Transgenic Murine Model of Brain Vascular Malformation

描述(由申请人提供)： 这项建议的长期目标是探索血管生成在大脑中的作用，特别是脑动静脉畸形(BAVM)的作用。我们对手术中切除的脑动静脉畸形组织进行了深入的研究。我们将继续这项研究，并开发一种与脑血管疾病相关的非肿瘤血管生成的体内动物模型，包括脑动静脉畸形。用腺病毒载体将基因转移到脑内提供了一种在成年啮齿动物脑内诱导局部、非肿瘤血管生成的有用工具。PI已经使用这项技术在小鼠大脑中诱导了各种基因的稳定过度表达。我们假设，腺病毒介导的血管内皮生长因子(VEGF)基因导入小鼠脑内可诱导VEGF过表达。血管内皮生长因子信号与血管生成素/Tie-2信号通路的上调结合，刺激局部血管网络的形成。此外，转化生长因子β-1(转化生长因子-β1)激活转化生长因子-β1、ALK-5和Smad2/3信号通路，促进脑组织局部血管生成。这一建议的具体目的是确定：1)腺病毒-血管内皮生长因子(AdVEGF)基因转移是否会增加成年小鼠脑内VEGF蛋白的产生，以及VEGF的过表达是否会导致局部微血管形成；2)AdVEGF基因转导后，Tbr-11、Alk5和Smad2/3是否会在转化生长因子-β1转基因小鼠脑中上调。此外，为了量化是否会增加内皮细胞的增殖和支持细胞分化，最后，确定这些新的微血管在转化生长因子-β1和血管内皮生长因子过表达的小鼠脑中的成熟程度。实验部分包括：1)高重复性的腺病毒载体在小鼠体内的基因转移；2)特异性的血管内皮生长因子和转化生长因子-β1途径的检测；以及3)使用独特的转化生长因子-b转基因小鼠。将这些基因技术结合起来，可能会开发出一种新颖的、可重复的、有用的动物模型。这种非肿瘤血管生成啮齿动物制剂将作为人类脑血管生成紊乱的模型，从而相对快速地测试脑血管疾病的新治疗方法。

DESCRIPTION (provided by applicant): The long-term objectives of this proposal are to explore the roles of angiogenesis in the brain, especially the role of brain arteriovenous malformations (BAVMs). We have intensively studied BAVM tissue removed during surgery. We will continue this study and develop an in vivo animal model of non-tumor angiogenesis relevant to cerebrovascular diseases including BAVMs. In vivo gene transfer into the brain with adenoviral vectors provides a useful tool inducing focal, non-tumor angiogenesis in the adult rodent brain. The PI has used this technique to induce stable overexpression of a variety of genes in the mouse brain. We hypothesize that adenoviral-mediated vascular endothelial growth factor (VEGF) gene transduction into the mouse brain induces VEGF overexpression. VEGF signaling, combined with VEGF-induced upregulation of the angiopoietin/Tie-2 signaling pathway, stimulates focal vascular network formation. Furthermore, transforming growth factor beta-1 (TGF-b1) activates TGF-b1, ALK-5, and Smad 2/3 signal pathways, promoting regional angiogenesis formation in the brain tissue. The specific aims of this proposal are to determine: 1) whether VEGF protein production will be increased following adenoviral-VEGF (AdVEGF) gene transfer in the adult mouse brain, and whether overexpression of VEGF will induce focal clusters of microvasculature formation; 2) whether TbR-ll, ALK5, and Smad 2/3 will be upregulated in the TGF-bl transgenic mouse brain following AdVEGF gene transduction. Furthermore, to quantify whether endothelial cell proliferation and support cell differentiation will be increased, and finally, to identify how mature these neo-microvasculature are in the mouse brain with both TGF-b1 and VEGF overexpression. The experimental components include: 1) highly reproducible adenoviral vector gene transfer in mice; 2) specific VEGF and TGF-b1 pathway detection; and 3) using unique TGF-b transgenic mice. Combining these genetic techniques may lead to the development of a novel, reproducible, and useful animal model. This non-tumor angiogenesis rodent preparation will serve as a model of disordered human cerebral angiogenesis, leading relatively quickly to the ability to test new therapeutic approaches for cerebrovascular disorders.