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（申请人提供）：由于未知原因，哮喘的患病率和严重程度一直在增加。尽管我们对病理生理学的理解仍然很差，但人们广泛认为哮喘是一种炎症性疾病，而阐述Th2细胞因子的CD4+ T细胞已被确定为其发育中的主要罪魁祸首。这些基因中的许多基因受到转录后的调节，但其调节尚未得到很好的了解。当前的方法强调了使用微阵列来定义“哮喘特征基因”。尽管这些方法是有用的，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水平与蛋白质产生之间的相关性较差。我们已经开发了一种新的范式，即转录后操纵子假说，该假说指出RNA结合蛋白正在协同调节生物学相关分子的表达，例如参与T细胞激活的分子。我们的中心假设是，在过敏原驱动哮喘中，RNA结合蛋白HUR正在协调调节IL-4和IL-13细胞因子基因。 RNA结合蛋白HUR与MRNA的3'未翻译区域（UTR）中的Au富元素（IS）结合，并调节其稳定性和翻译。在8％的人类基因中发现了基序，在转录后基因表达中起关键作用。特别是，据信IL-4和IL-13的两种主要Th2细胞因子在过敏原驱动的哮喘中起着关键作用，在mRNA稳定性和翻译水平下受到HUR的调节。我们开发了新的方法来识别体内hur mRNA靶标。使用这些方法，我们将IL-4和IL-13确定为HUR目标。我们将以以下两个特定目的检验该假设：1：检查CD4+ T细胞对IL-4和IL-13表达的HUR调制的影响。 2：评估HUR在CD4+ T细胞中在过敏原驱动哮喘模型中的作用。我们的方法将对参与过敏原驱动哮喘的促炎细胞因子基因的调节提供更深入的了解。在转录后水平上更好地了解促炎基因调节可能会导致靶向疗法治疗哮喘。公共卫生相关性：世界各地的过敏和哮喘的兴起继续使医生和科学家困扰。这种增加的原因是未知的。对哮喘治疗的反应不同，很可能是由于患者遗传背景的差异。在转录后基因调节水平上更好地了解哮喘病理生理学将极大地帮助我们理解疾病的发病机理和治疗，并直接影响公共卫生。