The Alzheimer's Therapeutics Screening Assay: a high-throughput drug-discovery platform utilizing neurons and microglia derived from human induced pluripotent stem cells and Kinetic Image Cytometry

阿尔茨海默病治疗筛选试验：利用源自人类诱导多能干细胞的神经元和小胶质细胞和动态图像细胞计数的高通量药物发现平台

• 批准号: 9681359
• 负责人: PATRICK M MCDONOUGH
• 金额: $ 30.11万
• 依托单位: VALA SCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9681359
• 关键词: Acetylcholinesterase Inhibitors; Adult; Age; Alleles; Alzheimer' s Disease; Alzheimer' s disease risk; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Antiepileptic Agents; Apolipoprotein E; Apoptosis; Area; Astrocytes; Binding; Biological Assay; Biological Markers; Biological Models; Biomedical Engineering; Brain; Calcium; Calcium Channel Blockers; Catalytic Domain; Cell Cycle Kinetics; Cell Death; Cell surface; Cells; Cellular Morphology; Cellular Stress; Clinical Trials; Coculture Techniques; Cognition; Computer software; Dementia; Development; Drug Modelings; Embryo; Exhibits; FDA approved; Family; Family history of; Gender; Genes; Genetic Risk; Glutamates; Goals; Hippocampus (Brain); Homozygote; Human; Hyperactive behavior; Image Analysis; Image Cytometry; Immune; Impaired cognition; Inflammatory; Inherited; Kinetics; Late Onset Alzheimer Disease; Libraries; Link; Magnetic Resonance; Memantine; Memory Loss; Methods; Microglia; Microscope; Microscopy; Monitor; Mus; Mutation; NMDA receptor antagonist; Neurites; Neurodegenerative Disorders; Neurons; Optical Methods; Pathologic; Pathology; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Pre-Clinical Model; Presenile Alzheimer Dementia; Process; Protein Isoforms; Proteins; Punch Biopsy; Reducing Agents; Research; Research Contracts; Research Personnel; Risk Factors; Rodent; Running; Science; Senile Plaques; Skin; Stem cells; Stimulus; Stress; Structure; Symptoms; Synapses; System; TREM2 gene; Testing; Therapeutic; Toxic effect; abeta oligomer; amnestic mild cognitive impairment; apolipoprotein E-3; apolipoprotein E-4; brain cell; cell type; cellular imaging; cytokine; design; digital; drug development; drug discovery; excitotoxicity; exhaustion; functional MRI scan; gamma secretase; genetic variant; human stem cells; indexing; induced pluripotent stem cell; mild cognitive impairment; movie; neuron loss; neurotoxic; novel strategies; novel therapeutics; presenilin-1; prevent; receptor; screening; specific biomarkers; tau aggregation; voltage

Alzheimer’s Disease (AD), the most common cause of dementia, increases with age, afflicts 5.5 million people in the US, and there is no cure. This phase I project will initiate development of the Alzheimer’s Therapeutics Screening Assay (ATSA), which will utilize neurons, microglia, and astrocytes, derived from human induced pluripotent stem cells. A new testing system is desperately needed as previously FDA-approved therapeutics for AD have limited and temporary effects, and no new drug has been approved for AD since 2003; the many failed clinical trials since then illustrate that current preclinical models are not sufficiently predictive for AD. As amyloid beta peptide (A) plaques accumulate in the brains of AD patients, pharmaceutical companies have exhaustively targeted the beta-amyloid cascade pathway for drug-discovery. However, while many of these agents reduced plaques, none effectively slowed or prevented memory loss in clinical trials. Functional magnetic resonance (fMRI) studies show that subjects with amnestic mild cognitive impairment (aMCI), a prodromal symptom of AD, display hyperactive hippocampal neurons prior to plaque formation, suggesting that dysregulation of neuronal calcium may underlie AD. Relatedly, microglia, the resident immune cells of the brain, are activated in AD and secrete glutamate and inflammatory cytokines that may alter neuronal calcium. Furthermore, the greatest genetic risk for AD is isoform 4 of apolipoprotein E (APOE4), and proteolytic fragments of APOE4 cause dysregulation of intracellular calcium and are toxic to neurons and microglia. For the ATSA, methods will be developed in which hiPSC-neurons, -MG, and -astrocytes are cocultured and AD- relevant stresses (e.g., APOE4 fragments or A oligomers) added, and the cells monitored for alterations in calcium (or voltage) using Kinetic Image Cytometry, a digital microscopy method that quantifies calcium transient activity on a cell-by-cell basis. Cell morphology (neurites, synaptic puncta) and relevant biomarkers will also be quantified. The ATSA will be versatile, and phase II goals will include utilizing iPSCs from patients with inherited or sporadic AD, representing both genders, will be incorporated to further interrogate AD-relevant pathways. Once developed, the ATSA platform will enable testing of hundreds of compounds per day to identify those with beneficial effects against AD related stresses. The ATSA will be marketed by Vala Sciences Inc for contract research to pharmaceutical companies developing novel therapeutics for AD, and used for internal drug-discovery by Vala researchers.

阿尔茨海默氏病（AD）是痴呆症的最常见原因，随着年龄的增长而增加，折磨了550万人 在美国，没有治愈方法。该阶段I项目将启动阿尔茨海默氏症治疗剂的开发 筛选分析（ATSA），该测定法将利用人类诱导的神经元，小胶质细胞和星形胶质细胞 多能干细胞。迫切需要一种新的测试系统，因为以前是FDA批准的疗法 因为AD具有有限和暂时的影响，自2003年以来，没有批准AD的新药；许多人 此后的临床试验失败说明，当前的临床前模型对AD的预测不足。作为 淀粉样蛋白β胡椒（A）斑块积累在广告患者的大脑中，制药公司有 详尽地针对β-淀粉样蛋白级联级联途径进行药物发现。但是，其中许多 药物减少了斑块，在临床试验中没有有效减慢或阻止记忆力丧失。功能 磁共振（fMRI）研究表明，具有柔滑的轻度认知障碍的受试者（AMCI），A AD的前驱症状，在形成牙菌斑之前显示多活跃的海马神经元，表明 神经元钙的失调可能是AD的基础。相关的小胶质细胞，大脑的常驻免疫球， 在AD和秘密谷氨酸和可能改变神经元钙的炎性细胞因子中被激活。 此外，AD的最大遗传风险是载脂蛋白E（APOE4）和蛋白水解的同工型4 APOE4的片段会导致细胞内钙的失调，对神经元和小胶质细胞有毒。为了 将开发ATSA，其中hipsc-神经元，-mg和 - 胃细胞是共培养的，并且 添加了相关应力（例如APOE4片段或A低聚物），并且细胞监测了改变的改变 钙（或电压）使用动力学图像细胞仪，一种数字显微镜方法，可量化钙 逐个细胞基础瞬态活性。细胞形态（神经突，突触点）和相关的生物标志物 也将被量化。 ATSA将具有多功能性，第二阶段目标将包括使用患者的IPSC 将纳入代表这两个性别的遗传或零星广告，以进一步询问与广告相关的 途径。一旦开发，ATSA平台将每天对数百种化合物进行测试 确定针对AD相关应力的有益作用的人。 ATSA将由Vala Sciences销售 INC向制造AD的新疗法的制药公司合同研究，并用于 瓦拉研究人员的内部药物发现。