SWI/SNF-REGULATED POSTNATAL ANGIOGENESIS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 依赖于ATP的染色质重塑复合体，如哺乳动物的SWI/SNF复合体，促进了基因表达的适当的时空模式，因此在许多发育过程中发挥着重要作用。我们以前已经证明了SWI/SNF在胚胎血管发育中的作用，现在建议将我们的研究扩展到成人。我们假设，当使用可诱导的内皮特异性VE-钙粘连蛋白-Creer转基因小鼠从出生后的内皮细胞中删除时，漂浮的SWI/SNF催化亚基-BRG1-将产生重要的表型。我们建议研究出生后视网膜的生理性血管发育和肿瘤模型中的病理血管发育，重点是与内皮BRG1缺失的胚胎在表型上的相似或不同。基于我们的初步数据，我们将使用体内和体外分析来解决特定的假设，即在缺乏血管BRG1的情况下，Wnt信号是错误调节的。最后，我们建议利用SWI/SNF染色质重塑复合体通过与靶基因的调控元件物理相互作用来介导其表型效应这一事实来确定基因组靶标。我们将筛选对照细胞和突变细胞，寻找血管生成和Wnt信号阵列以及无偏见全基因组阵列上所代表的基因的表达变化。总之，我们提出的实验，结合我们对SWI/SNF在胚胎血管发育中的作用的初步了解，将有助于阐明血管生成的表观遗传调控，并揭示这一重要发育过程的新介质。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. ATP-dependent chromatin-remodeling complexes, such as the mammalian SWI/SNF complexes, facilitate proper temporal and spatial patterns of gene expression and therefore play important roles in a number of developmental processes. We have previously demonstrated a role for SWI/SNF in embryonic vascular development and are now proposing to extend our studies into the adult. We hypothesize that the floxed SWI/SNF catalytic subunit--Brg1--will yield important phenotypes when deleted from postnatal endothelium using inducible, endothelial-specific VE-Cadherin-CreER transgenic mice. We propose to investigate both physiological vascular development in the postnatal retina and pathological vascular development in a tumor model, focusing on phenotypic similarities or differences with embryos depleted of endothelial BRG1. Based on our preliminary data, we will address the specific hypothesis that Wnt signaling is misregulated in the absence of vascular BRG1 using both in vivo and in vitro assays. Finally, we propose to identify genomic targets of SWI/SNF chromatin-remodeling complexes by taking advantage of the fact that the complexes mediate their phenotypic effects by physically interacting with regulatory elements of target genes. We will screen control versus mutant cells for expression changes in genes represented on angiogenesis and Wnt signaling arrays as well as unbiased whole-genome arrays. Altogether, our proposed experiments, in combination with our preliminary understanding of the role of SWI/SNF in embryonic vascular development, stand to elucidate the epigenetic regulation of angiogenesis and to reveal novel mediators of this important developmental process.