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: 10673064
• 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/10673064
• 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; Data; Defect; Dementia; Dementia with Lewy Bodies; Development; Development Plans; Diabetes Mellitus; Elderly; Endosomes; Energy Metabolism; Energy consumption; 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; Knowledge; Lead; Lipids; Measures; Mentorship; Metabolic; Metabolic Pathway; Metabolic dysfunction; Metabolic stress; Metabolism; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Parkinson Disease; Pathology; Pathway interactions; Persons; Predisposition; Property; Proteins; Radiolabeled; Recycling; Research; Research Personnel; Respiration; Risk Factors; Societies; Stress; Substrate Interaction; Supplementation; Synapses; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Intervention; Training; Ubiquitin; Variant; alpha synuclein; apolipoprotein E-3; apolipoprotein E-4; career development; causal variant; cell growth regulation; disorder risk; effective therapy; experience; functional decline; gene interaction; 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 cell body; 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.

项目概要 神经退行性痴呆，包括阿尔茨海默病，会造成毁灭性的认知能力下降，因此 代谢应激是导致痴呆症的原因之一：葡萄糖无法治愈。 代谢低下是阿尔茨海默病 (AD) 和帕金森病 (PD) 的早期特征，而糖尿病是 然而，代谢压力如何与遗传相结合。 导致神经元死亡的神经退行性疾病危险因素尚不清楚，并且没有已知的 为了解决这些知识差距，我们已经确定了增强易感神经元代谢弹性的方法。 维持细胞能量水平的遗传目标，使用结合切割的独特筛选范例 用于 ATP 的边缘基因编码传感器，ATP 是细胞中的主要能量携带分子，具有全基因组 通过这种方法，我们已经确定了基于 CRISPR 的基因通路。 当细胞处于代谢压力下时，对 ATP 水平的显着影响尤其是我们的初步数据。 表明敲除与内吞再循环相关的 AD/PD 疾病风险基因会导致神经元死亡 特别是当葡萄糖缺乏时，其中一个基因是 SORL1，这是一种可能导致 AD 的风险基因。 SORL1 已知与 APOE 相互作用，而 APOE4 变异是 AD 的最大遗传风险因素。 APOE 和 SORL1 之间的相互作用或内吞回收途径尚不清楚。 Ken Nakamura 博士和 Robert Mahley 博士的指导，以及 Martin Kampmann 博士和 托马斯·格雷伯 (Thomas Graeber)，并得到了格拉德斯通研究所充满活力的研究社区和核心的支持 和加州大学旧金山分校，我将检验以下假设：内吞循环中的 AD 风险突变会导致能量衰竭，并且 增加神经元对脂质缺乏和 APOE 表达的有害影响的敏感性 我将通过以下目标研究这一假设：1) 确定 APOE4 基因型的影响。 破坏内吞循环对能量消耗和呼吸的影响，2) 确定受损的影响 葡萄糖代谢对内体蛋白运输的影响，3) 确定 APOE 表达和基因型 导致内吞回收缺陷神经元的代谢脆弱性和内体蛋白运输。 拟议的研究还将确定是否通过基因操作或解决问题来维持 ATP 水平 代谢缺陷具有治疗神经元脆弱性和功能衰退的治疗潜力。 拟议的研究和职业发展计划将建立在我之前的培训基础上并增强我的发展轨迹 通过培养研究能量代谢和蛋白质稳态的技能，成为一名独立研究者 在单个神经元中，掌握分析技术来研究代谢功能障碍和代谢反应 强调系统层面，获得实验设计、沟通和指导方面的经验，以及 为痴呆和神经退行性疾病病理生理学的生物学理解奠定基础。