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导致广泛的疾病，包括神经变性，中风，心肌梗塞，缺血和癌症；但是，纠正此类疾病的疗法不容易获得。我们建议在分子库生产中心网络（MLPCN）中进行高吞吐量屏幕，该网络与较小的筛选量表相同，该屏幕已经成功地鉴定出了线粒体Tim22蛋白导入途径的抑制剂，这是模型有机体中内膜蛋白所需的线粒体TIM22蛋白进口途径。具体而言，我们已经使用酵母温度敏感的突变体设计了一种基于生长的测定法，该突变体因进口系统缺陷而在高温下被损害。在屏幕上，我们选择在通常允许生长的温度下与突变菌株合成致死的小分子。使用此测定，我们已经确定了专门针对酵母线粒体中TIM22进口途径的小分子。 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从酵母到哺乳动物保守。这些研究将导致验证的化学探针，以进行线粒体进口和潜在诱导/废除线粒体疾病的机理研究。 TIM22进口途径有缺陷导致遗传性疾病，聋哑疾病综合征，导致神经变性。鉴于我们的成功，我们有信心确定许多新型化合物，这些化合物与理解脊椎动物中的线粒体组装有关，并有可能用作表征聋哑疾病的分子基础的工具。通常，线粒体组件调节的事件的医学重要性（例如凋亡）表明，化学遗传学方法还可能导致鉴定和开发受功能障碍线粒体影响的疾病的新型治疗剂。识别这些新颖的化合物，与我们在寻找目标的专业知识以及利用它们以更充分了解机制的能力相关的依据，证明了我们通过MLPCN扩展此屏幕的要求。这项研究与公共卫生有关，因为它可能导致发展新的肌肉和神经疾病的新疗法。 公共卫生相关性：该项目将开发小分子作为探针，以研究神经退行性和退化性肌肉疾病的原因，这些疾病是由线粒体功能中缺陷引发的，使用酵母作为模型系统，因为蛋白质导入途径是从酵母到哺乳动物的高度保守的。线粒体为细胞产生能量，并与广泛的疾病有关，包括癌症和退化性肌肉和神经疾病。长期，该项目可能会导致疗法的发展，从而调节这些疾病中的线粒体功能。