MicroRNA directed pathway discovery in allergy and asthma

MicroRNA 指导过敏和哮喘通路的发现

基本信息

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
项目类别：
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

项目摘要

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.

项目概要哮喘是一种高度流行的慢性炎症性气道疾病，与遗传和机制密切相关。 IgE 同种型抗体的过度产生是过敏和“Th2 高”的标志特征。当 B 细胞进行类别转换重组 (CSR) 时，会产生过敏性哮喘内型 IgE。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作出反应，加剧急性过敏反应和一种针对 IgE 的治疗性抗体可减轻哮喘症状和恶化。许多患有中度至重度持续性疾病的患者了解分子编程。 IgE 产生的基础可能会带来新的、更有效的方法来预防和治疗过敏和哮喘。 MicroRNA (miRNA) 是基因表达的微小调节因子，可介导强大的生物效应通过它们对目标 mRNA 网络的协同作用，B 中表达了 100 多种不同的 miRNA。细胞，但关键的生理功能已分配给少数细胞，几年前，我们开发了一种。我们现在已经完全拥有用于发现辅助 T 细胞分化的 miRNA 调节因子的强大筛选平台。将该系统改造用于原代 B 细胞，并用它来发现 CSR 和 IgE 的 miRNA 调节因子生产，包括 miR-221/222、miR-155 和其他几个有待进一步研究的强大新候选者。该提案的核心目标是利用 B 细胞 miRNA：目标网络来发现基因和涉及过敏和哮喘的发展和发病机制的途径受到强有力的初步指导。数据，我们将严格询问 miR-221/222 的功能并选择其他涉及的 miRNA 家族我们将使用体外培养系统和小鼠气源性过敏原暴露模型来生产 IgE。对 B 细胞进行比较 Ago2 HITS-CLIP，并将这种生化方法与基因表达结合起来和计算分析生成实验定义的功能性 miRNA 结合位点图谱这些数据将有助于 miRNA 指导的基因和通路的发现。根据初步数据，我们选择了Foxp1和两个转录因子。其他新的 miR-221/222 靶点将成为详细分析的首要对象，利用现有的突变小鼠、小分子抑制剂以及小鼠和人类的 CRISPR/Cas9 基因组编辑方案我们期望拟议的研究能够产生关于过敏致敏和此外，我们的研究将推进我们的基础研究。了解 miRNA 调节基因表达和细胞行为的网络特性。