Regulation of Gene Expression in the Th2 Cytokine Locus

描述(申请人提供)：在Th2分化过程中，Th2细胞因子基因座经历染色质结构的变化，从而促进细胞因子基因的协调表达。各种积极和消极的调节因素协同工作，以特定血统的方式带来这些变化。这项研究的主要目的是了解在该基因座内协调基因表达的顺式元件之间的功能相互作用。我们还想了解转录因子启动和维持细胞因子基因转录活性的机制。第一个目标是建立一个报告系统，以评估Th2细胞因子基因座内的元件在调节IL-4、IL-5和IL-13协调表达方面的作用。这位记者将被用来同时评估基因的转录，以及评估顺式元件单独和组合在其调控中的作用。第二个目标是创建一种系统，通过该系统可以将目标1中的报告基因快速导入小鼠ES细胞。重组酶介导的盒式磁带交换(RMCE)将被用于实现这一目标。适当的Th2分化对于适当的免疫动态平衡和抗原反应性至关重要。调节失调的T细胞极化与许多病理状态有关，包括变态反应性疾病和自身免疫。从这些研究中收集的信息将有助于我们对Th2分化的理解，并有望实现最终目标，干预这一过程，避免或纠正这些病理表现。公共卫生相关性：对Th2细胞因子基因表达调控的研究对于了解免疫介导性功能障碍(如哮喘或自身免疫性疾病)的发展和治疗至关重要。在这项探索性拨款中，我们建议开发一种新的系统来研究Th2细胞因子基因的表达，这将首次使全面分析Th2细胞因子基因座中的调节元件功能成为可能。这一系统的开发将提供对Th2细胞因子基因表达控制的更详细的了解，并最终将使能够特异性靶向和调节这些基因表达的治疗方式的开发成为可能。

DESCRIPTION (provided by applicant): During Th2 differentiation, the Th2 cytokine locus undergoes changes in chromatin structure that facilitate coordinated cytokine gene expression. A variety of positive and negative regulatory elements work cooperatively to bring about these changes in a lineage-specific fashion. The principal goal of this study is to understand the functional interplay among cis-elements that coordinates gene expression within this locus. We also want to understand the mechanism by which transcription factors initiate and maintain transcriptional activity of the cytokine genes. The first aim is to generate a reporter system to evaluate the role of elements within the Th2 cytokine locus in regulating coordinated expression of IL-4, IL-5, and IL-13. This reporter will be utilized to assess transcription of the genes simultaneously as well as assess the role of cis-elements, individually and in combination, on their regulation. The second aim is to create a system by which the reporter in Aim 1 can be rapidly introduced into mouse ES cells. Recombinase-mediated cassette exchange (RMCE) will be used to fulfill the goals of this aim. Appropriate Th2 differentiation is critical for proper immune homeostasis and antigen responsiveness. Dysregulated T cell polarization has been implicated in a number of pathological states, including allergic diseases and autoimmunity. Information gleaned from these studies will contribute to our understanding of Th2 differentiation and hopefully enable the ultimate goal, to intervene in this process and to avoid or correct these pathological manifestations. PUBLIC HEALTH RELEVANCE: The study of the regulation of Th2 cytokine gene expression is crucial in order to understand the development and treatment of immune-mediated dysfunction such as asthma or autoimmune disease. In this exploratory grant, we propose to develop a novel system to study Th2 cytokine gene expression that will enable for the first time a comprehensive analysis of regulatory element function in the Th2 cytokine locus. The development of this system will provide a much more detailed understanding of the control of Th2 cytokine gene expression and will eventually enable the development of therapeutic modalities that can specifically target and modulate the expression of these genes.