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.

项目概要/摘要 该提案详细介绍了一项两年计划，旨在让候选人 Daniel Dwyer 博士过渡到稳定的岗位 独立的研究生涯。这项研究的重点是描述一个新的监管轴，该轴能够 在 2 型炎症 (T2I) 期间介导肥大细胞 (MC) 的募集和激活。 MC很给力 效应细胞通过产生和形成在 2 型炎症性疾病中发挥关键致病作用 释放多种炎症介质，包括组胺、蛋白酶、脂质介质和 细胞因子。 2 型炎症期间 MC 扩张的具体机制或 这些细胞的持续激活与这些疾病相关尚不清楚。该提案确定了新颖的 G 蛋白偶联受体 (GPCR) 在人和小鼠 MC 中的表达。该受体的连接 在体外引发介导 MC 迁移，并在体内和体外诱导 MC 激活。更远， 该提议进一步确定了配体合成所必需的酶的上调 人类 T2I 疾病谱并发现小鼠 MC 祖细胞在过敏性肺病期间招募 缺乏这种酶的小鼠实际上不存在炎症。该提案的目标 1 将利用两只小鼠 过敏性肺部炎症模型，以评估哪些细胞在以下情况下表达和上调该酶 T2I，然后通过体外方法来确定驱动这种上调的机制。这个目标2 该提案详细介绍了一种新型小鼠品系的产生，其中 GPCR 在 MC 中被特异性删除。后 跨组织的 MC 区室的特异性删除和表征的初步确认， 该 GPCR 在调节组成型 MC 激活和 MC 祖细胞招募中的具体作用将是 在两种炎症模型中进行评估。