Systematic Responses of GILZ Regulatory Network to Alcohol

描述（由申请人提供）：长期以来，酒精与抗炎症和免疫抑制有关。这些效应背后的分子机制尚未完全确定。在初步研究中，我们对暴露于不同剂量酒精的人气道上皮进行了全基因组高密度微阵列分析。鉴定出一组酒精反应基因。其中，糖皮质激素诱导的亮氨酸拉链基因（GILZ）是一个重要的糖皮质激素（GC）靶基因，其对酒精的反应呈剂量依赖性。GILZ是细胞中转导糖类固醇作用的主要介质。这一新发现揭示了一个潜在的重要信号通路，可能有助于酒精相关的抗炎症和免疫抑制。本应用程序的最终目标是在系统级别定义酒精特异性GILZ调节网络。有三个具体目的：1)具体目的1：确定与GILZ基因启动子结合并负责酒精介导的GILZ激活的反式作用因子；特异性目的2：通过染色质免疫沉淀测序（ChIP-seq）检测酒精诱导的GILZ染色体占用，以分析整个基因组中的GILZ靶向序列，并描绘酒精特异性GILZ分子网络；3)特异性目的3：明确酒精暴露下多类型细胞对LPS的炎症细胞因子反应中GILZ的作用。我们提出的研究将允许我们验证酒精诱导的GILZ上调通过一个特定的调节网络有助于酒精抗炎症和免疫抑制的中心假设。通过完成这一提议，我们将获得突破性的知识，以帮助理解酒精如何像类固醇一样在分子上调节细胞功能。本研究将对酒精诱导的抗炎和免疫抑制机制提供新的见解，这可能有助于合理设计治疗酒精相关疾病的治疗方法。

DESCRIPTION (provided by applicant): Alcohol has long been associated with anti-inflammation and immunosuppression. The molecular mechanisms underlying these effects are not fully defined. In the preliminary studies, we performed whole-genome high-density microarray analyses on human airway epithelia exposed to varied doses of alcohol. A cluster of alcohol-responsive genes was identified. Among them, glucocorticoid-induced leucine zipper (GILZ), a prominent glucocorticoid (GC) target gene, responded to alcohol in a dose-dependent manner. GILZ is a major mediator to transduce glucosteroid actions in cells. This novel finding reveals a potentially important signaling pathway which may contribute to the alcohol-associated anti-inflammation and immunosuppression. The ultimate goal of this application is to define alcohol-specific GILZ regulatory network at the system level. There are three specific aims: 1) Specific Aim 1: To identify the trans-acting factors which bind to the GILZ gene promoter and are responsible for alcohol-mediated activation of GILZ; Specific Aim 2: To examine GILZ chromosomal occupancy induced by alcohol by chromatin immunoprecipitation sequencing (ChIP-seq) to profile the GILZ-targeting sequences across the genome and to delineate the alcohol-specific GILZ molecular network; 3) Specific Aim 3: To define the GILZ role in inflammatory cytokine response to LPS in multiple types of cells under alcohol exposure. The proposed research will allow us to test the central hypothesis that the alcohol-induced GILZ upregulation contributes to alcohol anti- inflammation and immunosuppression through a specific regulatory network. By completion of this proposal, we will gain the groundbreaking knowledge to help understand how alcohol acts like steroids to modulate cellular functions molecularly. This research will provide new insights into the mechanism of alcohol-induced anti-inflammation and immunosuppression, which may lead to rational design of therapeutics for the treatment of alcohol-associated diseases.