Screening lipid modifying enzymes to ameliorate A-beta triggered synaptic loss

筛选脂质修饰酶以改善 A-β 引发的突触损失

基本信息

批准号：
8874327
负责人：
Laura Beth Johnson McIntire
金额：
$ 20万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8874327
关键词：
1-Phosphatidylinositol 4-Kinase Affect Aftercare Alzheimer&apos s Disease Amyloid beta-Protein Amyloid beta-Protein Precursor Animal Model Antibodies Behavioral Benchmarking Biological Assay Brain Catabolism Cell Survival Cell physiology Central Nervous System Diseases Clinical Trials Cognitive deficits Critical Pathways Custom Cyclic AMP-Responsive DNA-Binding Protein Defect Dendritic Spines Detection Development Disease Disease model Down Syndrome Enzymes Family Functional disorder Future Genes Genetic Health In Vitro Investigation Libraries Lipase Lipids Maintenance Mediator of activation protein Memory Mental Retardation Metabolic Pathway Metabolism Methods Modeling Mus Mutation Neurons Pathology Pathway interactions Patients Phenotype Phosphatidylinositols Phospholipase Phospholipase C Phospholipids Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Process Protein Kinase Proteins RNA Interference Reporting Research Resistance Resources Risk Role Secondary to Signal Transduction Staging Stem cells Synapses System Testing Therapeutic Toxic effect Transformed Cell Line Validation Viral Virus Diseases Work aging population amyloid peptide base cell type cellular targeting clinical investigation density disease phenotype drug discovery embryonic stem cell enzyme activity global health hippocampal pyramidal neuron immunoreactivity in vivo Model knock-down loss of function miniaturize mouse model neurotransmission novel particle peptide A pre-clinical prevent prophylactic response screening small hairpin RNA synaptic function synaptojanin therapeutic target trafficking transcription factor

项目摘要

DESCRIPTION (provided by applicant): Alzheimer's disease is predicted to be one of the highest priority global health risks in coming decades, yet there are currently no effective prophylactic or disease modifying therapies. The primary focus in the field has been to target reduction of the synaptotoxic amyloid �-peptide (A�) however, recent late-stage clinical trials have proven disappointing. Therefore, new strategies and cellular targets are required. Recent work has found that genetic alteration of lipid modifying enzymes can ameliorate behavioral and synaptic defects in mouse models of the disease. Phospholipases, lipid kinases, lipid activated protein kinases and a lipid phosphatase have all effectively been targeted for ameliorating behavioral deficits in mouse models of the disease indicating that phospholipid modulation may be a key factor of the disease phenotype. A specific class of phospholipids, phosphoinositides (PI), known to be critical for cell and neuronal signaling, has been shown historically and in recent work to be altered in AD affected patient brain as well as in mouse models. The activities of enzymes that control phosphoinositide metabolism and phosphorylation are extensively interconnected and represent a family of tractable lipid modifying enzymes which control levels of signaling lipids. Specifically, the critical signaling lipid phosphatidylinositol-4,5-bisphosphae [P(4,5)P2], is altered by treatment of neurons with A� and in synapses of mouse models of the disease. In order to study lipid changes in response to A�, a physiologically relevant neuronal model is required. Mouse embryonic stem cell derived neurons (ESN) are a scalable neuronal model amenable to mid to high throughput platforms and can be used effectively for screening paradigms. Use of ESN overcomes the limitations of other transformed cell lines commonly used for screening by expression of critical neuronal phenotypes as well as the limitation of dissociated neuronal cultures by scalability. ESN form morphologically mature and functional synapses which are sensitive to A� challenge resulting in a loss of synaptic connections. These neurons display a prominent loss of post-synaptic protein 95 (PDS-95) after treatment with A�. We propose to establish and miniaturize an assay to detect synapse/PSD-95 loss and use RNA interference to screen candidate components of the PI metabolic pathway for synapse maintenance. Since previous research has identified several PI modifying enzymes in connection with AD phenotypes, we expect that this system-wide approach will be able to identify both known and novel targets for amelioration of A�-triggered synaptic loss as well as validate the high content screening platform. These studies will support future investigations into the role of PI in cellular pathways underlying A�-triggered deficits as well as pave the way for more extensive screening paradigms using ESN.

描述（由适用提供）：预计阿尔茨海默氏病被预计是未来几十年中最高的全球健康风险之一，但目前尚无有效的预防性或疾病修改疗法。该领域的主要重点是靶向减少突触毒素淀粉样蛋白肽（A），但是，最近的后期临床试验被证明令人失望。因此，需要新的策略和细胞目标。最近的工作发现，脂质修饰酶的遗传改变可以改善疾病小鼠模型中的行为和突触缺陷。磷脂酶，脂质激酶，脂质激活的蛋白激酶和脂质磷酸酶均有效地靶向改善该疾病的小鼠模型中，这表明磷脂调节可能是疾病表型的关键因素。一类特定的磷脂，磷酸肌醇（PI），已知对于细胞和神经元信号至关重要，已经显示出历史上的历史和最近的工作，在AD受影响的患者脑以及小鼠模型中都会改变。控制磷酸肌醇代谢和磷酸化的酶的活性被广泛互连，并代表了控制信号脂质水平的可拖动脂质修饰酶的家族。具体而言，临界信号传导脂质磷脂酰肌醇-4,5-双磷酸[P（4,5）P2]通过使用A的神经元和疾病小鼠模型突触中的神经元处理来改变。为了研究脂质的变化，需要一个与物理相关的神经元模型。小鼠胚胎干细胞衍生的神经元（ESN）是可扩展的神经元模型，可适应于中部至高吞吐量平台，可有效地用于筛选范例。 ESN的使用克服了通过表达临界神经元表型的表达以及通过可扩展性通过临界神经元表型的表达以及解离神经元培养的局限性来克服其他转化的细胞系的局限性。 ESN形成了形态成熟和功能突触，这些突触对挑战敏感，导致突触连接的丧失。这些神经元在用A的处理后表现出突触后蛋白95（PDS-95）的明显丧失。我们建议建立和微型化测定法以检测突触/PSD-95损失，并使用RNA干扰来筛选PI代谢途径的候选组件以维持突触。由于先前的研究已经确定了与AD表型相关的几种PI修饰酶，因此我们希望这种全系统的方法能够确定已知和新的目标，以改善A触发的合成损失以及验证高含量筛选平台。这些研究将支持未来的调查 PI在A触发的基础细胞途径中的作用定义并为使用ESN更广泛的筛选范式铺平了道路。