High-throughput Chemotyping of Yeast Signature Strains Reflecting Hsp90 Biology

反映 Hsp90 生物学特征的酵母特征菌株的高通量化学分型

• 批准号: 7425740
• 负责人: Vincent Jo Davisson
• 金额: $ 15.25万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7425740
• 关键词: Agonist; Binding; Biochemical; Biological; Biological Assay; Biological Process; Biology; Cell Wall; Chemicals; Class; Classification; Clinical Trials; Complex; Disease; Dose; Enzymes; Genes; Genetic; Genotype; Growth; Hand; Human; Knowledge; Libraries; Measures; Methods; Molecular Bank; Molecular Chaperones; Mutation; Nature; Organism; Pathway interactions; Permeability; Play; Preclinical Drug Evaluation; Proteins; Purpose; Range; Reproducibility; Role; Screening procedure; Series; Solutions; Staging; Two-Hybrid System Techniques; Universities; Validation; Variant; Yeasts; assay development; base; cytotoxic; density; design; high throughput screening; improved; inhibitor/antagonist; interest; knowledge base; neuroprotection; novel; protein protein interaction; repository; research study; small molecule; small molecule libraries; yeast genetics

DESCRIPTION (provided by applicant): The objective of this proposal is to identify a series of novel small molecule modulators of Hsp90-centered biological networks. Hsp90 is an important oncological target and several compound inhibitors are undergoing clinical trials. A growing interest in exploiting the use of Hsp90 inhibitors for other disease states is emerging including the use of sub-cytotoxic concentrations to increase expression levels of Hsp90 to provide neuroprotection. These inhibitors and others have had an invaluable role in establishing our current knowledge of Hsp90 biology. However, the exact mode of action of Hsp90 inhibitors and their effect on the surrounding cellular networks is still unclear. This problem is not trivial due to the intricate and extensive roles Hsp90 plays in myriad biological processes. Identification of additional pharmacological modulators will have a substantial impact in our ability to modulate Hsp90 pathways. We will exploit the unparalleled knowledge base in yeast genetics and the tractability of the organism to high throughput screening (HTS) to develop an assay capable of identifying chemical modulators, including both antagonists and agonists of Hsp90 pathways. Our genetic-based and multifactorial approach determines growth parameters of a set of yeast strains in the presence of compounds. The growth profiles of these signature strains will aid in identifying compound classes and allow functional linkage with stages of the Hsp90 chaperone cycle and specific subsets of Hsp90 pathways. Solutions to traditional problems with using yeast as a drug-screening platform will be outlined and incorporated in our assay development. The assay we are developing will be in a simple and robust high density 384-well experimental format that will allow screening of a large chemical library matrix such as the Molecular Libraries Small Molecule Repository.

描述（由申请人提供）：本提案的目的是鉴定一系列以 Hsp90 为中心的生物网络的新型小分子调节剂。 Hsp90 是重要的肿瘤靶点，多种复合抑制剂正在进行临床试验。人们对利用 Hsp90 抑制剂治疗其他疾病状态越来越感兴趣，包括使用亚细胞毒性浓度来增加 Hsp90 的表达水平以提供神经保护。这些抑制剂和其他抑制剂在建立我们当前的 Hsp90 生物学知识方面发挥了不可估量的作用。然而，Hsp90抑制剂的确切作用方式及其对周围细胞网络的影响仍不清楚。由于 Hsp90 在无数的生物过程中发挥着复杂而广泛的作用，这个问题并非微不足道。其他药理学调节剂的鉴定将对我们调节 Hsp90 途径的能力产生重大影响。我们将利用无与伦比的知识库 酵母遗传学和生物体易处理性的高通量筛选 (HTS)，以开发能够识别化学调节剂（包括 Hsp90 途径的拮抗剂和激动剂）的检测方法。我们基于遗传的多因素方法确定了一组酵母菌株在化合物存在下的生长参数。这些特征菌株的生长概况将有助于识别化合物类别，并允许与 Hsp90 伴侣循环阶段和 Hsp90 途径的特定子集进行功能联系。将概述使用酵母作为药物筛选平台的传统问题的解决方案，并将其纳入我们的检测开发中。我们正在开发的检测方法将采用简单而强大的高密度 384 孔实验形式，该形式将允许筛选大型化学库矩阵，例如分子库小分子存储库。