Small molecule modulators for mitochondrial protein import

用于线粒体蛋白质输入的小分子调节剂

• 批准号: 7694186
• 负责人: Carla M Koehler
• 金额: $ 2.5万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7694186
• 关键词: Affect; Animal Model; Apoptosis; Biogenesis; Biological Assay; Biological Factors; Biological Models; Cell model; Cells; Chemicals; Chemistry; Collaborations; Collection; Communities; Complex; Consult; Cultured Cells; Data; Defect; Degenerative Disorder; Development; Disease; Event; Funding; Goals; Growth; High temperature of physical object; Individual; Inherited; Ischemia; Lead; Libraries; Link; Malignant Neoplasms; Mammals; Medical; Membrane; Membrane Proteins; Metabolism; Mitochondria; Mitochondrial Diseases; Mitochondrial Proteins; Modeling; Mohr-Tranebjaerg syndrome; Molecular; Molecular Bank; Mus; Muscle; Mutation; Myocardial Infarction; Myopathy; Nerve Degeneration; Neurodegenerative Disorders; Pathway interactions; Play; Production; Protein Import; Proteins; Public Health; Research; Role; Saccharomyces cerevisiae; Screening procedure; Signal Transduction; Specificity; Stroke; Structure; Structure-Activity Relationship; System; Temperature; Testing; Therapeutic; Therapeutic Agents; Translating; Vertebrates; Yeasts; Zebrafish; analog; base; chemical genetics; design; dystonia-deafness syndrome; high throughput screening; in vivo; inhibitor/antagonist; mitochondrial dysfunction; mouse model; mutant; novel; novel therapeutics; public health relevance; relating to nervous system; small molecule; small molecule libraries; success; temperature sensitive mutant; therapeutic development; tool; translocase

DESCRIPTION (provided by applicant): Defects in mitochondrial biogenesis lead to a broad range of diseases including neurodegeneration, stroke, myocardial infarction, ischemia, and cancer; however, therapies to correct such diseases are not readily available. We propose to conduct a high throughput screen in the Molecular Libraries Production Center Network (MLPCN) identical to the one that, on a smaller screening scale, has already successfully identified inhibitors of the mitochondrial TIM22 protein import pathway that is required for the assembly of inner membrane proteins in the model organism Saccharomyces cerevisiae. Specifically, we have devised a growth-based assay using a yeast temperature-sensitive mutant that is compromised for growth at a high temperature because of a defective import system. In the screen, we select for small molecules that are synthetically lethal with the mutant strain at a temperature that normally permits growth. Using this assay, we have identified small molecules that specifically target the TIM22 import pathway in yeast mitochondria. The aims of this proposal are to (1) identify small molecules that target the TIM22 import pathway and alter its function and then develop analogs for structure activity relationship (SAR) studies to identify specific chemical compounds that modulate this pathway and (2) utilize these tools in secondary assays and develop probes that we can translate to vertebrate systems to probe mitochondrial function, including the link to mitochondrial diseases, because protein import is highly conserved from yeast to mammals. These studies will result in validated chemical probes for mechanistic studies of mitochondrial import and for potentially inducing/abrogating mitochondrial diseases. A defective TIM22 import pathway leads to the inherited disease, deafness-dystonia syndrome, which results in neurodegeneration. Given our success, we are confident that many novel compounds will be identified that are pertinent for understanding mitochondrial assembly in vertebrates and potentially serving as tools to characterize the molecular basis of deafness-dystonia syndrome. Generally, the medical importance of events regulated by mitochondrial assembly, such as apoptosis, indicates that the chemical genetic approach may also lead to the identification and development of novel therapeutic agents for diseases affected by dysfunctional mitochondria. Identification of these novel compounds, tied with our expertise in finding targets and our ability to exploit them to more fully understand mechanism, justifies our request to expand this screen through the MLPCN. This study is relevant to public health because it may lead to the development of new therapeutics for degenerative muscular and neural diseases. PUBLIC HEALTH RELEVANCE: This project will develop small molecules as probes to investigate the cause of neurodegenerative and degenerative muscular diseases that are initiated by defects in mitochondrial function, using yeast as a model system because protein import pathways are highly conserved from yeast to mammals. The mitochondrion generates energy for the cell and is linked to a broad range of diseases, including cancer and degenerative muscular and neural diseases. Long-term, this project may lead to the development of therapeutics that modulates mitochondrial function in these diseases.

描述（由申请人提供）：线粒体生物发生的缺陷会导致多种疾病，包括神经变性、中风、心肌梗死、缺血和癌症；然而，纠正此类疾病的疗法并不容易获得。我们建议在分子文库生产中心网络 (MLPCN) 中进行高通量筛选，该筛选与在较小的筛选规模上已成功鉴定线粒体 TIM22 蛋白输入途径的抑制剂相同，该途径是组装内部细胞所需的。模型生物酿酒酵母中的膜蛋白。具体来说，我们设计了一种基于生长的测定法，使用酵母温度敏感突变体，由于导入系统有缺陷，该突变体在高温下生长受到影响。在筛选中，我们选择在通常允许生长的温度下对突变菌株具有合成致死作用的小分子。通过该测定，我们鉴定出了专门针对酵母线粒体中 TIM22 输入途径的小分子。该提案的目的是 (1) 识别靶向 TIM22 输入途径并改变其功能的小分子，然后开发用于结构活性关系 (SAR) 研究的类似物，以确定调节该途径的特定化合物，以及 (2) 利用这些化合物二次分析中的工具并开发探针，我们可以将其转化为脊椎动物系统以探测线粒体功能，包括与线粒体疾病的联系，因为蛋白质输入从酵母到哺乳动物是高度保守的。这些研究将产生经过验证的化学探针，用于线粒体输入的机制研究和潜在诱导/消除线粒体疾病。 TIM22 输入通路缺陷会导致遗传性疾病、耳聋肌张力障碍综合征，从而导致神经退行性变。鉴于我们的成功，我们相信将鉴定出许多新化合物，这些化合物与了解脊椎动物线粒体组装相关，并有可能作为表征耳聋肌张力障碍综合征分子基础的工具。一般来说，由线粒体组装调节的事件（例如细胞凋亡）的医学重要性表明，化学遗传学方法也可能导致针对受线粒体功能障碍影响的疾病的新型治疗剂的鉴定和开发。这些新型化合物的鉴定，与我们在寻找目标方面的专业知识以及我们利用它们来更全面地理解机制的能力相结合，证明了我们通过 MLPCN 扩展这一屏幕的请求是合理的。这项研究与公共卫生相关，因为它可能会导致退行性肌肉和神经疾病新疗法的开发。 公共健康相关性：该项目将开发小分子作为探针，以研究由线粒体功能缺陷引发的神经退行性和退行性肌肉疾病的原因，使用酵母作为模型系统，因为从酵母到哺乳动物的蛋白质输入途径高度保守。线粒体为细胞产生能量，并与多种疾病有关，包括癌症以及退行性肌肉和神经疾病。从长远来看，该项目可能会导致调节这些疾病中线粒体功能的疗法的开发。