RBM3 regulation of type 2 innate lymphoid cells in allergic inflammation

描述（由申请人提供）：拟议研究的目标是确定调节2型先天淋巴细胞（ILC 2）的新机制。ILC 2是最近发现的谱系阴性淋巴细胞群体，其响应于上皮细胞因子IL-33和TSLP而产生Th 2细胞因子。ILC 2有助于动物模型中的哮喘表型，并且存在于人肺中。具体而言，我们假设RNA结合蛋白RBM 3通过Th 2细胞因子mRNA稳定化控制人和小鼠ILC 2 Th 2细胞因子的产生和先天性2型肺应答。我们将对WT和RBM 3基因敲除小鼠的气道给予过敏原、IL-33和TSLP激发，并评估肺ILC 2 Th 2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ILC 2过继转移研究，并产生RBM 3缺陷型IL-13报告小鼠，以确定RBM 3在ILC 2中在过敏原、IL-33和TSLP诱导的气道炎症中的作用。我们将用来自WT和RBM 3敲除小鼠的纯化ILC 2以及用IL-33和TSLP刺激的人RMB 3 siRNA敲低ILC 2进行体外研究，并评估ILC 2 Th 2细胞因子产生和增殖的水平。此外，我们将进行mRNA稳定性测定以确定RBM 3是否通过稳定Th 2细胞因子mRNA来控制ILC 2功能。我们期望在体外和体内发现RMB 3促进ILC 2 Th 2细胞因子mRNA的稳定性并控制ILC 2 Th 2细胞因子的产生。拟议研究的新奇包括使用新型试剂（RBM 3-/-、RBM 3-/-RAG 2-/-和RBM 3-/-/YFP-IL-13报告小鼠），并揭示了一种调节最近发现的细胞类型（ILC 2）的新型途径（RNA结合蛋白），该细胞类型有助于过敏性炎症。重要的是，调控ILC 2功能的分子的发现可能揭示新的治疗靶点，以减少哮喘和过敏性疾病中ILC 2的激活。

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.