Forkhead signaling in the endothelium

描述（由申请方提供）：内皮细胞（EC）表型在健康和疾病中显示出显著的异质性。血管生物学的一个重要目标是了解EC表型的时空调控的分子机制。这种竞争性的更新申请是我们以前工作的延伸，旨在阐明叉头（FOXO）转录因子在EC生物学中的作用。在过去的3年里，FOXO生物学出现了三个新颖而令人兴奋的方面，这些方面与我们自己发表的数据和初步结果一起为当前的提议提供了坚实的基础：1）尽管FOXO蛋白普遍表达，但FOXO缺陷型小鼠显示血管表型，无论该基因在胚胎发生期间或在成人中缺失，2）血管表型是器官特异性的，并且来自不同器官的FOXO缺陷EC表现出不同的基因表达模式和功能变化，以及3）FOXO蛋白（至少在其他细胞类型中）已显示与广谱转录因子结合和/或相互作用。基于这些考虑，我们假设FOXO蛋白（特别是FOXO 1）与其他转录因子合作调节EC表型。总体目标是确定FOXO 1相关的转录网络在内皮细胞，了解这些如何介导血管床特异性基因表达，并确定FOXO 1在完整的内皮细胞中的体内作用。目的1是研究FOXO 1与内皮细胞中其他转录因子的相互作用。目的二是研究FOXO 1和NF-κ B在内皮细胞中的组合基因调控。目的3分析FOXO 1条件性过表达和敲除对正常和病理状态下EC功能和功能障碍的影响。

DESCRIPTION (provided by applicant): Endothelial cell (EC) phenotypes display remarkable heterogeneity in health and disease. An important goal in vascular biology is to understand the molecular mechanisms underlying the spatial and temporal modulation of EC phenotypes. This competing renewal application is an extension of our previous work aimed towards elucidating the role of forkhead (FOXO) transcription factors in EC biology. Three novel and exciting aspects of FOXO biology have emerged in the past 3 years, which together with our own published data and preliminary results provide a strong foundation for the current proposal: 1) despite the fact that FOXO proteins are ubiquitously expressed, FOXO- deficient mice display a vascular phenotype, whether the gene is deleted during embryogenesis or in adults, 2) the vascular phenotype is organ-specific and FOXO-deficient ECs from different organs demonstrate different gene expression patterns and functional changes, and 3) FOXO proteins (at least in other cell types) have been shown to bind to and/or interact with a broad spectrum of transcription factors. Based on these considerations, we hypothesize that FOXO proteins (in particular FOXO1) cooperate with other transcription factors to regulate EC phenotypes. The overall goal is to identify FOXO1-associated transcriptional networks in ECs, to understand how these mediate vascular bed-specific gene expression, and to define the in vivo role of FOXO1 in the intact endothelium. Aim 1 is to characterize functional interactions between FOXO1 and other transcription factors in ECs. Aim 2 is to characterize combinatorial gene regulation by FOXO1 and NF-kB in ECs. Aim 3 is to analyze the effect of conditional FOXO1 over-expression and knockout on EC function and dysfunction in normal and pathological states.