Transgenic Murine Model of Brain Vascular Malformation

描述（由申请人提供）： 该提案的长期目标是探索血管生成在脑中的作用，特别是脑动静脉畸形（BAVM）的作用。我们已经深入研究了手术中取出的BAVM组织。我们将继续这项研究，并开发一种体内动物模型的非肿瘤血管生成相关的脑血管疾病，包括BAVM。用腺病毒载体将基因转移到脑中提供了一种有用的工具，在成年啮齿动物脑中诱导局灶性非肿瘤血管生成。PI已经使用这种技术在小鼠大脑中诱导多种基因的稳定过表达。我们假设腺病毒介导的血管内皮生长因子（VEGF）基因转导到小鼠脑中诱导VEGF过表达。VEGF信号传导与VEGF诱导的促血管生成素/Tie-2信号传导途径上调相结合，刺激局灶性血管网络形成。此外，转化生长因子β-1（TGF-β 1）激活TGF-β 1、ALK-5和Smad 2/3信号通路，促进脑组织中局部血管生成的形成。该提议的具体目的是确定：1）在成年小鼠脑中腺病毒-VEGF（AdVEGF）基因转移后VEGF蛋白质产生是否将增加，以及VEGF的过表达是否将诱导微血管形成的局灶簇; 2）在AdVEGF基因转导后TbR-II、ALK 5和Smad 2/3是否将在TGF-β 1转基因小鼠脑中上调。此外，为了量化内皮细胞增殖和支持细胞分化是否会增加，并最终确定这些新生微血管在TGF-b1和VEGF过表达的小鼠脑中的成熟程度。实验部分包括：1）小鼠中高度可重复的腺病毒载体基因转移; 2）特异性VEGF和TGF-β 1通路检测; 3）使用独特的TGF-β转基因小鼠。结合这些遗传技术可能会导致一个新的，可重复的，有用的动物模型的发展。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障碍的模型，相对较快地导致测试脑血管疾病新治疗方法的能力。

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.