Evaluating autophagy modulation as a therapeutic strategy for Alzheimer's Disease using human neuronal culture models

使用人类神经元培养模型评估自噬调节作为阿尔茨海默病的治疗策略

• 批准号: 10750709
• 负责人: Amanda Snead
• 金额: $ 5.27万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-16 至 2027-08-15
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750709
• 关键词: Abeta synthesis; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Amyloid beta-42; Amyloid beta-Protein; Amyotrophic Lateral Sclerosis; Autophagocytosis; Autophagosome; Axon; Axonal Transport; Clinical; Comparative Study; Competitive Binding; Complementary therapies; Data; Dimensions; Disease; Distal; Drug Design; Drug Targeting; FDA approved; Frontotemporal Dementia; Functional disorder; Hela Cells; Human; Image; Immunoprecipitation; Impaired cognition; Impairment; Investigation; Lysosomes; Measures; Membrane Proteins; Modeling; Morphology; Movement; Neurites; Neurodegenerative Disorders; Neurons; Organelles; Parkinson Disease; Pathogenesis; Pathologic; Pathology; Pathway interactions; Pharmaceutical Preparations; Positioning Attribute; Process; Proteins; Proteomics; Route; Sirolimus; Swelling; Testing; Therapeutic; Vesicle; abeta accumulation; cell motility; cognitive function; effective therapy; efficacy evaluation; experimental study; extracellular; familial Alzheimer disease; high throughput screening; improved; induced pluripotent stem cell; insight; misfolded protein; neuronal cell body; novel; novel therapeutics; protein aggregation; protein degradation; retrograde transport; synergism; targeted treatment; translational potential

ABSTRACT To date there is no effective treatment for Alzheimer’s disease that decreases cognitive decline. Although the available drugs are effective at reducing amyloid beta, a multi-target drug approach is more likely to succeed in impacting cognitive function. One potentially complementary treatment approach is modulation of the autophagic lysosomal pathway (ALP), which is responsible for protein turnover and has been shown to regulate degradation of misfolded proteins in the CNS. Autophagy activation normally helps clear protein aggregates and damaged organelles via sequestration into autophagosomes, these vesicles eventually fuse with lysosomes, which then degrade the autophagic cargo. This pathway is more complicated in long-lived, polarized neurons, where autophagy largely initiates in the distal axon and thus autophagic lysosomal intermediates must be removed from the axon via retrograde transport to the soma where proteolytically active lysosomes reside. Disruptions in the transport or maturation of ALP intermediates has long been implicated in AD pathogenesis, as they buildup in dystrophic neurites around Aβ aggregates and likely contribute to Aβ production in AD. Preliminary data in our AD model of iPSC-derived human i3Neurons shows that ALP intermediates that accumulate in axonal swellings are cleared upon treatment with a novel compound identified in a high throughput screen as an autophagy upregulator in Hela cells. Importantly, our data shows that this compound also reduces both intracellular and extracellular Aβ42 and increases neuronal autophagy (increased LC3II/LC3I) in our AD model i3Neurons. Exciting new data from the Aldrich lab has identified lysosomal membrane protein 1 (LAMP1) as a direct target of the compound. Given these preliminary studies, we hypothesize that the novel compound, through direct interaction with LAMP1 and potential stabilization of LAMP1 interactions with retrograde machinery, increases retrograde lysosomal transport and autophagosome maturation, and thus clears axonal ALP vesicles and ultimately lowers Aβ levels. We will test this hypothesis by determining the mechanism by which the novel compound alters ALP transport and ALP composition and function, as well as identify new potential targets in neurons (by an unbiased proteomics approach). Through these studies, we will also determine if the FDA-approved drug Rapamycin can alter axonal ALP buildup or Aβ42 levels in comparison to the novel compound, thus shedding new insight into the translational potential of both drugs. Lastly, we will determine if the novel compound can reduce Aβ42 in a familial AD model and how it alters ALP in these neurons. Given the strong evidence that this compound can modulate axonal ALP transport, Aβ42 and neuronal autophagy, the results from the proposed experiments could be relevant to therapeutic approaches in other neurodegenerative diseases that have ALP dysfunction as a contributing pathological feature, such as Parkinson’s disease, amyotrophic lateral sclerosis, and frontotemporal dementia.

抽象的 迄今为止，还没有有效的治疗阿尔茨海默病的方法可以减少认知能力的下降。 现有药物可有效减少β淀粉样蛋白，多靶点药物方法更有可能成功 影响认知功能的一种潜在的补充治疗方法是调节自噬。 溶酶体途径 (ALP)，负责蛋白质周转并已被证明可以调节降解 中枢神经系统中错误折叠的蛋白质的自噬激活通常有助于清除蛋白质聚集体和受损的蛋白质。 通过封存在自噬体中的细胞器，这些囊泡最终与溶酶体融合，然后 降解自噬货物的途径在长寿的极化神经元中更为复杂，其中 自噬主要在远端轴突开始，因此必须去除自噬溶酶体中间产物 从轴突通过逆行运输到达具有蛋白水解活性的溶酶体所在的胞体。 随着 ALP 中间体的积累，ALP 中间体的运输或成熟长期以来一直与 AD 发病机制有关 在 Aβ 聚集周围的营养不良性神经突中，可能有助于 AD 中 Aβ 的产生。 iPSC 衍生的人类 i3Neurons 的 AD 模型表明 ALP 中间体在轴突肿胀中积累 经过高通量筛选中鉴定为自噬的新型化合物处理后被清除 重要的是，我们的数据表明，这种化合物还可以减少细胞内和细胞内的细胞因子。 在我们的 AD 模型 i3Neurons 中，细胞外 Aβ42 并增加神经元自噬（增加 LC3II/LC3I）。 Aldrich 实验室的令人兴奋的新数据已确定溶酶体膜蛋白 1 (LAMP1) 为直接靶标 鉴于这些初步研究，我们通过直接研究得出了该新化合物。 与 LAMP1 的相互作用以及 LAMP1 与逆行机制相互作用的潜在稳定性， 增加逆行溶酶体转运和自噬体成熟，从而清除轴突 ALP 我们将通过确定其机制来检验这一假设。 该新型化合物改变了 ALP 转运以及 ALP 组成和功能，并发现了新的潜力 通过这些研究，我们还将确定神经元中的目标（通过无偏见的蛋白质组学方法）。 FDA 批准的药物雷帕霉素与新药相比可以改变轴突 ALP 积累或 Aβ42 水平 化合物，从而为这两种药物的转化潜力提供新的见解。最后，我们将确定是否可以。 这种新型化合物可以减少家族性 AD 模型中的 Aβ42 以及它如何改变这些神经元中的 ALP。 强有力的证据表明该化合物可以调节轴突 ALP 运输、Aβ42 和神经元自噬 拟议实验的结果可能与其他神经退行性疾病的治疗方法相关 以 ALP 功能障碍为病理特征的疾病，例如帕金森病， 肌萎缩侧索硬化症和额颞叶痴呆。