MicroRNA directed pathway discovery in allergy and asthma

MicroRNA 指导过敏和哮喘通路的发现

• 批准号: 10206229
• 负责人: Karl Mark Ansel
• 金额: $ 40.38万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10206229
• 关键词: ARID1A gene; Acute; Airway Disease; Allergens; Allergic; Antibodies; Asthma; B-Lymphocytes; Basophils; Binding; Binding Sites; Biochemical; Bioinformatics; Biological; Biological Process; Biology; CRISPR/Cas technology; Catalogs; Cell Differentiation process; Cells; Chronic; Communication; Computer Analysis; Data; Development; Disease; Epithelial Cells; Extrinsic asthma; FOXP1 gene; Family; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Helper-Inducer T-Lymphocyte; Histamine Release; Human; Hypersensitivity; IgE; Immunoglobulin Class Switching; Immunoglobulin Switch Recombination; Immunoglobulin-Secreting Cells; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Knockout Mice; Knowledge; Lead; Link; Maps; Mediating; Messenger RNA; MicroRNAs; Molecular; Mus; Mutant Strains Mice; Pathogenesis; Pathway interactions; Patients; Physiological; Plasma Cells; Prevention approach; Production; Property; Protocols documentation; Regulator Genes; Research; Selection Bias; Symptoms; System; T cell differentiation; T cell response; Techniques; Therapeutic antibodies; Tissues; Translations; Tumor-infiltrating immune cells; airborne allergen; airway epithelium; arm; asthma model; base; cell behavior; comparative; conditional knockout; crosslinking and immunoprecipitation sequencing; cytokine; gene discovery; genome editing; insight; mast cell; novel; omalizumab; public health relevance; response; screening; small molecule inhibitor; tool; transcription factor; transcriptome

PROJECT SUMMARY Asthma is a highly prevalent chronic inflammatory airway disease with tight genetic and mechanistic links to allergy. Hyperproduction of antibodies of the IgE isotype is a hallmark feature of allergy and the “Th2 high” allergic asthma endotype. IgE is produced when B cells undergo class switch recombination (CSR) and differentiate into antibody secreting plasma cells. Secreted IgE arms mast cells and basophils to release inflammatory mediators in response to allergen exposure, fueling both acute anaphylactic responses and chronic inflammation. A therapeutic antibody targeting IgE reduces asthma symptoms and exacerbations in many patients with moderate to severe persistent disease. Understanding the molecular programming that underlies IgE production may lead to novel and more effective approaches for the prevention and treatment of allergy and asthma. MicroRNAs (miRNAs) are tiny regulators of gene expression that mediate powerful biological effects through their concerted action on networks of target mRNAs. Over 100 distinct miRNAs are expressed in B cells, but critical physiological functions have been assigned to few, so far. Several years ago, we developed a robust screening platform for uncovering miRNA regulators of helper T cell differentiation. We have now fully adapted this system for use in primary B cells and used it to uncover miRNA regulators of CSR and IgE production, including miR-221/222, miR-155 and several other strong novel candidates for further study. The central objective of this proposal is to leverage B cell miRNA:target networks to discover genes and pathways involved in the development and pathogenesis of allergy and asthma. Guided by strong preliminary data, we will rigorously interrogate the function of miR-221/222 and select other miRNA families implicated in IgE production using in vitro culture systems and mouse aeroallergen exposure models of asthma. We will perform comparative Ago2 HITS-CLIP on B cells and combine this biochemical approach with gene expression and computational analyses to generate a map of experimentally defined functional miRNA binding sites throughout the B cell transcriptome. These data will facilitate miRNA-directed discovery of genes and pathways involved in IgE production. Based on preliminary data, we have selected the transcription factor Foxp1 and two other novel miR-221/222 targets to be the first subjects of detailed analysis, taking advantage of existing mutant mice, small molecule inhibitors, and CRISPR/Cas9 genome editing protocols in mouse and human primary B cells. We expect the proposed research to generate novel insights about allergic sensitization and the immunopathogenesis of allergic asthma. In addition, our studies will advance our fundamental understanding of the network properties of miRNA regulation of gene expression and cell behavior.

项目总结 哮喘是一种高度流行的慢性炎症性呼吸道疾病，与 过敏。IgE同型抗体的过度产生是过敏和“Th2兴奋”的一个显著特征 过敏性哮喘内型。当B细胞经历类交换重组(CSR)和 分化为分泌抗体的浆细胞。分泌的IgE臂肥大细胞和嗜碱性粒细胞释放 炎症介质对过敏原暴露的反应，刺激急性过敏反应和 慢性炎症。靶向IgE的治疗性抗体可减少哮喘症状和加重 许多患者患有中到重度的持续性疾病。理解分子编程 免疫球蛋白E的产生可能导致新的和更有效的方法来预防和治疗 过敏和哮喘。 MicroRNAs(MiRNAs)是基因表达的微小调节器，介导强大的生物效应 通过它们对目标mRNAs网络的协调行动。超过100个不同的miRNAs在B中表达 细胞，但到目前为止，关键的生理功能被分配给很少的人。几年前，我们开发了一种 强大的筛选平台，用于揭示辅助T细胞分化的miRNA调节因子。我们现在已经完全 将该系统用于原代B细胞，并用它揭示了CSR和IgE的miRNA调节因子 生产，包括miR-221/222，miR-155和其他几个强有力的候选小说进一步研究。 这项提议的中心目标是利用B细胞miRNA：靶向网络来发现基因和 过敏症和哮喘的发生发展和发病机制。以强劲的前期工作为指导 数据，我们将严格询问miR-221/222的功能，并选择与 使用体外培养系统和哮喘小鼠空气变应原暴露模型生产免疫球蛋白。我们会 在B细胞上进行比较Ago2 HITS-CLIP，并将这种生化方法与基因表达相结合 和计算分析，以生成实验定义的功能性miRNA结合位点图 在整个B细胞转录组中。这些数据将有助于miRNA引导的基因和途径的发现 参与IgE的生产。根据初步数据，我们选择了转录因子Foxp1和两个 其他新型miR-221/222将成为详细分析的第一个主题，利用现有的 突变小鼠、小分子抑制剂和人类和小鼠的CRISPR/Cas9基因组编辑方案 原代B细胞。我们预计拟议的研究将产生关于过敏致敏和 过敏性哮喘的免疫发病机制。此外，我们的学习将促进我们的基础知识 了解miRNA的网络特性对基因表达和细胞行为的调控。