Targeting Tumor Angiogenesis with G-Quadruplex Binders

使用 G-四联体结合剂靶向肿瘤血管生成

• 批准号: 7772384
• 负责人: DAEKYU SUN
• 金额: $ 26.03万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2012-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7772384
• 关键词: Angiogenesis Inhibitors; Binding; Biochemical; Biochemistry; Biological Assay; Blood capillaries; Cell Culture System; Cells; Cellular biology; Chemicals; Clinical Research; DNA Structure; Drug Delivery Systems; Elements; Endothelial Cells; Evaluation; G-Quartets; Gene Expression Regulation; Gene Structure; Genes; Genetic Transcription; Genitourinary system; Growth Factor; Human; Hypoxia; Imaging Techniques; In Vitro; Lead; Malignant Neoplasms; Mediating; Microarray Analysis; Migration Assay; Mutation Analysis; Nude Mice; Oncologist; Permeability; Play; Pre-Clinical Model; Principal Investigator; Process; Promoter Regions; Regulation; Renal carcinoma; Reporter; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Role; Structure; System; Techniques; Testing; The Sun; Therapeutic; Transcriptional Activation; Tube; Tumor Angiogenesis; Tumor Suppressor Genes; Vascular Endothelial Growth Factors; angiogenesis; base; capillary; cytokine; drug development; high throughput screening; human VEGF protein; hypoxia inducible factor 1; in vivo; interest; migration; neoplastic cell; novel therapeutics; overexpression; pre-clinical; prevent; programs; promoter; protein function; response; small molecule; transcription factor; tumor; tumor xenograft

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.

血管内皮生长因子(VEGF)和缺氧诱导因子-1α(HIF-1α)在其中起着关键作用 在肿瘤血管生成中的作用。这项提议的总体目标是探索一种新的治疗策略 通过抑制血管内皮生长因子和缺氧诱导因子-LCC基因的转录来预防肿瘤血管生成 通过靶向这些基因启动子区域形成的G-四链结构 分子。这项建议的具体目的如下：(1)调查G-四联体 肿瘤细胞中VEGF和HIF-1a基因的启动子区域在体内形成结构，并可以 以小分子为靶标。(2)确定G-四链结构在血管内皮生长因子和血管内皮生长因子中的作用。 HIF-1a启动子区域对这些基因的调控以及G-四链化合物是如何相互作用的 调节这些基因的调节。(3)探索G-四链的潜在应用。 相互作用剂作为一类新型的抗血管生成药物，可以抑制转录活性 血管内皮生长因子和缺氧诱导因子-1a基因。(4)发现新的G-四链体化合物或新的先导化合物 基于已知类别的G-四链相互作用剂的化合物，这些化合物特别抑制 血管内皮生长因子和FHF-1a基因启动子活性介导的转录激活。 在VEGF和HIF-1a基因启动子区域形成G-四链结构将是 通过体内化学和酶探测技术进行研究。我们将使用突变分析和 启动子报告分析确定启动子区域形成的G-四链结构的作用 这些基因。利用多重RT-PCR、微阵列分析、芯片分析、生物发光成像 技术和血管生成试验，我们将评估体外和体内靶向选择性和 G-四链相互作用剂的抗血管生成活性。最后，无细胞生化分析和细胞 基于高通量的筛选系统将被用来鉴定其他选择性地 下调细胞培养系统中血管内皮生长因子或缺氧诱导因子-1α的表达。 这些研究直接导致对G-半乳糖抗血管生成和抗肿瘤作用的评价。 四链相互作用药物在人类肾癌临床前模型中的作用 由于von Hippel Lindau的频繁丢失导致HIF-1α和VEGF的异常过度表达 (VHL)肿瘤抑制基因。