受耐受性的支气管扩张剂的改进 受哮喘中免疫炎症反应的影响，重点是先前未知的机制 在hasm上表达的苦味受体（Tas2R）引起的诱发。我们的初步数据，在临床前模型中 支持与持续的免疫学和/或致病机制的前提 HASM缩短的机械加固，以及β2-肾上腺素受体（β2AR）介导的损失 支气管扩张，涉及Polycomb组（PCG）蛋白EZH2的转录复制函数 （Zeste同源2的增强子）。此外，我们的初步研究发现MicroRNA-214的机械作用 （miR-214）在TAS2R诱发的EZH2翻译抑制中。基于这些结果，我们假设 HASM上的TAS2R抑制hasm缩短中T2细胞因子调节的E-C耦合，并抑制了物理损失 β2AR在EZH2和MIR-214依赖性举止中的功能。我们的目标是首先要表征T2-和非T2- hasm缩短中E-C偶联的介导的分子动力学和机制，其次确定mir- TAS2R激活促进的表观遗传Nexus（即非遗传机制） β2AR并抑制哮喘中HASM缩短的机械内型。为此，我们将 利用我们独特的单分子和单细胞微力方法的技术创新 以及临床相关的人类精确切割肺切片的综合遗传学和基因组学方法 （HPCL）和主要的HASM细胞来自患有和没有严重哮喘患者的供体肺部。什么时候 成功，从这些实验和计算研究中获得的知识将：1）对 HASM缩短中E-C耦合的炎症依赖性和非依赖性调节； 2）发现 以前未识别的TAS2R范式可以减轻β2AR功能的物理损失； 3）建立 新的可药物靶标和药物治疗β2激动剂的不敏感性 控制和严重的哮喘。这种研究线在哮喘中被低估了，代表了明显的转变 哮喘治疗范例。