Small Molecule Inhibitors of Cilia

纤毛小分子抑制剂

基本信息

批准号：
7303720
负责人：
Wallace Marshall
金额：
$ 2.5万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-06-01 至 2008-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7303720
关键词：
Affect Animal Model Animals Basic Science Behavior Biological Assay Biological Models Biology Brain Bronchiectasis Candidate Disease Gene Cell Culture System Cell Death Cells Chemicals Chlamydomonas Chronic Chronic Sinusitis Cilia Compatible Computer software Cultured Cells Cytoskeleton Defect Development Dimethyl Sulfoxide Disease Drug Delivery Systems Drug effect disorder Drug usage Dynein ATPase Environment Etiology Event Future Gene Expression Gene Targeting Genetic Goals Haploidy Human Human body Hydrocephalus Image Image Analysis Incubators Lead Length Liquid substance Little&apos s Disease Lung Inflammation Mammalian Cell Marshal Measures Mediating Methods Microscopy Microtubules Modeling Movement Mucous body substance Mutation Numbers Obesity Organelles Organism Performance Pharmaceutical Preparations Pharmacological Treatment Phenotype Play Polycystic Kidney Diseases Population Process Proteins Range Reading Reagent Renal function Reproducibility Research Resolution Retina Retinal Degeneration Retinitis Pigmentosa Robotics Role Role playing therapy Score Sensory Staging Standards of Weights and Measures Surface Swimming Symptoms Water Work base body sense cell motility cell type cilium biogenesis day depolymerization drug development follow-up high throughput screening inhibitor/antagonist innovation katanin lipid metabolism mutant prevent research study response small molecule tissue/cell culture tool

Affect Animal Model Animals Basic Science Behavior Biological Assay Biological Models Biology Brain Bronchiectasis Candidate Disease Gene Cell Culture System Cell Death Cells Chemicals Chlamydomonas Chronic Chronic Sinusitis Cilia Compatible Computer software Cultured Cells Cytoskeleton Defect Development Dimethyl Sulfoxide Disease Drug Delivery Systems Drug effect disorder Drug usage Dynein ATPase Environment Etiology Event Future Gene Expression Gene Targeting Genetic Goals Haploidy Human Human body Hydrocephalus Image Image Analysis Incubators Lead Length Liquid substance Little&apos s Disease Lung Inflammation Mammalian Cell Marshal Measures Mediating Methods Microscopy Microtubules Modeling Movement Mucous body substance Mutation Numbers Obesity Organelles Organism Performance Pharmaceutical Preparations Pharmacological Treatment Phenotype Play Polycystic Kidney Diseases Population Process Proteins Range Reading Reagent Renal function Reproducibility Research Resolution Retina Retinal Degeneration Retinitis Pigmentosa Robotics Role Role playing therapy Score Sensory Staging Standards of Weights and Measures Surface Swimming Symptoms Water Work base body sense cell motility cell type cilium biogenesis day depolymerization drug development follow-up high throughput screening inhibitor/antagonist innovation katanin lipid metabolism mutant prevent research study response small molecule tissue/cell culture tool

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项目摘要

DESCRIPTION (provided by applicant): We have developed a simple and reliable assay to identify small molecule inhibitors of cilia assembly and function. Cilia are sensory and motile organelles that play critical roles in development and disease, including polycystic kidney disease, hydrocephalus, syndromic obesity, chronic sinusitis, bronchiectasis, and retinal degeneration. Small molecule modulators would provide critical tools for studying dynamic events in cilia, and might even point the way to development of pharmacological approaches to ciliary diseases, for which no drugs are currently available. Our assay takes advantage of the model organism Chlamydomonas, which uses cilia to swim to the bottom of an assay well. Because the cells are green, this causes a dark green pellet to self-organize in the well. Mutations or pharmacological treatments which block ciliary motility, or which cause cells to have no cilia, prevent this organization. We have developed image-analysis software that can distinguish these two results in a 96-well format. We have verified the performance of this assay and found it has a Z- value of 0.7 with low plate-to-plate variability and high day-to-day reproducibility. The assay can tolerate DMSO up to 1% and is completely compatible with high throughput robotic methods. We anticipate that a screen using this assay may provide a critical step forward in understanding the mechanisms and treatment of ciliary diseases. Cilia are cellular antennas which project from the surface of most cells in the human body, and sense the surrounding environment. These sensory cilia play critical roles in kidney function, and defects in cilia lead to a devastating disease called polycystic kidney disease. Cilia also play roles in fat metabolism and retina function, and cilia defects can result in genetic obesity and retinal degeneration. Cilia also act like oars to move fluid around cells, and defects in cilia movement can lead to a number of disease symptoms including chronic lung inflammation due to poor mucus clearance, and to hydrocephalus (water on the brain). Currently there are no drugs available to treat diseases, and little is known about how cilia assemble or function. We have developed a new assay for identifying chemical compounds that can affect cilia. These will not only provide important tools for basic research on how cilia work, they would also serve as lead compounds for drug development efforts aimed at treating the large number of ciliary diseases (ciliopathies) that are now known.

描述（由申请人提供）：我们开发了一种简单可靠的测定法，以识别纤毛组装和功能的小分子抑制剂。纤毛是感官和动感细胞器，在发育和疾病中起着关键作用，包括多囊性肾脏疾病，脑积水，综合征肥胖，慢性鼻窦炎，支气管张和视网膜退化。小分子调节剂将为研究纤毛的动态事件提供关键的工具，甚至可能指向开发用于睫状体疾病的药理学方法的方法，目前没有药物可用。我们的测定方法利用了模型有机体衣原体，该衣原体利用纤毛良好地游泳到底部。由于细胞是绿色的，这会导致深绿色的颗粒在井中自组织。阻断睫状运动或导致细胞没有纤毛的突变或药理治疗，阻止了该组织。我们开发了图像分析软件，可以以96孔格式区分这两个结果。我们已经验证了该测定法的性能，并发现它的Z值为0.7，板对板的可变性低和日常可重复性。该测定最大可耐受DMSO，并且与高吞吐量机器人方法完全兼容。我们预计，使用此测定法的屏幕可能会在理解睫毛疾病的机制和治疗方面迈出关键的一步。纤毛是从人体大多数细胞表面投射的细胞天线，并感知周围环境。这些感觉纤毛在肾功能中起着关键作用，纤毛缺陷导致一种毁灭性疾病，称为多囊性肾脏疾病。纤毛在脂肪代谢和视网膜功能中也起着作用，纤毛缺陷会导致遗传肥胖和视网膜变性。纤毛也像桨一样，在细胞周围移动液体，而纤毛运动的缺陷会导致许多疾病症状，包括由于粘液清除率差而引起的慢性肺部炎症和脑积水（大脑的水）。当前，没有可用于治疗疾病的药物，而对于纤毛纤毛的组装或功能知之甚少。我们开发了一种新的测定方法，用于鉴定可能影响纤毛的化合物。这些不仅将提供有关纤毛如何工作的基础研究的重要工具，还将充当旨在治疗现在已知的大量纤毛疾病（纤毛疾病）的药物开发工作的铅化合物。