NHERF1 regulates MRGPRX2/MrgprB2 responses in mast cells

NHERF1 调节肥大细胞中的 MRGPRX2/MrgprB2 反应

• 批准号: 10711042
• 负责人: Hariharan Subramanian
• 金额: $ 46.07万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-12 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711042
• 关键词: Adaptor Signaling Protein; Affect; Agonist; Allergic; Allergic Disease; Allergic Reaction; Anaphylaxis; Asthma; Attenuated; Biological Assay; CD34 gene; CISH gene; Cell Compartmentation; Cell physiology; Cells; Cellular biology; Chronic; Clinical; Data; Development; Disease; Edema; Emergency department visit; Engraftment; Expenditure; Female; Future; G-Protein-Coupled Receptors; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Healthcare; Human; IL3 Gene; IgE Receptors; Immune; In Vitro; Inflammation; Inflammatory; Intervention; Ligands; Mediating; MicroRNAs; Molecular; Mouse Strains; Mus; Orthologous Gene; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Phosphorylation; Play; Proteins; Proteomics; Reaction; Regulation; Research; Role; Rosacea; Signal Pathway; Signal Transduction; Signaling Molecule; Signaling Protein; Site; Testing; Time; Tissues; Umbilical Cord Blood; United States Food and Drug Administration; Urticaria; allergic response; cathelicidin; cathelicidin antimicrobial peptide; chronic inflammatory disease; clinical application; cytokine; drug development; experimental study; humanized mouse; in vivo; in vivo Model; inositol-1,4,5-trisphosphate 5-phosphatase; insight; mast cell; mastocytosis; mimetics; mortality; mouse model; mutant; novel; novel therapeutics; overexpression; pharmacologic; phosphoproteomics; receptor; response; sodium-hydrogen exchanger regulatory factor; stem cells; transcriptomics

PROJECT SUMMARY Mast cells are tissue resident innate immune cells that have been best characterized for their role in mediating allergic diseases. A new development in mast cell research has been the identification and characterization of a novel G-protein coupled receptor, Mas-Related G-Protein-coupled Receptor X2 (MRGPRX2). This receptor promotes pseudoallergic reaction to U.S. Food and Drug Administration (FDA)-approved drugs and chronic inflammation in diseases such as asthma, rosacea and hives (urticaria). While the role of MRGPRX2 in promoting allergic reactions has been thoroughly investigated, the molecular mechanisms utilized by this receptor is poorly understood. Our data suggests that Na+/H+ exchanger regulatory factor (NHERF)1, an adaptor protein, regulates MRGPRX2- and MrgprB2- (the mouse ortholog of the human receptor) induced responses in mast cells. Specifically, reducing the expression of NHERF1 in mast cells results in decreased intracellular Ca2+ mobilization and degranulation via MRGPRX2/MrgprB2 in vitro. Consistent with this data, NHERF1 also promoted anaphylaxis in MrgprB2-dependent mouse models of paw edema and passive systemic anaphylaxis in vivo. This proposal builds up on these observations; the central hypothesis is that NHERF1 is a critical regulator of MRGPRX2 responses in mast cells. Using a novel mouse strain that we have recently generated, Cpa3- Cre+:NHERF1fl/fl, primary human CD34+ stem cell-derived mast cells and humanized mice, we will determine the role of mast cell-specific expression of NHERF1 in regulating mouse MrgprB2 and human MRGPRX2 receptors in vitro and in vivo (Aim 1). In Aim 2, we will explore the mechanisms through which NHERF1 modulates mast cell response. Specifically, we will determine whether phosphorylation of NHERF1 is important for NHERF1 regulation of MRGPRX2/MrgprB2. We will identify the components of the NHERF1-interactome and signaling proteins and pathways through which NHERF1 functions in mast cells by using transcriptomic and proteomic approaches. Additionally, we will delineate the role of NHERF1-microRNA (miR)155-suppressor of cytokine signaling (SOCS1)-Src homology 2 (SH2) domain containing inositol polyphosphate 5-phosphatase 1 (SHIP-1) axis in promoting mast cell activation. Given the critical role of mast cell-MRGPRX2 in causing pseudoallergic reactions and chronic inflammation in allergic diseases, elucidation of the mechanisms by which NHERF1 contributes to MRGPRX2-mediated allergic response is of significant scientific and clinical importance. We firmly believe that successful completion of the proposed studies will likely lead to the identification of potential targets of the MRGPRX2 pathway that can provide insights into the future development of drugs for not only pseudoallergic reactions but also other mast cell-mediated inflammatory diseases.

项目总结 肥大细胞是一种驻留在组织中的先天免疫细胞，因其在 过敏性疾病。肥大细胞研究的一个新进展是鉴定和鉴定了一种 新的G蛋白偶联受体，Mas相关G蛋白偶联受体X2(MRGPRX2)。这种受体 促进对美国食品和药物管理局(FDA)批准的药物和慢性药物的假性过敏反应 哮喘、酒渣鼻和麻疹等疾病的炎症。而MRGPRX2在促进 过敏反应已被深入研究，但该受体利用的分子机制较差。 明白了。我们的数据表明，Na+/H+交换调节因子(NHERF)1，一种适配器蛋白，调节 MRGPRX2-和MRGPRB2-(人类受体的小鼠同源基因)在肥大细胞中诱导反应。 具体地说，减少肥大细胞中NHERF1的表达会导致细胞内钙离子动员减少 体外用MRGPRX2/MRgprB2脱颗粒。与此数据一致的是，NHERF1还推动了 在活体内对mrgprB2依赖的小鼠足爪肿胀和被动全身过敏反应模型的过敏反应。这 建议建立在这些观察的基础上；中心假设是NHERF1是一个关键的调节因子 肥大细胞中的MRGPRX2反应。使用我们最近产生的一种新的小鼠品系，Cpa3- Cre+：NHERF1fl/fl，原代人CD34+干细胞来源的肥大细胞和人源化小鼠，我们将确定 肥大细胞特异性表达NHERF1在调节小鼠MRGPRB2和人MRGPRX2受体中的作用 体外和体内实验(目标1)。在目标2中，我们将探索NHERF1调节MAST的机制 细胞反应。具体地说，我们将确定NHERF1的磷酸化是否对NHERF1重要 MRGPRX2/MRgprB2的调控。我们将确定NHERF1的组件--互动组和信号 利用转录和蛋白质组学研究肥大细胞中NHERF1功能的蛋白质和途径 接近了。此外，我们还将描述NHERF1-microRNA(MiR)155-细胞因子抑制因子的作用 信号转导(SOCS1)-含肌醇多聚磷酸5-磷酸酶1(SHIP-1)的Src同源2(SH2)结构域 轴在促进肥大细胞激活中的作用。鉴于肥大细胞MRGPRX2在引起假性变态反应中的关键作用 过敏性疾病中的反应和慢性炎症，阐明NHERF1的机制 参与MRGPRX2介导的过敏反应具有重要的科学和临床意义。我们坚定地 我相信成功完成拟议的研究可能会导致确定潜在的目标 对MRGPRX2途径的研究，不仅可以为药物的未来发展提供见解 假性过敏反应，也包括其他肥大细胞介导的炎症性疾病。