Posttranscriptional Gene Regulation in Asthma

哮喘的转录后基因调控

基本信息

批准号：
7847589
负责人：
ULUS ATASOY
金额：
$ 17.87万
依托单位：
UNIVERSITY OF MISSOURI-COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-22 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7847589
关键词：
3&apos Untranslated Regions Affect Allergens Allergic inflammation Animal Model Animals Area Asthma Binding Biological Biological Assay Boxing CD4 Positive T Lymphocytes Cell Nucleus Cloning Complex Cytokine Gene Cytoplasm Data Development Disabled Persons Disease ERG gene Elements Employee Strikes Extrinsic asthma Functional disorder Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Goals Human Hypersensitivity IL13 gene Immunoprecipitation Indium Inflammation Inflammatory Inflammatory Response Interferons Interleukin-13 Interleukin-4 Knock-out Lead Lentivirus Vector Messenger RNA Methods Modality Modeling Mus Operon Outcome Ovalbumin Ovum Pathogenesis Patients Phenotype Physicians Play Prevalence Production Proteins Public Health Publishing RNA-Binding Proteins Regulation Reporting Research Research Personnel Role Scientist Severities T-Cell Activation Testing Therapeutic Time Tissues Translations Untranslated Regions Up-Regulation Work allergic airway inflammation base cDNA Arrays chemokine cytokine design experience handicapping condition in vivo innovation loss of function mRNA Stability novel novel strategies public health relevance response

项目摘要

DESCRIPTION (provided by applicant): Asthma has been increasing in prevalence and severity for unknown reasons. Though our understanding of the pathophysiology remains poor, it is widely accepted that asthma is an inflammatory disease and CD4+ T cells elaborating Th2 cytokines have been identified as major culprits in its development. Many of these genes are posttranscriptionally regulated but their regulation is not well understood. Current approaches have emphasized the use of microarrays to define "asthma signature genes". Though these approaches are helpful, they are also incomplete and may miss important target genes since there is a poor correlation between steady-state mRNA levels and protein production. We have developed a new paradigm, the posttranscriptional operon hypothesis, which states that RNA binding proteins are coordinately regulating the expression of biologically related molecules, such as those involved in T cell activation. Our central hypothesis is that the RNA binding protein, HuR, is coordinately regulating IL-4 and IL-13 cytokine genes during allergen driven asthma. The RNA binding protein, HuR, binds to the AU-rich elements (ARE) in the 3' untranslated regions (UTR) of mRNAs and modulates their stability and translation. The ARE motif is found in 8% of human genes and plays a critical role in posttranscriptional gene expression. In particular, two major Th2 cytokines, IL- 4 and IL-13, believed to play critical roles in allergen driven asthma, are regulated by HuR at the level of mRNA stability and translation. We have developed novel methods to identify en masse cellular in vivo HuR mRNA targets. Using these approaches, we have identified IL-4 and IL-13 as HuR targets. We will test this hypothesis with the following two specific aims: 1: Examine the effects of HuR modulation in CD4+ T cells upon IL-4 and IL-13 expression. 2: Assess the role of HuR in CD4+ T cells in allergen driven models of asthma in mice. Our approach will provide a fuller understanding of the regulation of proinflammatory cytokine genes involved in allergen driven asthma. Better understanding of proinflammatory gene regulation at posttranscriptional level may potentially lead to targeted therapies to treat asthma. Public Health Relevance: The rise of allergies and asthma around the world continues to perplex physicians and scientists. The reasons for this increase are unknown. Different responses to therapies for asthma are most likely due to differences in genetic backgrounds of patients. A better understanding of asthma pathophysiology at the posttranscriptional gene regulation level would greatly aid in our understanding of disease pathogenesis and treatment and have a direct impact on public health.

描述（由申请人提供）：由于未知的原因，哮喘的患病率和严重程度不断增加。尽管我们对病理生理学的了解仍然很少，但人们普遍认为哮喘是一种炎症性疾病，而产生 Th2 细胞因子的 CD4+ T 细胞已被确定为其发展的主要罪魁祸首。许多这些基因受到转录后调节，但它们的调节尚不清楚。目前的方法强调使用微阵列来定义“哮喘特征基因”。尽管这些方法很有帮助，但它们也不完整，并且可能会错过重要的靶基因，因为稳态 mRNA 水平和蛋白质产生之间的相关性很差。我们开发了一种新的范式，即转录后操纵子假说，该假说指出 RNA 结合蛋白协调调节生物相关分子的表达，例如那些参与 T 细胞激活的分子。我们的中心假设是，RNA 结合蛋白 HuR 在过敏原驱动的哮喘过程中协调调节 IL-4 和 IL-13 细胞因子基因。 RNA 结合蛋白 HuR 与 mRNA 3' 非翻译区 (UTR) 中富含 AU 的元件 (ARE) 结合，并调节其稳定性和翻译。 ARE 基序存在于 8% 的人类基因中，在转录后基因表达中发挥着关键作用。特别是，两种主要的 Th2 细胞因子 IL-4 和 IL-13 被认为在过敏原驱动的哮喘中发挥关键作用，它们在 mRNA 稳定性和翻译水平上受到 HuR 的调节。我们开发了新的方法来识别体内大量细胞 HuR mRNA 靶标。使用这些方法，我们已确定 IL-4 和 IL-13 作为 HuR 靶标。我们将通过以下两个具体目标来检验这一假设： 1：检查 CD4+ T 细胞中 HuR 调节对 IL-4 和 IL-13 表达的影响。图 2：评估 HuR 在过敏原驱动的小鼠哮喘模型中的 CD4+ T 细胞中的作用。我们的方法将提供对参与过敏原驱动的哮喘的促炎细胞因子基因的调节的更全面的了解。更好地了解转录后水平的促炎基因调控可能会导致治疗哮喘的靶向疗法。公共卫生相关性：世界各地过敏和哮喘发病率的上升继续困扰着医生和科学家。这种增加的原因尚不清楚。对哮喘治疗的不同反应很可能是由于患者遗传背景的差异造成的。在转录后基因调控水平上更好地了解哮喘病理生理学将极大地有助于我们了解疾病发病机制和治疗，并对公众健康产生直接影响。