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(申请人提供)：线粒体生物发生缺陷会导致广泛的疾病，包括神经变性、中风、心肌梗死、缺血和癌症；然而，纠正这些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，与已经在较小规模上成功识别模式生物酿酒酵母中组装内膜蛋白所需的线粒体TIM22蛋白输入途径抑制剂的筛选相同。具体地说，我们设计了一种基于生长的检测方法，使用了一种酵母温度敏感突变体，由于进口系统的缺陷，该突变体在高温下生长受到影响。在屏幕上，我们选择在正常允许生长的温度下对突变菌株具有合成致命性的小分子。使用这项测试，我们已经确定了在酵母线粒体中特异性靶向TIM22输入途径的小分子。该建议的目的是(1)确定针对TIM22导入途径的小分子并改变其功能，然后开发类似物用于结构活性关系(SAR)研究以确定调控这一途径的特定化合物；以及(2)利用这些工具进行二次分析并开发我们可以翻译到脊椎动物系统的探针以探测线粒体功能，包括与线粒体疾病的联系，因为蛋白质从酵母到哺乳动物的导入是高度保守的。这些研究将为线粒体输入的机制研究和潜在地诱导/消除线粒体疾病带来有效的化学探针。TIM22导入途径的缺陷会导致遗传性疾病--耳聋-肌张力障碍综合征，从而导致神经变性。鉴于我们的成功，我们相信将发现许多新的化合物，这些化合物与了解脊椎动物线粒体组装有关，并可能成为表征耳聋-肌张力障碍综合征分子基础的工具。通常，线粒体组装调节的事件，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发现和开发新的治疗药物来治疗受线粒体功能障碍影响的疾病。这些新化合物的鉴定与我们在寻找目标方面的专业知识和我们利用它们以更充分地了解机制的能力相联系，证明我们有理由要求通过MLPCN扩大这一筛查。这项研究与公共卫生有关，因为它可能导致退行性肌肉和神经疾病的新疗法的发展。 与公共卫生相关：该项目将开发小分子作为探针，以调查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是高度保守的。线粒体为细胞产生能量，并与一系列疾病有关，包括癌症和退行性肌肉和神经疾病。从长远来看，这个项目可能会导致对这些疾病中线粒体功能进行调节的治疗方法的发展。