Oxysterol Regulation of Mast Cell Biology

肥大细胞生物学的氧甾醇调节

• 批准号: 10237850
• 负责人: Daniel F Dwyer
• 金额: $ 10.8万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-13 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10237850
• 关键词: 25-hydroxycholesterol; 7alpha hydroxylase; Address; Airway Disease; Allergens; Allergic; Asthma; Atopic Dermatitis; Automobile Driving; Biological Models; Cell Compartmentation; Cell Count; Cell Culture System; Cell Degranulation; Cell Lineage; Cell Survival; Cells; Cellular biology; Characteristics; Chemotaxis; Cholesterol; Collaborations; Complement 3d Receptors; Contact Dermatitis; Data; Data Set; Disease; Doctor of Philosophy; Ear; Effector Cell; Enzymes; Eosinophilic Esophagitis; Epithelial Cells; Future; G-Protein-Coupled Receptors; GTP-Binding Protein alpha Subunits, Gs; Gene Expression; Generations; Genetic Transcription; Histamine; Human; Human Herpesvirus 4; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Inhalation; Interleukin-13; Laboratories; Ligands; Ligation; Lung; Lung Inflammation; Manuscripts; Mediating; Mixed Function Oxygenases; Modeling; Mouse Strains; Mucous Membrane; Mus; Myelogenous; Nasal Polyps; Orphan; Oxides; Passive Cutaneous Anaphylaxis; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Peptide Hydrolases; Play; Polyps; Production; Proto-Oncogene Protein c-kit; Pulmonary Inflammation; Regulation; Reporting; Research; Role; Sampling; Swelling; Testing; Therapeutic; Tissues; Transcript; United States National Institutes of Health; Up-Regulation; airway epithelium; airway hyperresponsiveness; asthmatic; asthmatic airway smooth muscle; career; cell motility; chronic rhinosinusitis; cytokine; human tissue; imprint; in vitro Assay; in vivo; intradermal injection; lipid mediator; mast cell; mouse model; mucosal site; multiple datasets; novel; oxysterol 7-alpha-hydroxylase; oxysterol binding protein; receptor; recruit; respiratory; single-cell RNA sequencing; stem cells; targeted treatment; tool; virtual

PROJECT SUMMARY/ABSTRACT This proposal details a two-year plan to allow the candidate, Daniel Dwyer, PhD, to transition to a stable independent research career. The focus of this study is on characterizing a novel regulatory axis capable of mediating the recruitment and activation of mast cells (MCs) during type 2 inflammation (T2I). MCs are potent effector cells that play key pathogenic roles in type 2 inflammatory diseases through the generation and release of a broad range of inflammatory mediators, including histamine, proteases, lipid mediators, and cytokines. The specific mechanisms underlying the expansion of MC during type 2 inflammation or the persistent activation of these cells associated with these diseases are unclear. This proposal identifies novel expression of a G protein-coupled receptor (GPCR) in both human and mouse MCs. Ligation of this receptor elicits mediating MC migration in vitro and additionally induces MC activation both in vivo and in vitro. Further, this proposal further identifies upregulation of a necessary enzyme for synthesis of the ligand across a spectrum of human T2I disease and finds that murine MC progenitor recruitment during allergic pulmonary inflammation is virtually absent in mice lacking the enzyme. Aim 1 of this proposal will utilize two murine models of allergic lung inflammation to assess which cells express and upregulate the enzyme in the context of T2I, followed by in vitro approaches to determine the mechanism driving this upregulation. Aim 2 of this proposal details the generation of a novel mouse strain in which the GPCR is specifically deleted in MC. After an initial confirmation of specific deletion and characterization of the MC compartment across tissues, the specific role of this GPCR in regulating constitutive MC activation and MC progenitor recruitment will be assessed in two inflammatory models.

项目总结/摘要 这份提案详细说明了一个为期两年的计划，让候选人，丹尼尔德怀尔，博士，过渡到一个稳定的 独立的研究生涯。这项研究的重点是表征一种新的调节轴， 在2型炎症（T2 I）期间介导肥大细胞（MC）的募集和活化。MC很强大 在2型炎性疾病中发挥关键致病作用的效应细胞， 释放广泛的炎性介质，包括组胺、蛋白酶、脂质介质和 细胞因子在2型炎症过程中MC扩张的具体机制或 与这些疾病相关的这些细胞的持续活化尚不清楚。该提案确定了新颖的 G蛋白偶联受体（GPCR）在人类和小鼠MC中的表达。该受体的连接 在体外介导MC迁移，并且另外在体内和体外诱导MC活化。此外，本发明还 该建议进一步鉴定了在细胞内合成配体所必需的酶的上调， 人类T2 I疾病谱，并发现小鼠MC祖细胞招募在过敏性肺 缺乏这种酶的小鼠几乎没有炎症。本提案的目标1将利用两个小鼠 过敏性肺部炎症模型，以评估哪些细胞表达和上调酶的背景下， T2 I，然后通过体外方法来确定驱动这种上调的机制。目标2 该提案详细描述了一种新的小鼠品系的产生，其中GPCR在MC中特异性缺失。后 初步确认特异性缺失和跨组织MC区室的表征， 该GPCR在调节组成性MC活化和MC祖细胞募集中的特定作用将被 在两种炎症模型中评估。