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，还没有有效的阿尔茨海默氏病治疗方法可以降低认知能力下降。虽然 可用的药物可有效减少淀粉样蛋白β，多目标药物方法更有可能成功 影响认知功能。一种潜在的完整治疗方法是调节自噬 溶酶体途径（ALP），该途径负责蛋白质周转率，并已证明调节降解 中枢神经系统中错误折叠的蛋白质。自噬激活通常有助于清除蛋白质聚集体并损坏 细胞器通过隔离为自噬体，这些蔬菜最终与溶酶体融合，然后将其融合 降解自噬货物。该途径在长寿命，两极化的神经元中更为复杂，其中 自噬大部分在远端轴突中启动，因此必须去除自噬的溶酶体中间体 从轴突通过逆行转运到蛋白水解活性溶酶体的SOMA。中断 ALP中间体的运输或成熟长期以来一直在AD发病机理中实施 在Aβ聚集体周围的营养不良神经运动中，可能有助于AD中的Aβ产生。我们的初步数据 IPSC衍生的人I3Neurons的AD模型表明，ALP中间位于轴突肿胀中 用在高吞吐量屏幕中鉴定出的新型化合物的治疗后，被清除为自噬 HeLa细胞中的上调。重要的是，我们的数据表明，该化合物还降低了细胞内和 细胞外Aβ42并在我们的AD型i3neurons中增加神经元自噬（增加LC3II/LC3I）。 来自Aldrich Lab的令人兴奋的新数据已将溶酶体膜蛋白1（LAMP1）确定为直接目标 该化合物。鉴于这些初步研究，我们假设新颖的化合物通过直接 与LAMP1的相互作用以及LAMP1相互作用与逆行机制的潜在稳定， 增加逆行溶酶体转运和自噬体成熟，从而清除轴突ALP 囊泡并最终降低了Aβ水平。我们将通过确定该假设来确定该假设的机制 新颖的复合改变了ALP的传输和ALP组成和功能，并确定了新的潜力 神经元中的靶标（通过公正的蛋白质组学方法）。通过这些研究，我们还将确定是否 与小说相比 化合物，从而对两种药物的翻译潜力进行了新的见解。最后，我们将确定是否 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。鉴于 有力的证据表明，该化合物可以调节轴突ALP转运Aβ42和神经元自噬，即 提出的实验的结果可能与其他神经退行性的治疗方法有关 具有ALP功能障碍的疾病是造成病理性特征的疾病，例如帕金森氏病， 肌萎缩性侧索硬化和额颞痴呆。