Repression of vascular endothelial growth factor A in glioblastoma cells using engineered zinc finger transcription factors.
Repression of vascular endothelial growth factor A in glioblastoma cells using engineered zinc finger transcription factors.
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发表时间：
2003-12
期刊：
影响因子：
11.2
通讯作者：
A. Snowden;Lei Zhang;F. Urnov;C. Dent;Y. Jouvenot;Xiaohong Zhong;E. Rebar;A. Jamieson;H. S. Zhang;Siyuan Tan;C. Case;C. Pabo;A. Wolffe;P. Gregory
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作者：
A. Snowden;Lei Zhang;F. Urnov;C. Dent;Y. Jouvenot;Xiaohong Zhong;E. Rebar;A. Jamieson;H. S. Zhang;Siyuan Tan;C. Case;C. Pabo;A. Wolffe;P. Gregory
血管生成因子是肿瘤增殖所必需的，因此是有吸引力的治疗靶点。在这项研究中，我们已经使用工程化的锌指蛋白（ZFP）转录因子（TF）抑制血管内皮生长因子（VEGF）-A在人癌细胞系中的表达。我们通过将针对VEGF-A启动子的设计ZFP与甲状腺激素受体α或其病毒相对物vErbA的配体结合结构域融合来创建有效的转录抑制物。此外，这种ZFP-vErbA阻遏物在体内结合其预期的靶位点，并介导靶向启动子处组蛋白H3和H4的特异性脱乙酰化，这一结果模拟了这些结构域的天然阻遏机制。ZFP介导的VEGF-A抑制的潜在治疗相关性使用高度致瘤性胶质母细胞瘤细胞系U87 MG来解决。尽管该细胞系中VEGF-A异常过表达，但工程化ZFP TF能够将VEGF-A的表达抑制>20倍。在ZFP TF介导的抑制后观察到的VEGF-A水平与非血管生成癌细胞系（U251 MG）的VEGF-A水平相当，这表明用ZFP TF获得的抑制程度足以抑制肿瘤血管生成。因此，工程化的ZFP TF显示为在疾病治疗中具有治疗前景的基因表达的有效调节剂。
Angiogenic factors are necessary for tumor proliferation and thus are attractive therapeutic targets. In this study, we have used engineered zinc finger protein (ZFP) transcription factors (TFs) to repress expression of vascular endothelial growth factor (VEGF)-A in human cancer cell lines. We create potent transcriptional repressors by fusing a designed ZFP targeted to the VEGF-A promoter with either the ligand-binding domain of thyroid hormone receptor alpha or its viral relative, vErbA. Moreover, this ZFP-vErbA repressor binds its intended target site in vivo and mediates the specific deacetylation of histones H3 and H4 at the targeted promoter, a result that emulates the natural repression mechanism of these domains. The potential therapeutic relevance of ZFP-mediated VEGF-A repression was addressed using the highly tumorigenic glioblastoma cell line U87MG. Despite the aberrant overexpression of VEGF-A in this cell line, engineered ZFP TFs were able to repress the expression of VEGF-A by >20-fold. The VEGF-A levels observed after ZFP TF-mediated repression were comparable to those of a nonangiogenic cancer line (U251MG), suggesting that the degree of repression obtained with the ZFP TF would be sufficient to suppress tumor angiogenesis. Thus, engineered ZFP TFs are shown to be potent regulators of gene expression with therapeutic promise in the treatment of disease.