Patient iPSC derived neural synaptic screen for drug discovery

用于药物发现的患者 iPSC 衍生神经突触筛选

基本信息

批准号：
9141432
负责人：
Wei Zhang
金额：
$ 22.48万
依托单位：
JUVOBIO PHARMACEUTICALS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9141432
关键词：
Address Affect Alzheimer&apos s Disease Animals Architecture Autistic Disorder Behavioral Biological Assay Cell Line Cell model Clinical Cognitive deficits Complex DISC1 gene Defect Development Disease Disease model Etiology Frameshift Mutation Funding Genetic Genetic Risk Glutamates Human Impaired cognition Industry Interneurons Licensing Marketing Medical Mental Depression Mental disorders Modeling Mus Mutant Strains Mice Mutation Neurons Parkinson Disease Patients Pharmaceutical Preparations Pharmacologic Substance Phase Phenotype Population Property Prosencephalon Protocols documentation Public Health Reporting Rett Syndrome Rolipram Schizophrenia Stem cells Susceptibility Gene Symptoms Synapses Therapeutic Therapeutic Effect Translating Work base density disease phenotype drug discovery genetic risk factor induced pluripotent stem cell innovation microdeletion mouse model nerve stem cell novel partial response public health relevance relating to nervous system response social tool treatment effect

项目摘要

DESCRIPTION (provided by applicant): We propose to 1). Use a set of human schizophrenia (SCZ) disease models utilizing induced pluripotent stem cells (iPSCs) derived from SCZ patients to evaluate six advanced SCZ compounds from a pharma partner, to assess their potential to be further developed into SCZ therapeutics; and 2). Discover new compound leads for SCZ. We will use one of the models - a validated synaptic density screen in 384-well format, using forebrain glutamatergic neurons (FGNs) derived from iPSCs bearing a Disrupted in schizophrenia 1 (DISC1) frameshift mutation. We will characterize the hits in the additional SCZ iPSC models. SCZ affects around 1% of the world's population and is a major public health problem. About 30% of SCZ patients do not response to available medications. 30% only show partial response. The industry's hunt for better therapeutics have been thwarted by inadequacy of current drug discovery models. Patient-specific iPSC-derived neurons hold great promises as effective disease models for mechanism studies and drug discovery. Our team has generated and characterized SCZ iPSC lines from patients with different genetic risk factors and background. These cell lines, isogenic and normal controls will be utilized for this proposal. We have optimized differentiation protocols that produce highly pure populations of cortical neural progenitors (NPCs), GABAergic interneurons and FGNs (>99%, >80% and >90% respectively). The purity of differentiated populations allows us to confidently attribute assay readouts to intrinsic properties of the cellular models and experimental manipulations. The high-throughput synaptic screen is validated: 1). Correcting DISC1 mutation in patient iPSCs corrected the synaptic and other functional defects; 2). Introducing DISC1 mutation into normal iPSCs or mice recreated cellular deficits and created animal behavioral and cognitive deficits; 3). A mechanism relevant compound rescues deficits in both DISC1 FGNs and DISC1 mutant mice; 4). Nine other compounds mechanistically related to this compound was shown to rescue synaptic deficit in a separate targeted screen. Aim 1. Evaluate six advanced discovery/development SCZ compounds from a pharma partner for potential continued development for SCZ. We will differentiate 14 SCZ and control iPSC lines into NPCs and then onto FGNs and GABAergic neurons. Treat the neurons for 2-4 weeks with the six compounds at four concentrations. Assess treatment effects with synaptic screen and other morphological and functional assays. Aim 2. Conduct primary screen of 50,000 compounds with synaptic screen and confirmation screens with additional SCZ iPSC models. We will use synaptic density assay with one DISC1 line for primary screen. We will use its isogenic control and another DISC1 line (with different clinical presentation) for counter and confirmation screens, and further characterize with other SCZ lines to generate leads.

描述（由适用提供）：我们建议1）。使用一组利用源自SCZ患者的诱导多能干细胞（IPSC）的人类精神分裂症（SCZ）疾病模型来评估制药伴侣的六种高级SCZ化合物，以评估其进一步发展为SCZ疗法的潜力；和2）。发现SCZ的新化合物线索。我们将使用一种模型之一 - 使用前脑谷氨酸能神经元（FGNS）衍生自IPSC的前脑谷氨酸能神经元（FGNS），以384孔格式进行了经过验证的突触密度屏幕，该筛选源自具有精神分裂症1（盘子1）的frameshift frameshift突变的IPSC。我们将表征其他SCZ IPSC模型中的命中。 SCZ影响了世界人口的1％，这是一个主要的公共卫生问题。大约30％的SCZ患者对可用药物不反应。 30％仅显示部分响应。该行业寻求更好的治疗学，我们已经优化了分化方案，这些方案产生了高度纯净的皮质神经元祖细胞（NPC），GABA能中间神经元和FGN（分别> 99％，> 80％和> 90％）。分化群体的纯度使我们能够将读数归因于细胞模型和实验操作的内在特性。高通量突触屏幕已验证：1）。纠正患者IPSC中的椎间盘1突变纠正了突触和其他功能缺陷； 2）。将圆盘1突变引入正常的IPSC或小鼠中，重现了细胞缺陷，并造成了动物的行为和认知缺陷； 3）。相关的复合机制可挽救盘子1 FGN和盘状突变小鼠的缺陷； 4）。与该化合物机械相关的其他九种化合物被证明可以在单独的靶向屏幕中挽救突触缺乏。 AIM 1。评估来自药品合作伙伴的六种高级发现/开发SCZ化合物，以供SCZ继续开发。我们将区分14个SCZ，并将IPSC线控制为NPC，然后将IPSC线与FGN和GABA能神经元区分开。用六个化合物以四个浓度治疗神经元2-4周。通过合成筛查以及其他形态学和功能测定法评估治疗效果。 AIM 2。使用其他SCZ IPSC型号的合成屏幕和确认屏幕进行50,000种化合物的主要屏幕。我们将使用一条Disc1系列使用突触密度评估进行主屏幕。我们将使用其同源控制和另一个Disc1系（带有不同的临床表现）进行计数器和确认筛选，并进一步以其他SCZ系列来产生潜在客户。