C. elegans as a model organism for human disease through the application of phenologs

通过应用酚类化合物将线虫作为人类疾病的模型生物

基本信息

批准号：
10469264
负责人：
James Inglese
金额：
$ 16.44万
依托单位：
NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10469264
关键词：
Animal Model Area Biological Assay Biological Models Caenorhabditis elegans Cell Membrane Permeability Chemicals Collaborations Cyclic Peptides Cytometry Development Disease Equipment Fluorescence Genes Genetic Engineering Goals Health Human Human Pathology Image Infectious Agent Knock-out Label Laboratories Lasers Life Link Measures Mutation National Institute of Diabetes and Digestive and Kidney Diseases National Institute of Neurological Disorders and Stroke Nematoda Nerve Degeneration Neurons Organism Parkinson Disease Pathway interactions Phenotype Phosphoglycerate Mutase Proteins Sorting - Cell Movement Testing Therapeutic Agents Time Tissues United States National Institutes of Health Work base cell type dopaminergic neuron follow-up gene function genetic strain high throughput screening human disease inhibitor/antagonist instrumentation interest membrane activity novel screening

项目摘要

This project has advanced from our Ipglycermides: Novel potent and selective inhibitors of parasitic phosphoglycerate mutase project where C. elegans serve as a model organism for infectious nematode species to test the activity of membrane permeable cyclic peptides on the viability of the organism. The ADST laboratory has developed a 384-well microtiter plate, laser cytometry-based qHTS assay to evaluate anti-nematodal agents. Primary screening is based on an overall decrease in worm number and area as measured by green fluorescence in a strain of worm ubiquitously expressing GFP. The lab is equipment with the necessary instrumentation for follow-up assays involving high-content imaging and life stage sorting to distinguish various modes-of-action for the potential chemical modulators. In collaboration with Dr. D. Sibley (NINDS, NIH) the ADST laboratory has developed a 384-well qHTS laser cytometry-based screening assay for Parkinsons Disease evaluating neurodegeneration of dopaminergic neurons in C. elegans strains containing two different human PD-linked genes. Both strains express GFP in their dopaminergic neurons allowing for traceable fluorescence intensity as a measure of neuron health over time. The ADST lab also has a long-standing collaboration with Dr. A. Golden (NIDDK, NIH) in working to develop novel genetically engineered heterozygous knockout strains of C. elegans for the Ipglycermide project. The Golden lab has provided expertise in the development of novel genetic strains of worms that may be expanded to additional disease targets.

该项目源自我们的 Ipglycermides：寄生磷酸甘油酸变位酶的新型有效和选择性抑制剂项目，其中秀丽隐杆线虫作为传染性线虫物种的模型生物，以测试膜渗透性环肽对生物体活力的活性。 ADST 实验室开发了一种 384 孔微量滴定板、基于激光细胞计数的 qHTS 测定法，用于评估抗线虫药物。初步筛选基于蠕虫数量和面积的总体减少（通过普遍表达 GFP 的蠕虫品系中的绿色荧光测量）。该实验室配备了必要的仪器，用于后续分析，包括高内涵成像和生命阶段分类，以区分潜在化学调节剂的各种作用模式。 ADST 实验室与 D. Sibley 博士（NINDS、NIH）合作，开发了一种基于 384 孔 qHTS 激光细胞术的帕金森病筛查方法，评估含有两种不同人类 PD 相关基因的线虫菌株中多巴胺能神经元的神经变性。两种菌株的多巴胺能神经元均表达 GFP，从而可追踪荧光强度，作为神经元随时间健康状况的衡量标准。 ADST 实验室还与 A. Golden 博士（NIDDK、NIH）长期合作，致力于为 Ipglycermide 项目开发新型基因工程杂合敲除秀丽隐杆线虫菌株。 Golden 实验室在开发新型蠕虫遗传菌株方面提供了专业知识，这些菌株可能会扩展到其他疾病目标。