Screening of FoxA1-ER-DNA disruptors for development of breast cancer therapeutic

筛选 FoxA1-ER-DNA 干扰物用于开发乳腺癌治疗药物

基本信息

批准号：
8200699
负责人：
Mary Szatkowski Ozers
金额：
$ 21.82万
依托单位：
PROTEOVISTA, LLC
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-16 至 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8200699
关键词：
Affect Affinity Antineoplastic Agents Area Base Pairing Binding Binding Sites Breast Cancer Cell Cancer Patient Cessation of life DNA DNA Binding DNA Microarray Chip DNA Probes DNA Sequence Data Development Drug Delivery Systems Drug Design Estrogen Receptor alpha Estrogen Receptors Estrogens Factor X Fluorescence G-substrate Gene Expression Regulation Gene Targeting Genes Genetic Transcription Glucocorticoid Receptor Human Genome Incubated Inflammation Left Ligands Maps Marketing Mediating Modeling Molecular Target NR4A1 gene Patients Peptide Nucleic Acids Pharmacologic Substance Phase Physiological Proteins Regulation Resistance Response Elements Screening procedure Site Slide Specificity Tamoxifen Technology Testing Therapeutic Time Woman base design drug discovery genome wide association study high throughput screening improved malignant breast neoplasm new technology novel preference protein complex small molecule stem therapeutic target tool transcription factor

项目摘要

DESCRIPTION (provided by applicant): Estrogen receptor a (ERa) is the major therapeutic target for breast cancer, and ERa binding at target genes in breast cancer cells is guided by DNA binding pioneer factor proteins such as FoxA1. Although tamoxifen is a successful therapeutic for suppressing ER-mediated gene regulation, resistance to tamoxifen occurs in most breast cancer patients within five years and breast cancer remains one of the leading causes of women's death worldwide, highlighting the critical need for improved strategies. With an estimated annual breast cancer therapeutic market of $3-5 billion, the market is receptive to directed strategies. A major challenge in ERa drug design is the necessity to identify small-molecules that specifically target the molecular mechanisms of aberrant gene regulation in breast cancer, while leaving the physiological benefits of ERa action intact. The most critical interaction for ERa-mediated gene transcription occurs at its estrogen response element (ERE) DNA site, and FoxA1 enhances ERa binding at EREs in breast cancer cells. However, DNA-directed therapeutics focused on modulating the ERa-FoxA1-DNA interaction are under- studied areas of drug design because technologies to comprehensively examine this interaction are not yet amenable to high throughput drug discovery. We have developed Cognate Site Identifier (CSI) DNA microarrays, containing every permutation of a 10 to 12 base pair DNA sequence within duplex DNA, and Sequence Specificity Landscapes (SSLs) as a tool to analyze the million-plus data points resulting from the CSI array. This proposal will focus on understanding the comprehensive DNA binding specificity of FoxA1- ERa in breast cancer and specifically disrupting FoxA1 enhancement of ERa binding with DNA-directed peptide nucleic acids (PNAs). The specific aims of this Phase I proposal are: 1. The DNA binding preferences of FoxA1 will be characterized using FoxA1 from breast cancer cell lysates and as a purified protein, by CSI-SSL analysis. These studies will identify direct DNA binding sites of purified FoxA1 as well as potential protein-tethered FoxA1-DNA interactions in breast cancer cells. 2. DNA-binding PNAs will be designed to specifically target and disrupt the FoxA1-DNA interaction, as model therapeutics to down-regulate FoxA1-mediated transcription at genes instrumental in breast cancer regulation. 3. A high throughput screening platform will be developed using CSI DNA microarrays to examine the ERa- FoxA1-DNA interaction and screen test compounds as targeted disruptors of this interaction. Our findings will define a new arena for DNA-based therapeutics by expanding CSI-SSL technology to a DNA-bound heterodimeric protein complex, towards small-molecule screening, discovery of novel drug targets, and development of targeted DNA binding molecules as therapeutics for breast cancer. PUBLIC HEALTH RELEVANCE: Estrogen receptor alpha (ERa) is the major therapeutic target for breast cancer, and ERa binding at target genes in breast cancer cells is guided by DNA binding pioneer factor proteins such as FoxA1. While tamoxifen represents a first-line pharmaceutical defense for ERa-positive breast cancer, the field is in critical need of directed therapies because approximately 30% of patients are initially recalcitrant to tamoxifen and, of those who do respond, virtually all will become resistant to tamoxifen within five years. We are developing new technologies, the Cognate Site Identifier DNA microarray and Sequence Specificity Landscape, as a high throughput platform to rapidly identify compounds that specifically target and disrupt ERa-FoxA1-DNA interactions, for discovery of novel DNA-directed breast cancer drugs.

描述（由申请人提供）：雌激素受体A（ERA）是乳腺癌的主要治疗靶点，而乳腺癌细胞中靶基因的ERA结合则由DNA结合先锋因子蛋白（如FOXA1）指导。尽管他莫昔芬是抑制ER介导的基因调节的成功治疗方法，但在五年内，大多数乳腺癌患者都会发生对他莫昔芬的耐药性，而乳腺癌仍然是全球妇女死亡的主要原因之一，强调了对改善策略的关键需求。该市场估计每年的乳腺癌治疗市场为3-5亿美元，可以接受指示策略。 ERA药物设计中的一个主要挑战是必须确定针对乳腺癌中异常基因调节的分子机制的小分子，同时使ERA动作的生理益处完好无损。 ERA介导的基因转录的最关键相互作用发生在其雌激素反应元件（ERE）DNA位点，FOXA1增强了乳腺癌细胞中ERE的ERA结合。然而，DNA指导的治疗剂侧重于调节ERA-FOXA1-DNA相互作用的药物设计领域，因为可以全面检查这种相互作用的技术尚不适合高通量药物发现。我们已经开发了同源位点标识符（CSI）DNA微阵列，其中包含双链DNA中10至12个基对DNA序列的每一个置换，以及序列特异性景观（SSL）作为分析CSI阵列产生的百万含量数据点的工具。该建议将集中于了解乳腺癌中FOXA1时代的全面DNA结合特异性，并特别破坏FOXA1与DNA指导的肽核酸（PNAS）的ERA结合增强。该阶段提案的具体目的是：1。通过CSI-SSL分析，使用乳腺癌细胞裂解物和纯化蛋白的FOXA1的DNA结合偏好将以FOXA1为特征。这些研究将确定纯化FOXA1的直接DNA结合位点以及乳腺癌细胞中潜在的蛋白质螺旋蛋白FOXA1-DNA相互作用。 2。DNA结合PNA的设计旨在特异性靶向和破坏FOXA1-DNA相互作用，作为模型疗法，以下调FOXA1介导的基因在乳腺癌调节中有助于基因的转录。 3。将使用CSI DNA微阵列开发高吞吐量筛选平台，以检查ERA-FOXA1-DNA相互作用和筛查测试化合物作为这种相互作用的靶向破坏者。我们的发现将通过将CSI-SSL技术扩展到DNA结合的异二聚体蛋白复合物，朝着小分子筛选，发现新型药物靶标以及靶向DNA结合分子作为乳腺癌的治疗剂来定义一个新的基于DNA的治疗疗法的领域。公共卫生相关性：雌激素受体α（ERA）是乳腺癌的主要治疗靶点，而乳腺癌细胞中靶基因的ERA结合由DNA结合先锋因子蛋白（例如FOXA1）指导。他莫昔芬代表了ERA阳性乳腺癌的一线药物防御，但该领域迫切需要有定向疗法，因为大约30％的患者最初是对他莫昔芬的顽固耐药的，并且在五年内，几乎所有反应的患者都会在五年内对他的他莫昔芬有抵抗力。我们正在开发新技术，同源位点标识符DNA微阵列和序列特异性景观，作为一个高吞吐量平台，可快速识别专门针对和破坏ERA-FOXA1-DNA相互作用的化合物，以发现新型DNA乳腺癌药物。