Rhinovirus, airway smooth muscle, and mechanisms of irreversible airflow obstruction

鼻病毒、气道平滑肌和不可逆气流阻塞机制

• 批准号: 10735460
• 负责人: Cynthia Koziol-White
• 金额: $ 50.06万
• 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: 2023-06-09 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735460
• 关键词: ADRBK1 gene; Actins; Acute; Adenosine Diphosphate; Adult; Affect; Agonist; Air; Airway Disease; Arrestin Beta 1; Asthma; Attenuated; Bronchoconstriction; Bronchodilation; Bronchodilator Agents; CXCR4 Receptors; CXCR4 gene; Calcium; Cell Line; Cell physiology; Cells; Cessation of life; Child; Clinical; Coculture Techniques; Coupled; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Development; Disease; Effector Cell; Epithelial Cells; Epithelium; Exposure to; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Health; Host Defense; Human; Immunity; Impairment; Infection; Inflammation; Inflammation Mediators; Liquid substance; Lung; MMP9 gene; Mediating; Mediator; Modeling; Molecular; Morbidity - disease rate; Muscle Contraction; Muscle relaxation phase; Myosin Light Chains; Natural regeneration; PAR-2 Receptor; Pathway interactions; Patients; Phenotype; Phosphorylation; Phosphotransferases; Pre-Clinical Model; Production; Productivity; Prostaglandins; Proteinase-Activated Receptors; Proteins; Publishing; Relaxation; Research; Rhinovirus; Rhinovirus infection; Rho-associated kinase; Role; Signal Induction; Signal Pathway; Signal Transduction; Signaling Molecule; Slice; Small Interfering RNA; Smooth Muscle; Smooth Muscle Myocytes; Symptoms; System; Therapeutic; Therapeutic Uses; Up-Regulation; VEGF165; Viral; Viral Respiratory Tract Infection; Virus; Work; airway epithelium; airway hyperresponsiveness; airway inflammation; airway obstruction; asthma exacerbation; attenuation; beta-2 Adrenergic Receptors; constriction; exposed human population; gastrointestinal epithelium; inhibitor; knock-down; loss of function; mortality; new therapeutic target; novel; phosphoric diester hydrolase; prevent; receptor; receptor coupling; receptor function; reduce symptoms; release factor 3; repaired; respiratory smooth muscle; response; therapeutic target; trefoil factor

Project Abstract: Rhinoviruses (RV) have been impugned in the development of asthma and are the leading cause of acute asthma exacerbations. RV exposure evokes inflammation of the airways, but how RV modulates human airway smooth muscle cell (HASM) function to alter bronchomotor tone is unclear. HASM is the pivotal cell modulating airway tone, shortening in response to contractile agonist stimulation through increases in intracellular calcium, through activation of Rho kinase, and through modulation of actin dynamics. We previously showed that RV exposure evokes AHR in human precision cut lung slices (hPCLS), increases [Ca2+]i in HASM, and increases agonist-induced phosphorylation of myosin light chain in HASM from human airway epithelial cell (HAEC)/HASM co-cultures stimulated with RV-C15. We also demonstrate that RVC exposure attenuates β2 agonist-induced bronchodilation in hPCLS, cAMP production in HASM, and attenuates bronchodilator-induced reversal of HASM contraction. Our preliminary data shows that TFF3, and other inflammatory mediators, are released from HAEC and hPCLS following exposure to RVC, and that TFF3 attenuates bronchodilation of human small airways and production cAMP in HASM in response to bronchodilators. Furthermore, our data demonstrates that inhibition of TFF3 and putative TFF3 receptors can reverse RV-C15-induced attenuation of bronchodilation. Therefore, we posit RV-C15 modifies signaling mechanisms modulating bronchomotor tone in HASM through a TFF3- dependent mechanism, thereby altering responsiveness of the airways to bronchodilators. We also show that signaling downstream of Gαs-coupled receptors is attenuated in HASM following exposure to RV-C15- conditioned HAEC media. We propose a central hypothesis that RVC exposure of HAEC attenuates β2 agonist-induced bronchodilation via modulation of Gαs-coupled receptor function via TFF3-dependent mechanisms. We will utilize HASM cells, air-liquid interface-differentiated HAEC, and hPCLS to examine the aims of this proposal. In Aim 1, we will determine how RV exposure modulates Gαs-coupled receptor-induced signaling to attenuate bronchodilator-induced airway/HASM relaxation and how asthma alters these mechanisms. Aim 2 will examine how RV-C15-induced TFF3 release attenuates agonist-induced bronchodilation/relaxation of HASM and how asthma alters these mechanisms. Utilizing primary HAEC/HASM co-cultures and an ex vivo system of human small airways, we will delineate signaling pathway alterations regulating bronchomotor tone that can be targeted in the treatment of RV-induced exacerbations of asthma to restore bronchodilator responsiveness.

项目摘要： 鼻病毒（RV）在哮喘的发展中受到质疑，并且是急性哮喘的主要原因。 哮喘恶化。RV暴露引起气道炎症，但RV如何调节人体气道 平滑肌细胞（HASM）改变支气管张力功能尚不清楚。HASM是细胞调节 气道张力，通过细胞内钙的增加响应于收缩激动剂刺激而缩短， 通过激活Rho激酶和通过调节肌动蛋白动力学。我们之前展示了RV 暴露引起人精确切割肺切片（hPCLS）中的AHR，增加HASM中的[Ca 2 +]i，并增加 激动剂诱导的人气道上皮细胞HASM中肌球蛋白轻链磷酸化 用RV-C15刺激的共培养物。我们还证明，RVC暴露减弱β2受体激动剂诱导的 在hPCLS中的支气管扩张，在HASM中的cAMP产生，并减弱支气管扩张剂诱导的HASM逆转 收缩。我们的初步数据表明，TFF 3和其他炎症介质，从HAEC释放， 和hPCLS，并且TFF 3减弱人小气道的支气管扩张， HASM对支气管扩张剂的反应产生cAMP。此外，我们的数据表明，抑制 TFF 3和推定的TFF 3受体可以逆转RV-C15诱导的支气管扩张衰减。所以我们 HPV-C15通过TFF 3 - 1调节HASM中支气管张力的信号传导机制。 依赖性机制，从而改变气道对支气管扩张剂的反应性。我们也证明了 暴露于RV-C15后，HASM中Gα s偶联受体下游的信号转导减弱。 条件HAEC培养基。我们提出了一个中心假设，即HAEC的RVC暴露减弱了β2 激动剂通过TFF 3依赖性调节Gα s偶联受体功能诱导支气管扩张 机制等我们将利用HASM细胞、气-液界面分化的HAEC和hPCLS来检测HASM细胞的增殖。 这一提议的目的。在目标1中，我们将确定RV暴露如何调节Gα s偶联受体诱导的 减弱支气管哮喘诱导的气道/HASM松弛的信号传导以及哮喘如何改变这些 机制等目的2将检查RV-C15诱导的TFF 3释放如何减弱激动剂诱导的TFF 3释放。 HASM的支气管扩张/松弛以及哮喘如何改变这些机制。利用初级HAEC/HASM 共培养和人类小气道的离体系统，我们将描绘信号通路的改变 调节支气管张力，可靶向治疗RV诱导的哮喘恶化， 恢复支气管扩张剂的反应性。