G-quadruplexes formed in human oncogene promoters and their drug complexes

人类癌基因启动子及其药物复合物中形成的G-四链体

• 批准号: 7836507
• 负责人: DANZHOU YANG
• 金额: $ 2.1万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-03 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7836507
• 关键词: Address; Affinity Chromatography; Antineoplastic Agents; Binding; Binding Proteins; Biological; Biological Assay; Biological Factors; Biological Process; Cell Line; Complex; Crystallization; DNA; DNA Binding; DNA Structure; Data; Development; Drug Controls; Drug Delivery Systems; Drug Design; Drug Interactions; Elements; Equilibrium; Evaluation; Fluorescence Resonance Energy Transfer; Funding; Future; G-Quartets; G-substrate; Gene Expression; Genes; Genetic Transcription; Goals; Grant; Growth; Human; Hypersensitivity; In Vitro; Isomerism; Laboratories; Malignant Epithelial Cell; Malignant Neoplasms; Methods; Molecular; Molecular Conformation; Molecular Models; Molecular Structure; Molecular Target; Mutate; NMR Spectroscopy; Nuclear; Oncogenes; Pathway interactions; Pharmaceutical Preparations; Phase II Clinical Trials; Physiological; Plasmids; Play; Primary carcinoma of the liver cells; Promoter Regions; Protein Binding; Proteins; Proteomics; Recovery; Reporting; Research; Role; Screening procedure; Single-Stranded DNA; Solutions; Specificity; Structure; System; Therapeutic Intervention; Thermodynamics; Time Study; Transcriptional Activation; Transcriptional Regulation; Transcriptional Silencer Elements; United States National Institutes of Health; Vascular Endothelial Growth Factors; Work; abstracting; base; c-myc Genes; cancer therapy; design; drug development; in vivo; insight; microcalorimetry; molecular modeling; mutant; novel; outcome forecast; overexpression; parent grant; promoter; response; small molecule; therapeutic target; tool; tumor

DESCRIPTION: This application is in response to the Notice Number (NOT-OD-09-058) and Notice Title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications. G-quadruplex DNA secondary structures have recently emerged as a new class of cancer-specific molecular targets for anticancer drugs. There is compelling in vitro and in vivo evidence of G-quadruplex structures formed in the promoter regions of genes involved in growth and proliferation as transcriptional regulators. Our parent grant proposed to define the molecular structures of the G-quadruplexes in the promoter regions of the four oncogenes, c-Myc, bcl-2, VEGF, and HIF-1a, and their drug-complex(es). While the DNA G-quadruplexes are very promising new drug targets, the evaluation of their potential as cancer therapeutic targets depends on the understanding of biologically relevant G-quadruplex structures. To that end, it appears that a number of additional important questions need to be addressed in order to fully understand the biological roles of the promoter Gquadruplexes and to evaluate their potential as cancer therapeutic targets, in particular, the specific protein interactions of the G-quadruplex structures and the consideration of the intact molecular system of the DNA secondary structures formed in gene promoters. Thus we propose in this Revision Application to study these two important expanded aspects. The proposed research in the Revision Application will focus on the c-Myc promoter as it is the most extensively studied system. In particular, the c-Myc promoter contains a highly conserved polyG/polyC NHE III1, which controls 80-90% of the c-Myc transcription. The NHE III1 has been shown to be a silencer element and the formation of a DNA G-quadruplex structure is critical for c-Myc transcriptional silencing. Furthermore, compounds that stabilize this G-quadruplex conformation have been shown to reduce c-Myc expression and are antitumorigenic. Specifically, we propose: 1) to identify the protein(s) that bind the DNA G-quadruplex formed in the c-Myc promoter; and 2) to define the structure of the Imotif as a second target in the silencer element of the c-Myc promoter. The two specific aims are important expansions of the parent grant. Protein interactions and functions will be essential to fully understand the biological function of the DNA secondary structures in gene promoters, and the identification of the G4 interactive protein(s) is the first step towards that goal. By including the I-motif formed in the C-rich strand, the revision will expand the scope of the parent grant to the complete molecular system of the silencer element in the c-Myc promoter for drug targeting. Our long-term objective is to use a structure-based approach to rationally design small molecule compounds that specifically bind DNA secondary structures and modulate gene transcription. The Revision Application will clearly accelerate the research on targeting DNA secondary structures formed in the gene promoter regions for anticancer drug development. The two specific aims are not highly inter-dependent and both have a high likelihood of being completed in the two-year period. PUBLIC HEALTH RELEVANCE: c-Myc is an important oncogene and has been validated as potential targets for cancer therapeutic intervention. It is overexpressed in a variety of tumor types and its overexpression has been associated with poor prognosis for survival. Effective modulation of c-myc expression offers promise for the cancer treatment. The proposed research represents a novel new strategy for modulating c-myc gene expression by small molecule drugs.

描述：本申请是对通知号（NOT-OD-09-058）和通知标题的回应：NIH宣布为竞争性修订申请提供恢复法案资金。近年来，g -四重体DNA二级结构已成为一类新的抗癌药物的肿瘤特异性分子靶点。有令人信服的体外和体内证据表明，在参与生长和增殖的基因启动子区域形成的g -四重结构作为转录调节因子。本研究提出确定四种致癌基因c-Myc、bcl-2、VEGF和HIF-1a及其药物复合物启动子区域g-四联体的分子结构。虽然DNA g -四重体是非常有前途的新药物靶点，但评估其作为癌症治疗靶点的潜力取决于对生物学相关g -四重体结构的理解。为此，为了充分了解启动子四重体的生物学作用并评估其作为癌症治疗靶点的潜力，似乎需要解决一些其他重要问题，特别是g -四重体结构的特定蛋白质相互作用以及考虑基因启动子中形成的DNA二级结构的完整分子系统。因此，我们建议在本修订申请中对这两个重要的扩展方面进行研究。由于c-Myc启动子是目前研究最广泛的系统，因此本文拟在Revision Application中重点研究c-Myc启动子。特别地，c-Myc启动子包含一个高度保守的polyG/polyC NHE III1，它控制着80-90%的c-Myc转录。NHE III1已被证明是一个沉默元件，DNA g -四重结构的形成对c-Myc转录沉默至关重要。此外，稳定g -四重构象的化合物已被证明可以减少c-Myc的表达并具有抗肿瘤性。具体来说，我们建议：1)鉴定结合在c-Myc启动子中形成的DNA g -四重体的蛋白质；以及2)将immotif的结构定义为c-Myc启动子的沉默元件中的第二靶标。这两个具体目标是父母补助金的重要扩展。蛋白质的相互作用和功能对于充分了解基因启动子DNA二级结构的生物学功能至关重要，而G4相互作用蛋白的鉴定是实现这一目标的第一步。通过纳入富c链中形成的i基序，该修订将把亲本的范围扩大到c-Myc启动子中沉默元件的完整分子系统，以用于药物靶向。我们的长期目标是使用基于结构的方法来合理设计特异性结合DNA二级结构和调节基因转录的小分子化合物。修订申请将明显加快针对基因启动子区域形成的DNA二级结构的研究，用于抗癌药物的开发。这两个具体目标不是高度相互依赖的，都很有可能在两年内完成。

项目成果

The major G-quadruplex formed in the human BCL-2 proximal promoter adopts a parallel structure with a 13-nt loop in K+ solution.

• DOI: 10.1021/ja4118945
• 发表时间: 2014-02-05
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Agrawal P;Lin C;Mathad RI;Carver M;Yang D
• 通讯作者: Yang D

The dynamic character of the BCL2 promoter i-motif provides a mechanism for modulation of gene expression by compounds that bind selectively to the alternative DNA hairpin structure.

• DOI: 10.1021/ja410934b
• 发表时间: 2014-03-19
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Kendrick S;Kang HJ;Alam MP;Madathil MM;Agrawal P;Gokhale V;Yang D;Hecht SM;Hurley LH
• 通讯作者: Hurley LH

Solution structure of the major G-quadruplex formed in the human VEGF promoter in K+: insights into loop interactions of the parallel G-quadruplexes.

• DOI: 10.1093/nar/gkt784
• 发表时间: 2013-12
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Agrawal P;Hatzakis E;Guo K;Carver M;Yang D
• 通讯作者: Yang D