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（由申请人提供）：阿尔茨海默氏病预计将成为未来几十年最优先的全球健康风险之一，但目前尚无有效的预防或疾病缓解疗法。该领域的主要焦点是减少阿尔茨海默氏病。然而，最近的后期临床试验结果令人失望，因此，需要新的策略和细胞靶点，最近的工作发现脂质修饰酶的基因改变可以改善这种情况。该疾病小鼠模型中的行为和突触缺陷磷脂酶、脂质激酶、脂质激活蛋白激酶和脂质磷酸酶都已有效地改善该疾病小鼠模型的行为缺陷，表明磷脂调节可能是该疾病的关键因素。一类特定的磷脂，即磷酸肌醇 (PI)，已知对细胞和神经元信号传导至关重要，历史上和最近的研究都表明它在 AD 中会发生改变。控制磷酸肌醇代谢和磷酸化的酶的活性广泛地相互关联，代表了控制信号脂质水平的易处理的脂质修饰酶家族，特别是关键的信号脂质磷脂酰肌醇-4,5。 -bisphosphae [P(4,5)P2]，通过用 A� 处理神经元和该疾病小鼠模型的突触而改变。响应 A� 的脂质变化是必需的，小鼠胚胎干细胞衍生神经元 (ESN) 是一种可扩展的神经元模型，适用于中高通量平台，可有效用于筛选范例。通常用于通过关键神经元表型的表达进行筛选的其他转化细胞系的局限性以及通过ESN形成形态成熟和功能敏感的突触的分离神经培养物的局限性。 A� 攻击导致突触连接丧失。用 A� 治疗后，这些神经元表现出突触后蛋白 95 (PDS-95) 的显着丧失。我们建议建立并小型化检测突触/PSD-95 的方法。由于之前的研究已经确定了与 AD 表型相关的几种 PI 修饰酶，因此我们预计这种全系统方法将能够识别出突触维持的 PI 代谢途径的候选成分。改善 A 触发的突触损失的已知和新靶标以及验证高内涵筛选平台这些研究将支持未来的研究。 PI 在 Aβ 触发缺陷的细胞通路中的作用，并为使用 ESN 进行更广泛的筛查范例铺平道路。