Defective lysosomal membrane fission mediates axonal lysosome accumulation in dystrophic neurites in Alzheimer's disease.

溶酶体膜裂变缺陷介导阿尔茨海默病营养不良神经突中轴突溶酶体的积累。

• 批准号: 9812548
• 负责人: Matthew Schrag
• 金额: $ 24.3万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9812548
• 关键词: Abeta clearance; Adult; Age-Months; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease risk; Amyloid beta-Protein; Amyloidosis; Animal Model; Area; Axon; Behavioral; Biology; Blood Vessels; Brain; Caregiver Burden; Caregivers; Cell physiology; Cells; Cerebral Amyloid Angiopathy; Cerebrum; Clinical Management; Clinical Trials; Code; Cognition; Cognitive; Cognitive aging; Communities; Complex; Confocal Microscopy; Coupled; Custom; Dementia; Deposition; Deterioration; Development; Disease; Elderly; Electron Microscopy; Environment; Enzymes; Epidemic; Failure; Functional disorder; Genes; Genetic Models; Healthcare; Hela Cells; Human; Image Analysis; Immune system; Impaired cognition; Impairment; International; Investigational Therapies; Knock-out; Lead; Learning; Lysosomes; Mediating; Membrane; Membrane Fusion; Memory; Mentorship; Molecular; Mus; Mutate; Mutation; Nerve Degeneration; Neurites; Neurobiology; Neurons; Outcome; Pathogenesis; Pathologic; Pathology; Performance; Phenotype; Phospholipase; Phospholipase D; Phospholipases A; Plasmids; Process; Proteins; Public Health; Publishing; Regulation; Reporting; Research; Resources; Role; Running; Senile Plaques; System; Techniques; Testing; Therapeutic; Tissue Model; Training; Variant; WFS1 gene; Work; abeta deposition; aging brain; animal model development; base; brain tissue; career development; cerebral amyloidosis; cohort; cost; density; effective therapy; enzyme activity; genetic risk factor; human tissue; improved; in vitro Assay; in vivo; link protein; lysosome membrane; mouse model; neurobehavioral; neuropathology; new therapeutic target; novel; programs; protein aggregate; protein function; proteostasis; quantitative imaging; selective expression; skills; stem

Alzheimer’s disease (AD) compromises the independence of aging adults and is associated with huge societal burden in healthcare and caregiver costs. An effective treatment is urgently needed to stem the tide of the ongoing and growing epidemic of dementia. Many recent clinical trials have attempted to reduce the level of β- amyloid in the brain, typically by activating the immune system to promote clearance, yet none of these trials has been successful. We propose to explore this problem in a different way – we discovered that neuronal axons around deposits of β-amyloid are swollen and filled with abnormal lysosomes which are deficient in protein-degrading enzymes. Because lysosomes are critical for protein homeostasis, we hypothesize that these abnormal lysosomes contribute to neurodegeneration and if their function could be rescued, it could improve brain function. We will study this hypothesis by focusing on a novel gene, PLD3, which was identified as contributing to AD risk. In our preliminary work, we discovered that PLD3 is robustly enriched on these abnormal lysosomes and brain PLD3 levels correlated inversely with both β-amyloid burden and the rate of cognitive deterioration in a human cohort. Brain PLD3 levels also correlated inversely with memory performance in a mouse model. We discovered that PLD3 functions as a phospholipase D in acidic environments and is necessary for lysosomal membrane fission. We propose to evaluate to what degree the lysosome dysfunction observed in mice is present in human tissue and to fully evaluate the association of PLD3 with neuronal lysosomal pathology. We will then determine whether the coding variants reported to confer AD-risk impact the function of PLD3 by transfecting plasmids containing PLD3 (which have been mutated to copy these coding variants) and observing the effect on lysosomal membrane fission and on PLD3 enzyme activity. We will determine whether defective neuronal lysosomal fission impacts cognition and AD neuropathology in vivo by crossing the 5xFAD model of genetic AD with a conditional knock-out of Fig4 using a Cre selectively expressed in cerebral neurons. Fig4 is a component of the PIKfyve complex and loss of Fig4 leads to defective lysosomal fission. Importantly, the PIKfyve complex also regulates PLD3 processing and alters PLD3 activity. We will evaluate learning and memory in this mouse and determine whether defective lysosomal fission impacts β-amyloidosis and dystrophic neurites. These studies will help us understand both how this protein functions in the setting of AD and the role of lysosome membrane fusion in cognitive aging and AD. The parallel training plan will support my training in model animal development and behavioral phenotyping, developing advanced skills in lyosomal neurobiology and refine clinical management of aging related diseases, all of which are critical steps for my career development. Collectively, the outstanding institutional environment, resources, and interdisciplinary mentorship team with broad and complementary areas of expertise will accelerate my acquisition of the necessary expertise to transition to independence.

阿尔茨海默病（AD）损害老年人的独立性并与庞大的社会有关 医疗保健和护理人员费用的负担迫切需要一种有效的治疗方法来遏制这种趋势。 痴呆症的流行持续且日益严重，最近许多临床试验都试图降低β-水平。 大脑中的淀粉样蛋白，通常通过激活免疫系统来促进清除，但这些试验都没有 我们建议以不同的方式探索这个问题——我们发现了神经元。 β-淀粉样蛋白沉积周围的轴突肿胀并充满异常溶酶体，这些溶酶体缺乏 由于溶酶体对于蛋白质稳态至关重要，因此我们对此进行了努力。 这些异常的溶酶体会导致神经退行性变，如果它们的功能能够得到挽救， 我们将通过关注一种新基因 PLD3 来研究这一假设，该基因已被鉴定出来。 在我们的初步工作中，我们发现 PLD3 在这些方面得到了极大的丰富。 异常的溶酶体和大脑 PLD3 水平与 β-淀粉样蛋白负荷和发生率呈负相关。 人类大脑 PLD3 水平的认知恶化也与记忆力呈负相关。 我们发现 PLD3 在酸性条件下起到磷脂酶 D 的作用。 我们建议评估溶酶体膜分裂的程度。 在小鼠中观察到的溶酶体功能障碍也存在于人体组织中，并充分评估与 然后我们将确定 PLD3 是否与神经元溶酶体病理有关。 通过转染含有 PLD3 的质粒（已被 突变以复制这些编码变体）并观察对溶酶体膜裂变和 PLD3 的影响 我们将确定缺陷的神经溶酶体裂变是否会影响认知和 AD。 通过将遗传性 AD 的 5xFAD 模型与 Fig4 的条件敲除相结合，进行体内神经病理学研究 Cre 在大脑神经元中选择性表达，是 PIKfyve 复合体的组成部分，而 Fig4 缺失。 导致溶酶体分裂缺陷，重要的是，PIKfyve 复合物还调节 PLD3 加工和 我们将评估这只小鼠的学习和记忆能力，并确定是否存在缺陷。 溶酶体裂变影响β-淀粉样变性和营养不良性神经突，这些研究将帮助我们了解这两者。 这种蛋白质如何在 AD 中发挥作用以及溶酶体膜融合在认知衰老中的作用 并行训练计划将支持我在模型动物发育和行为方面的训练。 表型分析、发展溶酶体神经生物学的先进技能和完善衰老的临床管理 相关疾病，所有这些都是我职业发展的关键步骤。 制度环境、资源和跨学科导师团队广泛且互补 专业领域将加速我获得必要的专业知识以过渡到独立。