High-throughput Chemotyping of Yeast Signature Strains Reflecting Hsp90 Biology

描述（由申请人提供）：本提案的目的是确定一系列以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，几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的兴趣越来越大，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目前的Hsp90生物学知识方面发挥了宝贵的作用。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尚不清楚。由于Hsp90在无数生物过程中发挥着复杂而广泛的作用，因此这个问题并非微不足道。鉴定额外的药理学调节剂将对我们调节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

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.