RBM3 regulation of type 2 innate lymphoid cells in allergic inflammation

RBM3 对过敏性炎症中 2 型先天淋巴细胞的调节

• 批准号: 8799448
• 负责人: TAYLOR A DOHERTY
• 金额: $ 47.54万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-11-10 至 2019-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8799448
• 关键词: Adoptive Transfer; Allergens; Allergic; Allergic Disease; Allergic inflammation; Amino Acid Motifs; Animal Model; Asthma; B-Lymphocytes; Biological Assay; Biology; Blood; Cells; Data; Elements; Epithelial; Exclusion; Extrinsic asthma; Future; Genetic; Goals; Human; Immune; In Vitro; Infiltration; Interleukin-13; Interleukin-5; Knockout Mice; Leukocytes; Lung; Lung Inflammation; Lymphocyte; Lymphoid Cell; Mediating; Messenger RNA; Modeling; Molecular; Monitor; Mucous body substance; Mus; Nasal Polyps; Northern Blotting; Pathogenesis; Pathway interactions; Phenotype; Population; Production; RNA Binding; RNA-Binding Proteins; Reagent; Regulation; Reporter; Research; Role; Small Interfering RNA; T-Lymphocyte; TSLP gene; Testing; Time; Transcript; airway hyperresponsiveness; airway inflammation; antigen challenge; base; cell type; cytokine; eosinophil; in vivo; knock-down; mRNA Stability; new therapeutic target; novel; public health relevance; response; therapeutic target

DESCRIPTION (provided by applicant): The goal of the proposed research is to identify novel mechanisms that regulate type 2 innate lymphoid cells (ILC2). ILC2 are a recently discovered population of lineage-negative lymphocytes that produce Th2 cytokines in response to epithelial cytokines IL-33 and TSLP. ILC2 contribute to an asthma phenotype in animal models and are present in human lungs. Specifically, we hypothesize that the RNA-binding protein RBM3 controls human and mouse ILC2 Th2 cytokine production and innate type 2 lung responses through Th2 cytokine mRNA stabilization. We will administer allergen, IL-33, and TSLP challenges to the airways of WT and RBM3 knockout mice and assess lung ILC2 Th2 cytokine production and proliferation, as well as levels of BAL eosinophils, epithelial mucus production, peribronchial infiltration and airway hyperresponsiveness. We will also perform ILC2 adoptive transfer studies and generate RBM3 deficient IL-13 reporter mice to define the role of RBM3 in ILC2 during allergen, IL-33, and TSLP-induced airway inflammation. We will perform in vitro studies with purified ILC2 from WT and RBM3 knockout mice as well as human RMB3 siRNA knockdown ILC2 stimulated with IL-33 and TSLP and assess levels of ILC2 Th2 cytokine production and proliferation. Further, we will perform mRNA stability assays to determine whether RBM3 controls ILC2 function by stabilizing Th2 cytokine mRNAs. We expect to find that RMB3 promotes ILC2 Th2 cytokine mRNA stability and controls ILC2 Th2 cytokine production in vitro and in vivo. The novelty of the proposed studies includes use of novel reagents (RBM3-/-, RBM3-/-RAG2-/-, and RBM3-/-/YFP-IL-13 reporter mice) and uncovering a novel pathway (RNA binding protein) regulating a recently discovered cell type (ILC2) that contributes to allergic inflammation. Importantly, the discovery of molecules that regulate ILC2 function may reveal novel therapeutic targets to reduce ILC2 activation in asthma and allergic disease.

描述（由申请人提供）：拟议研究的目标是确定调节2型先天淋巴样细胞（ILC2）的新机制。ILC2是最近发现的一种谱系阴性淋巴细胞，其产生Th2细胞因子以响应上皮细胞因子IL-33和TSLP。在动物模型中，ILC2有助于哮喘表型，并存在于人肺中。具体来说，我们假设rna结合蛋白RBM3通过稳定Th2细胞因子mRNA来控制人和小鼠ILC2 Th2细胞因子的产生和先天2型肺反应。我们将对WT和RBM3基因敲除小鼠的气道给予过敏原、IL-33和TSLP挑战，并评估肺ILC2 Th2细胞因子的产生和增殖，以及BAL嗜酸性粒细胞、上皮粘液产生、支气管周围浸润和气道高反应性的水平。我们还将进行ILC2过继性转移研究，并生成RBM3缺陷的IL-13报告小鼠，以确定在过敏原、IL-33和tslp诱导的气道炎症期间，RBM3在ILC2中的作用。我们将对WT和RBM3敲除小鼠的纯化ILC2以及人RMB3 siRNA敲除ILC2进行体外研究，并通过IL-33和TSLP刺激ILC2，评估ILC2 Th2细胞因子的产生和增殖水平。此外，我们将进行mRNA稳定性分析，以确定RBM3是否通过稳定Th2细胞因子mRNA来控制ILC2功能。我们希望在体外和体内发现RMB3促进ILC2 Th2细胞因子mRNA的稳定性并控制ILC2 Th2细胞因子的产生。这些研究的新颖之处包括使用新试剂（RBM3-/-、RBM3-/- rag2 -/-和RBM3-/-/YFP-IL-13报告小鼠）和发现一种新途径（RNA结合蛋白）调节最近发现的一种有助于过敏性炎症的细胞类型（ILC2）。重要的是，发现调节ILC2功能的分子可能揭示新的治疗靶点，以减少哮喘和过敏性疾病中ILC2的激活。