Regulation of vascular endothelial growth factor (VEGF) splicing from pro-angiogenic to anti-angiogenic isoforms: a novel therapeutic strategy for angiogenesis.
Regulation of vascular endothelial growth factor (VEGF) splicing from pro-angiogenic to anti-angiogenic isoforms: a novel therapeutic strategy for angiogenesis.
复制标题
DOI：
10.1074/jbc.m109.074930
复制
发表时间：
2010-02-19
期刊：
影响因子：
--
通讯作者：
Bates DO
中科院分区：
文献类型：
--
作者：
Nowak DG;Amin EM;Rennel ES;Hoareau-Aveilla C;Gammons M;Damodoran G;Hagiwara M;Harper SJ;Woolard J;Ladomery MR;Bates DO
血管内皮生长因子（VEGF）是一种促血管生成或抗血管生成的蛋白，取决于末端第8外显子的剪接位点选择。外显子8的近端剪接位点选择（PSS）产生促血管生成异构体，如VEGF165，远端剪接位点选择（DSS）产生抗血管生成异构体，如VEGF165b。细胞对剪接位点选择的决定取决于rna结合剪接因子的活性，如ASF/SF2，这些剪接因子先前已被证明可以调节VEGF剪接位点的选择。为了确定促血管生成剪接位点选择的介导机制，我们在上皮细胞和体内血管生成模型中研究了SR蛋白激酶（SRPK1/2）抑制ASF/SF2磷酸化对剪接位点选择的影响。用胰岛素样生长因子-1 （IGF-1）处理的上皮细胞增加PSS，产生更多的VEGF165和更少的VEGF165b。蛋白激酶C抑制和SRPK1/2抑制抑制了DSS的下调和PSS的增加。IGF-1处理导致ASF/SF2的核定位，并被SPRK1/2抑制所阻断。使用VEGF mRNA序列进行拉下试验和RNA免疫沉淀，鉴定出ASF/SF2结合所需的11个核苷酸序列。在小鼠视网膜新生血管模型中，注射SRPK1/2抑制剂可减少血管生成，这表明调节选择性剪接可能是血管生成病变的一种潜在治疗策略。
Vascular endothelial growth factor (VEGF) is produced either as a pro-angiogenic or anti-angiogenic protein depending upon splice site choice in the terminal, eighth exon. Proximal splice site selection (PSS) in exon 8 generates pro-angiogenic isoforms such as VEGF165, and distal splice site selection (DSS) results in anti-angiogenic isoforms such as VEGF165b. Cellular decisions on splice site selection depend upon the activity of RNA-binding splice factors, such as ASF/SF2, which have previously been shown to regulate VEGF splice site choice. To determine the mechanism by which the pro-angiogenic splice site choice is mediated, we investigated the effect of inhibition of ASF/SF2 phosphorylation by SR protein kinases (SRPK1/2) on splice site choice in epithelial cells and in in vivo angiogenesis models. Epithelial cells treated with insulin-like growth factor-1 (IGF-1) increased PSS and produced more VEGF165 and less VEGF165b. This down-regulation of DSS and increased PSS was blocked by protein kinase C inhibition and SRPK1/2 inhibition. IGF-1 treatment resulted in nuclear localization of ASF/SF2, which was blocked by SPRK1/2 inhibition. Pull-down assay and RNA immunoprecipitation using VEGF mRNA sequences identified an 11-nucleotide sequence required for ASF/SF2 binding. Injection of an SRPK1/2 inhibitor reduced angiogenesis in a mouse model of retinal neovascularization, suggesting that regulation of alternative splicing could be a potential therapeutic strategy in angiogenic pathologies.