Targeting Tumor Angiogenesis with G-Quadruplex Binders

血管内皮生长因子（VEGF）和缺氧诱导因子（HIF-1 α）在缺氧缺血性脑损伤中起着关键作用 在肿瘤血管生成中。本提案的总体目标是探索一种新的治疗策略 目的是通过抑制VEGF和HIF-lcc基因的转录来防止肿瘤血管生成 通过靶向在这些基因的启动子区域中形成的G-四链体结构， 分子。本研究的具体目的如下：（1）研究G-四链体是否 在肿瘤细胞中，在VEGF和HIF-1 α基因的启动子区域中体内形成结构， 以小分子为目标。(2)为了确定G-四链体结构在VEGF中的作用， HIF-la启动子区在这些基因的调控中如何与G-四链体化合物相互作用 调节这些基因的调节。(3)为了探索G-四链体的潜在应用， 相互作用剂作为一类新的抗血管生成剂，可以抑制转录活性， VEGF和HIF-1 α基因的表达。(4)发现新的G-四链体化合物或新的铅 基于已知类别的G-四链体相互作用剂的化合物，其特异性抑制 通过VEGF和FHF-la基因的启动子活性介导的转录激活。 在VEGF和HIF-1 α基因的启动子区域中G-四链体结构的形成将被抑制。 通过使用体内化学和酶探测技术进行研究。我们将使用突变分析， 启动子报告基因测定，以确定在启动子区域中形成的G-四链体结构的作用 这些基因。利用多重RT-PCR、微阵列分析、ChIP检测、生物发光成像 技术和血管生成测定，我们将评估体外和体内靶向选择性， G-四链体相互作用剂的抗血管生成活性。最后，无细胞生物化学测定和细胞生物化学测定。 基于高通量筛选系统将用于识别其他先导化合物， 下调细胞培养系统中VEGF或HIF-1 α的表达。 这些研究直接导致了G-100的抗血管生成和抗肿瘤作用的评价。 在人肾癌的临床前模型中的四重相互作用剂， 由于von Hippel Lindau的频繁丢失，HIF-1 α和VEGF的异常过度表达， (VHL)肿瘤抑制基因

The vascular endothelial growth factor (VEGF) and hypoxia induciblefactor (HIF)-la play a pivotal role in tumor angiogenesis. The overall objective of this proposal is to explore a new therapeutic strategy aimed at preventing tumor angiogenesis by repressing the transcription of VEGF and HIF-lcc genes through targeting G-quadruplex structures formed in the promoter region of these genes with small molecules. The specific aims of this proposal are as follows: (1) To investigate whether G-quadruplex structures are formed in vivo in the promoter regions of VEGF and HIF-la genes in tumor cells and can be targeted with small molecules. (2) To determine the role of G-quadruplex structures in VEGF and HIF-la promoter regions in the regulation of these genes and how G-quadruplex interactive compounds modulate the regulation of these genes. (3) To explore the potential application of G-quadruplex- interactive agents as a new class of anti-angiogenic agents, which can repress the transcriptional activity of the VEGF and HIF-la genes. (4) To discover new classes of G-quadruplex compounds or new lead compounds based on known classes of G-quadruplex-interactive agents that specifically inhibit the transcriptional activation mediated by the promoter activity of VEGF and FHF-la genes. The formation of G-quadruplex structures in the promoter region of the VEGF and HIF-la genes will be studied by using in vivo chemical and enzymatic probing techniques. We will use mutation analysis and promoter reporter assays to determine the role of G-quadruplex structures formed in the promoter region of these genes. Utilizing multiple RT-PCR, microarray analysis, ChIP assay, bioluminescent imaging technique, and angiogenesis assays, we will evaluate in vitro and in vivo target selectivity and antiangiogenic activity of G-quadruplex-interactive agents. Lastly, cell free biochemical assays and cell based high-throughput screening system will be utilized to identify other lead compounds that selectively downregulate either VEGF or HIF-la expression in a cell culture system. These studies lead directly to the evaluation of the anti-angiogenic and antitumor effects of G- quadruplex-interactive agents in a preclinical model of human kidney cancer, a tumor type characterized by the abnormal overexpression of HIF-la and VEGF due to the frequent loss of the von Hippel Lindau (VHL) tumor suppressor gene.