Metabolic Vulnerability and Effects of APOE in Human Neurons with Impaired Endocytic Recycling

内吞循环受损的人类神经元的代谢脆弱性和 APOE 的影响

• 批准号: 10507737
• 负责人: Neal Bennett
• 金额: $ 13.1万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10507737
• 关键词: Address; Alzheimer' s Disease; Alzheimer' s disease risk; Amyloid beta-Protein; Biological; CRISPR interference; Cell Death; Cell Line; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Communication; Communities; Consumption; Data; Defect; Dementia; Dementia with Lewy Bodies; Development; Development Plans; Diabetes Mellitus; Elderly; Endosomes; Energy Metabolism; Experimental Designs; Exposure to; Failure; Foundations; Functional disorder; Genes; Genetic; Genetic Predisposition to Disease; Genotype; Glucose; Glycolysis; Goals; Human; Impaired cognition; Impairment; Individual; Induced pluripotent stem cell derived neurons; Institutes; Knowledge; Lead; Lipids; Measures; Mentorship; Metabolic; Metabolic Pathway; Metabolic dysfunction; Metabolic stress; Metabolism; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Parkinson Disease; Pathology; Pathway interactions; Persons; Predisposition; Property; Proteins; Radiolabeled; Recycling; Research; Research Personnel; Respiration; Risk Factors; Societies; Stress; Supplementation; Synapses; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Intervention; Training; Ubiquitin; Variant; alpha synuclein; base; career development; causal variant; cell growth regulation; disorder risk; effective therapy; experience; functional decline; gene therapy; genetic manipulation; genetic risk factor; glucose metabolism; improved; induced pluripotent stem cell; insight; knock-down; metabolic abnormality assessment; metabolomics; neurodegenerative dementia; neuron loss; neuronal survival; novel; preservation; prevent; programs; protein transport; proteostasis; research and development; resilience; response; risk variant; screening; sensor; skills; tau Proteins; transcriptomics; whole genome

PROJECT SUMMARY Neurodegenerative dementias, including Alzheimer’s disease, inflict devastating cognitive decline, for which there is no cure. Metabolic stress is hypothesized to contribute to the development of dementia: glucose hypometabolism is an early feature in Alzheimer’s (AD) and Parkinson’s disease (PD), and diabetes is a substantial risk factor for developing dementia. However, how metabolic stress combines with genetic neurodegenerative disease risk factors to lead to neuronal death is not well understood, and there are no known ways to boost metabolic resilience in susceptible neurons. To address these knowledge gaps, we have identified genetic targets that maintain cellular energy levels, using a unique screening paradigm that combines cutting- edge, genetically encoded sensors for ATP, the main energy-carrying molecule in cells, with whole-genome CRISPR-based gene manipulations. With this approach, we have identified gene pathways that have a prominent impact on ATP levels when cells are under metabolic stress. In particular, our preliminary data indicate that knockdown of AD/PD disease risk genes associated with endocytic recycling lead to neuronal death specifically when glucose is scarce. One of these genes is SORL1, a risk gene that is potentially causal for AD. SORL1 is known to interact with APOE, and the APOE4 variant is the largest genetic risk factor for AD. But the interaction between APOE and SORL1 or the endocytic recycling pathway is not well understood. Under the mentorship of Dr. Ken Nakamura and Dr. Robert Mahley, in collaboration with Drs. Martin Kampmann and Thomas Graeber, and with the support of the vibrant research community and cores at the Gladstone Institutes and UCSF, I will test the hypothesis that AD risk mutations in endocytic recycling create an energy failure and increases the susceptibility of neurons to lipid deficits and the deleterious effects of APOE expression and APOE4 genotype. I will investigate this hypothesis through the following Aims: 1) Determine the impact of disrupting endocytic recycling on energy consumption and respiration, 2) Determine the effect of impaired glucose metabolism on endosomal protein trafficking, 3) Determine how APOE expression and genotype contribute to metabolic vulnerability and endosomal protein trafficking in endocytic recycling deficient neurons. The proposed studies will also determine if maintaining ATP levels via genetic manipulations or by addressing metabolic deficits have therapeutic potential for treating neuron vulnerability and functional decline. The proposed research and career development plan will build on my previous training and enhance my trajectory toward becoming an independent investigator by developing skills to study energy metabolism and proteostasis in individual neurons, mastering analytical techniques to study metabolic dysfunction and response to metabolic stress on a systems-level, gaining experience with experimental design, communication, and mentorship, and building a foundation of biological understanding of dementia and neurodegenerative disease pathophysiology.

项目摘要 神经退行性痴呆症，包括阿尔茨海默氏病，导致毁灭性的认知下降，为此 没有治愈方法。 缺乏代谢是阿尔茨海默氏症（AD）和帕金森氏病（PD）的早期特征，糖尿病是一种 但是，痴呆症的大量风险因素。 神经退行性疾病危险因素导致神经元死亡尚不清楚，在这里 在感知神经元中的Toost代谢弹性。 遗传靶标的thargets tarular能量水平 边缘，遗传编码的ATP传感器，ATP是细胞中的主要能量携带分子，具有全基因组 基于CRISPR的基因操作。 当细胞尤其是代谢应力时，对ATP水平的显着影响。 表明与内吞和内吞作用相关的AD/PD疾病风险基因的敲低导致神经元死亡 具体来说，当葡萄糖稀缺时，这些基因之一是SORL1，这是一种潜在的AD因果关系。 已知SORL1与APOE相互作用，APOE4变体是地址的最大遗传因素。 APOE和SORL1或内吞回收途径之间的相互作用尚不清楚。 Ken Nakamura博士和Robert Mahley博士的指导与Martin Kampmann博士合作 托马斯·格雷伯（Thomas Graeber），在格拉德斯通学院充满活力的研究委员会和核心的支持下 和UCSF，我将检验以下假设：内吞娱乐中的AD风险突变会导致能量失败和 增加神经元对脂质缺陷和ApoE表达有害的敏感性 APOE4基因型。 破坏内吞回收对能耗和呼吸的回收，2）确定受损的影响 内体蛋白运输上的葡萄糖代谢，3）确定APOE表达和基因型如何 有助于内吞回收型神经元中的代谢脆弱性和内体蛋白运输。 具有支撑性研究还确定是通过遗传操作维持ATP水平还是通过解决 代谢缺陷具有治疗神经元脆弱性和功能下降的治疗潜力 支撑的研究和职业发展计划将以我以前的培训为基础，并增强我的轨迹 通过发展技能tudy能量代谢和蛋白质的技能来成为恒定的独立研究者 在单个神经元中，掌握分析技术来研究代谢功能障碍和对代谢的反应 对系统级别的压力，在实验设计，沟通和指导方面获得经验 建立对痴呆症和神经退行性疾病病理生理学的生物学理解的基础。