Selective Inhibitors of Human HSF2

人类 HSF2 的选择性抑制剂

• 批准号: 8464285
• 负责人: Alexandre M. Erkine
• 金额: $ 2.92万
• 依托单位: BUTLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8464285
• 关键词: Affect; Animals; Antineoplastic Agents; Apoptosis; Area; Attention; Biological Assay; Cancer Biology; Carbon; Cardiovascular Diseases; Cardiovascular system; Cell Culture Techniques; Cell Survival; Cell model; Cells; Chemicals; Chimeric Proteins; Chromosomal translocation; Code; Collaborations; Collection; Culture Media; Detection; Development; Evolution; Future; Galactose; Genes; Genetic Transcription; Genomics; Glucose; Growth; HSF1; Heat-Shock Proteins 70; Heat-Shock Proteins 90; Heat-Shock Response; Heat-Shock Transcription Factor 2; Immune system; In Vitro; Individual; Inhibitory Concentration 50; Laboratories; Lead; Libraries; Light; Malignant Neoplasms; Masks; Measures; Messenger RNA; Molecular; Molecular Biology; Molecular Chaperones; Molecular Probes; Mutate; Mutation; Myocardial Infarction; Nerve Degeneration; Neurodegenerative Disorders; Nutrient; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Phenotype; Promega; Property; Regulation; Reporter; Research; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Seeds; Shock; Source; Specificity; Structure-Activity Relationship; System; Time; Translations; United States National Institutes of Health; Work; Yeasts; absorption; base; cancer cell; cell growth; chaperone machinery; design; drug development; in vivo; inhibitor/antagonist; luminescence; microbial; oncology; overexpression; palliative; pre-clinical; promoter; protein misfolding; restoration; screening; small molecule; small molecule libraries; tool; tumorigenesis

DESCRIPTION (provided by applicant): Recent scientific developments indicate that the molecular chaperone system is actively involved in cancerogenesis, neurodegeneration and cardiovascular disorders. The master regulator of molecular chaperone expression at the level of transcription is the heat shock factor (HSF). The specific aim of the proposed research is to setup and implement the screening system for affecters of human HSF2. Inhibitors (in case of cancer) or activators (in case of neurodegenerative and cardiovascular disorders) of HSF can be used as important future drug leads in pharmacology. The screening system design is based on the utilization of a new yeast strain created by our laboratory, in which the conditional overexpression of human HSF2 under Gal1 promoter creates a slow growth phenotype. Potential compounds increasing the growth rate of the tester strain will be verified and used as leads for new drug development. The initial growth rate of the tester strain will be easily adjusted for screens of different stringencies by the incorporation of glucose in the growth media containing galactose as a major carbon source nutrient. The detection of cell growth restoration will be done by registering a change in the light absorption of the cell culture and, in addition, y registering luminescence using the commercially available BacTiter-Glo" Microbial Cell Viability Assay (Promega). Compound hits will be evaluated over a range of concentrations around the initial screening and the IC50 will be determined. Hits will be verified additionally by measuring change in the mRNA level of a set of reporter heat shock genes using real-time quantitative RT-PCR for non-heat shock and heat shock conditions in the presence of the identified compound and compared to similar samples from untreated cells. The advantage of this approach is that the true in vivo affecters of hHSF2 will be identified and confirmed.

描述（由申请人提供）：最近的科学发展表明，分子伴侣系统积极参与癌症发生，神经变性和心血管疾病。在转录水平上调控分子伴侣表达的主要因子是热休克因子（HSF）。本研究的具体目的是建立和实施人类HSF2感染者的筛选系统。HSF的抑制剂（在癌症的情况下）或激活剂（在神经退行性疾病和心血管疾病的情况下）可以用作药理学中重要的未来药物先导。筛选系统的设计是基于利用我们实验室创造的一种新的酵母菌株，其中在Gal1启动子下有条件地过表达人HSF2产生缓慢的生长表型。增加试验菌株生长速率的潜在化合物将被验证并用作新药开发的线索。通过将葡萄糖掺入含有半乳糖作为主要碳源营养物的生长培养基中，测试菌株的初始生长速率将很容易调整为不同强度的筛选。细胞生长恢复的检测将通过记录细胞培养物的光吸收变化来完成，此外，使用市售的BacTiter-Glo“微生物细胞活力测定法”（Promega）记录发光。将在初始筛选前后的一系列浓度范围内评估化合物的影响，并确定IC50。在非热休克和存在已鉴定化合物的热休克条件下，通过使用实时定量RT-PCR测量一组报告基因mRNA水平的变化，并将其与来自未处理细胞的类似样品进行比较，进一步验证命中。这种方法的优点是，hHSF2在体内的真正影响因素将被识别和确认。

项目成果

Nucleosome distortion as a possible mechanism of transcription activation domain function.

• DOI: 10.1186/s13072-016-0092-2
• 发表时间: 2016
• 期刊: Epigenetics & chromatin
• 影响因子: 3.9
• 作者: Erkina TY;Erkine AM
• 通讯作者: Erkine AM