Regulation of Normal and Asthmatic Lung Function by G-Protein-Coupled Receptors

G 蛋白偶联受体对正常和哮喘肺功能的调节

• 批准号: 10927794
• 负责人: Kirk m Druey
• 金额: $ 63.4万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10927794
• 关键词: Actins; Adenosine; Adrenal Cortex Hormones; Adrenergic beta-Agonists; Affect; Affinity; Agonist; Albuterol; Allergens; Antifungal Agents; Area; Aspergillus fumigatus; Asthma; Autopsy; Binding; Bradykinin; Bronchoconstriction; Bronchodilator Agents; Calcium; Cells; Cessation of life; Collaborations; Coupled; Cyclic AMP; Disease susceptibility; Drug Targeting; Endothelin-1; Extracellular Matrix; Extrinsic asthma; G Protein-Coupled Receptor Signaling; G alpha q Protein; G-Protein-Coupled Receptors; GTP-Binding Protein Regulators; GTP-Binding Protein alpha Subunits; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Gene Deletion; Goals; Grant; Guanosine Triphosphate Phosphohydrolases; Hepatitis C; Hepatitis C Therapy; Heterotrimeric GTP-Binding Proteins; Histamine; Human; Hypersensitivity; IgE; In Vitro; Infection; Inflammation Mediators; Inflammatory Response; Inhibitory G-Protein Gi; Lead; Leukotriene Antagonists; Leukotriene D4; Life; Ligands; Link; Lung; Mediating; Modeling; Molds; Monoclonal Antibodies; Mus; Muscle Contraction; Muscle Fibers; Muscle function; Muscle relaxation phase; Myosin ATPase; Natural Substance; PAR-2 Receptor; Pathologic; Peptide Hydrolases; Persons; Physiological; Proliferating; Pulmonary Inflammation; RGS Domain; RGS Proteins; Regulation; Relaxation; Role; Serine Protease; Signal Induction; Signal Pathway; Signal Transduction; Slice; Smooth Muscle; Smooth Muscle Myocytes; Specimen; Therapeutic; Therapeutic Uses; Thrombin; Tissue Banks; Tissues; aggressive therapy; airway hyperresponsiveness; airway obstruction; allergic airway inflammation; asthmatic; asthmatic airway; cysteinyl-leukotriene; efficacy study; eosinophil; eosinophilic inflammation; experience; fungus; granule cell; inhibitor; insight; interest; mast cell; mouse model; novel; novel therapeutics; omalizumab; pulmonary function; receptor; respiratory; respiratory smooth muscle; success

Asthma, a pathological condition of reversible airway obstruction, is comprised of both inflammation of the lung and hyper-contractility of the bronchial smooth muscle. The major naturally occurring substances that induce bronchial smooth muscle contraction are ligands of G-protein-coupled receptors (GPCRs), such as allergen proteases, thrombin, and those contained in allergen-IgE activated mast cell granules (e.g. histamine, cysteinyl leukotrienes (LTD4), endothelin 1, adenosine, and bradykinin). In general, these agonists induce activation of the heterotrimeric G protein G-alpha q, which increases the concentration of intracellular calcium in smooth muscle cells, promoting actin-myosin interactions and muscle fiber shortening. In contrast, ligands acting on G-alpha-s-coupled receptors, such as albuterol, increase intracellular levels of cyclic AMP (cAMP), facilitating ASM relaxation. Although eosinophilic inflammation typifies allergic asthma, it is not a prerequisite for airway hyper-responsiveness (AHR), suggesting that underlying abnormalities in structural cells including ASM contribute to the asthmatic diathesis. Dysregulation of procontractile, GPCR signaling in ASM could mediate enhanced contractility. 10-15% of people with asthma experience severe, life threatening attacks and even death despite aggressive treatment with bronchodilators and corticosteroids. Nearly half of these (10-20 million) are sensitized (i.e. have IgE-mediated allergy) to filamentous fungi (e.g. Aspergillus fumigatus, Af), which has been designated "severe asthma with fungal sensitization" (SAFS). Current therapies for SAFS including antifungals or omalizumab monoclonal antibody (mAb) targeting IgE have not achieved uniform success. Utilizing a model of allergic airway inflammation in mice induced by respiratory Af exposure, we have 2 overarching aims for this project: 1) identify derangements in ASM contraction signaling downstream of inflammatory mediators; 2) examine the functions of allergen protease activity in AHR, particularly in relation to allergen-ASM interactions. Protease activity is a common and important feature of allergens capable of inducing asthma, most notably from ubiquitous fungi such as Af. Whether any allergens affect ASM contraction directly has never been explored. Previously we found a causal link between fungi and asthma occurring independently of allergenicity; in other words, host inflammatory response to the allergen. The secreted Af protease Alkaline protease 1 (Alp1) was detected in the airways of asthmatic subjects but not controls. In mouse and in vitro studies, we found that Alp1 directly promoted AHR by degrading extracellular matrix (ECM) components, leading to dysregulation of ASM contraction. Subsequently, we demonstrated that Alp1 induces AHR in mice devoid of eosinophils or the protease activated receptor 2 (PAR2) and that Alp1 directly increased contractile force of human ASM cells in vitro. In Fiscal Year (FY) 23, identified an inhibitor of Alp1 protease activity in a compound screen done in collaboration with Dr. Krishnan through an R21 grant. A critical therapeutic gap is to identify moieties with specific affinity for fungal Alp1 but without activity on endogenous serine proteases present in the human airway. The lead compound, which is used therapeutically for hepatitis C (HCV) infections, inhibits the effect of Alp on ASM contraction in vitro and in human airways of murine and human precision-cut lung slices (PCLS) treated with Alp1. We are currently studying the efficacy of the compound in mouse models of fungal asthma. We expect these studies to identify a novel therapeutic application for an HCV inhibitor for fungal asthma and provide novel insight into both ASM-intrinsic and allergen-protease-dependent mechanisms of bronchoconstriction. The second area of emphasis for this project is the study of Regulators of G protein signaling (RGS) proteins in the lung. RGSs bind to the G protein alpha subunits Gi and Gq (but not Gs) through a conserved RGS domain and inactivates them by catalyzing their intrinsic GTPase activity and by blocking downstream effector interactions. Although they are generally considered to act as negative regulators of GPCR signaling pathways, the physiological function of RGS proteins in the lung is mostly unknown. We identified expression of several RGS proteins (RGS4, RGS5) in bronchial smooth muscle of humans and mice.

哮喘是一种可逆性气道阻塞的病理状态，由肺部炎症和支气管平滑肌过度收缩组成。诱导支气管平滑肌收缩的主要天然物质是g蛋白偶联受体（gpcr）的配体，如过敏原蛋白酶、凝血酶和过敏原ige激活的肥大细胞颗粒中含有的配体（如组胺、半胱氨酸白三烯（LTD4）、内皮素1、腺苷和缓激肽）。一般来说，这些激动剂诱导异三聚体G蛋白G- α q的激活，从而增加平滑肌细胞内钙的浓度，促进肌动蛋白-肌球蛋白相互作用和肌纤维缩短。相反，作用于g - α -s偶联受体的配体，如沙丁胺醇，增加细胞内环AMP （cAMP）水平，促进ASM松弛。虽然嗜酸性粒细胞炎症是过敏性哮喘的典型特征，但它并不是气道高反应性（AHR）的先决条件，这表明包括ASM在内的结构细胞的潜在异常有助于哮喘素质。ASM中促收缩性、GPCR信号的失调可能介导收缩性增强。

项目成果

Regulation of G-protein-coupled signaling pathways in allergic inflammation.

• DOI: 10.1007/s12026-008-8050-0
• 发表时间: 2009
• 期刊: IMMUNOLOGIC RESEARCH
• 影响因子: 4.4
• 作者: Druey, Kirk M.

Alternaria alternata Accelerates Loss of Alveolar Macrophages and Promotes Lethal Influenza A Infection.

• DOI: 10.3390/v12090946
• 发表时间: 2020-08-27
• 期刊: Viruses
• 作者: Percopo CM;Ma M;Mai E;Redes JL;Kraemer LS;Minai M;Moore IN;Druey KM;Rosenberg HF
• 通讯作者: Rosenberg HF

Persistent Airway Hyperresponsiveness Following Recovery from Infection with Pneumonia Virus of Mice.

• DOI: 10.3390/v13050728
• 发表时间: 2021-04-22
• 期刊: Viruses
• 作者: Limkar AR;Percopo CM;Redes JL;Druey KM;Rosenberg HF
• 通讯作者: Rosenberg HF

Alternaria alternata challenge at the nasal mucosa results in eosinophilic inflammation and increased susceptibility to influenza virus infection.

• DOI: 10.1111/cea.13123
• 发表时间: 2018-06
• 期刊: Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology
• 作者: Ma M;Redes JL;Percopo CM;Druey KM;Rosenberg HF
• 通讯作者: Rosenberg HF

Modeling asthma: Pitfalls, promises, and the road ahead.

• DOI: 10.1002/jlb.3mr1117-436r
• 发表时间: 2018-07
• 期刊: Journal of leukocyte biology
• 影响因子: 5.5
• 作者: Rosenberg HF;Druey KM
• 通讯作者: Druey KM