Regulation of Gene Expression in the Th2 Cytokine Locus

描述（由申请人提供）：在Th 2分化期间，Th 2细胞因子基因座经历染色质结构的变化，其促进协调的细胞因子基因表达。各种积极和消极的调控因素合作，使这些变化在一个特定的谱系的方式。本研究的主要目的是了解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。第一个目的是产生一个报告系统，以评估在调节IL-4，IL-5和IL-13的协调表达的Th 2细胞因子基因座内的元素的作用。该报告基因将用于同时评估基因的转录，以及评估顺式元件单独和组合对其调控的作用。第二个目的是创建一个系统，通过该系统可以将Aim 1中的报告基因快速导入小鼠ES细胞。将使用酶介导的盒交换（RMCE）来实现这一目标。适当的Th 2分化对于适当的免疫稳态和抗原应答是至关重要的。失调的T细胞极化已经牵涉到许多病理状态，包括过敏性疾病和自身免疫。从这些研究中收集到的信息将有助于我们理解Th 2分化，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：研究Th 2细胞因子基因表达的调控对于理解免疫介导的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。在这项探索性资助中，我们建议开发一种新的系统来研究Th 2细胞因子基因表达，这将首次全面分析Th 2细胞因子基因座中的调控元件功能。该系统的开发将提供对Th 2细胞因子基因表达的控制的更详细的理解，并最终能够开发能够特异性靶向和调节这些基因表达的治疗方式。

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.