Role of beta-arrestin-2 on IgE-mediated cofilin dephosphorylation and mast cell activation

beta-arrestin-2 对 IgE 介导的丝切蛋白去磷酸化和肥大细胞激活的作用

基本信息

批准号：
9114460
负责人：
Hydar Ali
金额：
$ 24万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9114460
关键词：
ADRBK1 gene AIDS/HIV problem ARRB1 gene ARRB2 Actins Adrenal Cortex Hormones Affinity Agonist Allergens Antibodies Antigens Area Arr2 Arrestins Asthma Attenuated Binding Sites Bone Marrow Breeding Carboxypeptidase Cells Chemotaxis Chronic Complement Complement 3a Complex Data Development Disease Extrinsic asthma Family G Protein-Coupled Receptor Signaling G protein coupled receptor kinase G-Protein-Coupled Receptors GTP-Binding Proteins Generations Goals Health Histamine Release Human IgE IgE Receptors Immune response In Vitro Inflammation Mediators Inflammatory Laboratories Leukotriene Antagonists Lipids LoxP-flanked allele Lung Lung diseases MAPK Signaling Pathway Pathway Mediating Medical Morbidity - disease rate Mus Pathogenesis Patients Phosphorylation Phosphotransferases Play Plus End of the Actin Filament Protein Dephosphorylation Protein Kinase Proteins Receptor Signaling Regulation Reporting Retroviridae Role Scaffolding Protein Signal Pathway Signal Transduction Signaling Protein Tertiary Protein Structure Testing Therapeutic Tuberculosis United States antimicrobial peptide base beta-arrestin cofilin cysteinyl-leukotriene cytokine depolymerization desensitization economic cost in vivo lipid mediator mast cell mastocytosis montelukast novel novel strategies novel therapeutic intervention novel therapeutics omalizumab overexpression protein activation receptor response scaffold

项目摘要

DESCRIPTION (provided by applicant): Asthma is one of the most common respiratory diseases world-wide; 300 million people are currently receiving treatment and global economic cost exceeds those of tuberculosis and HIV/AIDS combined. Thus, asthma remains an area of considerable unmet medical need. Allergic asthma is caused by an overzealous Th2 immune response to allergens in which immunoglobulin E (IgE) and mast cells play critical roles. Thus, aggregation of high affinity IgE receptor (FcεRI) by allergen on mast cells results in histamine release and the generation of lipids and cytokines, which are responsible for the manifestations of asthma. The focus of our laboratory has been to study the regulation of G protein coupled receptor (GPCR) signaling in mast cells. Functions of GPCRs are regulated via their phosphorylation by a family of protein kinases known as GPCR kinases (GRKs) and the recruitment of the scaffolding molecule ß-arrestin (ß-arr). The receptor/ß-arr complex serves to attenuate G protein activation leading to receptor desensitization. ß-arr also promotes downstream MAPK signaling pathways independent of G protein activation. To the best of our knowledge, the role of ß-arr on FcεRI signaling in mast cells has never been reported. Based on our unexpected preliminary data, we hypothesize that ß-arr2 serves as a scaffolding protein to promote FcεRI- mediated cofilin dephosphorylation in mast cells. We further hypothesize that this ß-arr2-mediated cofilin dephosphorylation contributes to mast cell recruitment in the lung to promote the development chronic asthma. In aim 1, retrovirus will be used to transduce different domains of ß-arr2 into ß-arr2-/- mast cells and their ability to promote FcεRI-mediated cofilin dephosphorylation and mast cell chemotaxis will be determined. We will then overexpress different sub-domains of ß-arr2 into mast cells endogenously expressing ß-arr2 and their ability to interfere with FcεRI-mediated cofilin dephosphorylation and mast cell chemotaxis will be determined. In aim 2, we will delete ß-arr2 in mast cells by breeding ß-arr2-floxed mice and carboxypeptidase-3 (Cpa-3) cre mice. We will also overexpress ß-arr2 in mast cells in vivo by engrafting ß-arr2-transduced BMMCs into mast cell-deficient Wsh/Wsh mice. These complementary approaches will be used to test the hypothesis that ß-arr2 expressed in mast cells contributes to their recruitment in the lung and promotes the development of chronic asthma. Finally, we will express cofilin and peptide domains of ß-arr2 in mast cells in vivo. These strategies will be used to determine if mast cell-specific expression of cofilin and ß-arr2 domains modulate chronic asthma. This study will likely generate new information regarding the roles of ß-arr2 and cofilin in mast cell chemotaxis in vitro and in vivo and may provide a better rationale for the development of novel therapeutics for chronic asthma.

描述（由申请人提供）：哮喘是全世界最常见的呼吸道疾病之一；目前有 3 亿人正在接受治疗，全球经济成本超过了结核病和艾滋病毒/艾滋病的总和。因此，哮喘仍然是一个领域。大量未满足的医疗需求是由对过敏原过度热衷的 Th2 免疫反应引起的，其中免疫球蛋白 E (IgE) 和肥大细胞发挥着关键作用，因此，高亲和力的聚集。肥大细胞上过敏原的 IgE 受体 (FcεRI) 会导致组胺释放以及脂质和细胞因子的产生，从而导致哮喘的表现。我们实验室的重点是研究 G 蛋白偶联受体 (GPCR) 的调节。 GPCR 的功能通过称为 GPCR 激酶 (GRK) 的蛋白激酶家族的磷酸化和支架分子 ß-arrestin 的募集来调节。 (ß-arr)。受体/ß-arr 复合物的作用是据我们所知，β-arr 还可以促进下游 MAPK 信号转导通路，从而导致肥大细胞中 FcεRI 信号转导的作用。我们意想不到的初步数据表明，β-arr2 作为支架蛋白可促进肥大细胞中 FcεRI 介导的肌动蛋白丝切蛋白去磷酸化。 ß-arr2 介导的丝切蛋白去磷酸化有助于肥大细胞在肺中募集在目标 1 中，逆转录病毒将用于将 ß-arr2 的不同结构域转导至 ß-arr2-/- 肥大细胞，并确定其促进 FcεRI 介导的丝切蛋白去磷酸化和肥大细胞趋化性的能力。然后将 ß-arr2 的不同子结构域过表达到内源表达 ß-arr2 的肥大细胞中，并具有干扰 FcεRI 介导的能力在目标 2 中，我们将通过培育 ß-arr2-floxed 小鼠和羧肽酶-3 (Cpa-3) cre 小鼠来删除肥大细胞中的 ß-arr2。通过将 ß-arr2 转导的 BMMC 移植到肥大细胞缺陷的 Wsh/Wsh 小鼠体内，在体内肥大细胞中产生肥大细胞。我们将使用补充方法来检验肥大细胞中表达的 ß-arr2 有助于肥大细胞在肺部募集并促进慢性哮喘发展的假设。最后，我们将在体内肥大细胞中表达 ß-arr2 的丝动蛋白和肽结构域。这些策略将用于确定肥大细胞特异性表达 cofilin 和 ß-arr2 结构域是否调节慢性哮喘。这项研究可能会产生有关 ß-arr2 和 cofilin 在哮喘中的作用的新信息。肥大细胞在体外和体内的趋化性可能为开发慢性哮喘的新疗法提供更好的理由。