Development of a Zebrafish PD Drug Discovery Platform

斑马鱼 PD 药物发现平台的开发

• 批准号: 9372289
• 负责人: HOWARD I SIROTKIN
• 金额: $ 7.7万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9372289
• 关键词: Acoustics; Alleles; Animal Model; Behavior; Behavioral; Behavioral Assay; Biological Assay; Brain imaging; Chemicals; Clinical; Clinical Trials; Development; Disease; Disease model; Engineering; Environmental Risk Factor; Etiology; Frequencies; Functional disorder; Gene Targeting; Genetic; Goals; Human; Immunohistochemistry; Invertebrates; LRRK2 gene; Larva; Lesion; Leucine-Rich Repeat; Light; Link; Methods; Midbrain structure; Modeling; Molecular; Movement; Musculoskeletal Equilibrium; Mutation; Neurodegenerative Disorders; Organism; Parkinson Disease; Parkinsonian Disorders; Patients; Phenotype; Phosphotransferases; Physiological; Population; Preclinical Drug Evaluation; Process; Reporter; Research; Resolution; Rodent Model; Speed; Stimulus; Substantia nigra structure; Testing; Therapeutic; Therapeutic Agents; Transgenic Organisms; United States; Validation; Water; Zebrafish; base; dopaminergic neuron; drug discovery; experimental study; flexibility; gain of function; high throughput screening; kinase inhibitor; motor impairment; mutant; nervous system disorder; neuron loss; response; small molecule; stem

It is imperative that we develop rational and systematic approaches to drug discovery to treat Parkinson's and other neurological disorders. Rodent models are essential to this process, but they may not be the optimal starting place. The LRRK2 G2019S allele is the most common Parkinson's disease mutation and corresponding animal models have been developed in a variety of organisms. While the existing models are valuable, we engineered a zebrafish LRRK2 G2019S line that is a uniquely powerful starting place to accelerate PD therapeutics. The experimental flexibility of zebrafish including rapid gene targeting and unparalleled high-resolution functional brain imaging empowers studies of molecular, cellular and physiological mechanism of disease. However, the true power of zebrafish to promote PD research lies in the ability to conduct transformative experiments that cannot readily be done with other models. The ability to culture larvae in 96-well dishes and introduce test compounds via the water enables high throughput screening of small molecules using behavioral readouts. Tens of thousands of compounds can be assayed to identify potential therapeutic agents that can then be further refined to streamline the drug discovery pipeline. Our long-term goal is to perform such a screen to search for compounds that modulate Parkinsonian related behaviors using a zebrafish PD model. In order for this approach to be effective, behaviors that reflect those in patients must first be demonstrated in our LRRK2 G2019S mutant. The objectives of this study are to assay PD-related behaviors and dopaminergic populations in the zebrafish LRRK2 G2019S mutants to determine whether this model is a suitable substrate for high throughput chemical screens. If successful, this approach would go beyond identification of direct LRRK2 inhibitors and enable isolation of compounds that influence other processes to compensate for LRRK2 dysfunction.

我们必须制定合理和系统的药物治疗方法， 治疗帕金森氏症和其他神经系统疾病的新发现。啮齿动物模型是 这对这一进程至关重要，但它们可能不是最佳起点。LRRK2 G2019S等位基因是最常见的帕金森病突变， 已经在多种生物体中开发了动物模型。而现有的 模型是有价值的，我们设计了一个斑马鱼LRRK2 G2019S线，这是一个独特的 这是加速PD治疗的有力起点。 斑马鱼的实验灵活性，包括快速基因打靶和 无与伦比的高分辨率功能性脑成像使分子研究成为可能， 疾病的细胞和生理机制。然而，斑马鱼的真正力量 促进PD研究的关键在于进行变革性实验的能力， 这是其他型号无法做到的。在96孔培养皿中培养幼虫的能力 并且通过水引入测试化合物使得能够高通量筛选小的 使用行为读数来分析分子。可以分析成千上万种化合物 以确定潜在的治疗剂，然后可以进一步完善，以简化 药物研发管道我们的长期目标是执行这样的屏幕搜索 使用斑马鱼PD模型调节帕金森病相关行为的化合物。 为了使这种方法有效，反映患者行为的行为必须 首先在我们的LRRK2 G2019S突变体中得到证实。本研究的目的是 测定斑马鱼LRRK2中的PD相关行为和多巴胺能群体 G2019S突变体，以确定该模型是否是高表达的合适底物。 通过化学筛选。如果成功，这种方法将超越 直接LRRK2抑制剂的鉴定和能够分离 影响其他过程以补偿LRRK2功能障碍。