Neuroinflammation, Protein Aggregates, ApoE4 Drug Targeting, and Autophagy Rescue

神经炎症、蛋白质聚集体、ApoE4 药物靶向和自噬拯救

• 批准号: 10768318
• 负责人: Sue Tilton Griffin
• 金额: $ 47.03万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10768318
• 关键词: Acute-Phase Proteins; Alzheimer' s Disease; Alzheimer' s neuropathogenesis; Amyloid beta-Protein; Apolipoprotein E; Astrocytes; Autophagocytosis; Binding; Brain; Brain Diseases; Brain region; Chemical Agents; Chronic; DNA; DNA Sequence; Development; Drug Targeting; Event; Exhibits; Feedback; Functional disorder; Genes; Genetic Transcription; Genotype; Glial Fibrillary Acidic Protein; Histologic; Individual; Inherited; Interleukin-1 beta; Lead; Microglia; Modification; Molecular; Motion; Neurofibrillary Tangles; Neurons; Outcome; Parents; Pathogenesis; Patients; Pharmaceutical Preparations; Phosphotransferases; Play; Prevention strategy; Production; Protein Inhibition; Proteins; Proteomics; Reporting; Repression; Role; Stress; Testing; Work; apolipoprotein E-4; brain tissue; chemical binding; cytokine; drug candidate; gain of function; hyperphosphorylated tau; neuroinflammation; novel; protein aggregation; protein protein interaction; small molecule; transcription factor

We established the pluripotent interleukin-1β (IL-1 β) cytokine as a significant player in the pathogenesis of Alzheimer’s disease as it sets in motion a self-amplifying positive-feedback cycle in which neuronal stress induces synthesis of the neuron's acute phase protein β APP and release of its fragments sAPPa and Aβ. Both these proteins activate microglia and a progressive elevation of IL-1 β, which drives chronically enhanced formation of the hallmark aggregates of AD: A β plaques and, via IL-1 β -induced synthesis and activation of specific kinases, hyperphosphorylation of tau in neurofibrillary tangles in neurons and glial fibrillary acidic protein (GFAP) in astrocytes. Interestingly, both events are dramatically enhanced in AD patients who inherit the Alzheimer gene from both parents (genotype ApoE4,4). Although these IL-1 β and ApoE genotype driven events favor negative outcomes, our progress in drug-development initiatives shows that they are amenable to treatment. Specifically, GFAP-binding chemical agents were shown to inhibit protein aggregation. Further, we discovered a novel function of the APOE£4 gene, which is a toxic gain-of- function exhibited by its protein product, ApoE4. We have demonstrated that ApoE4 competes with Transcriptional Factor EB (TFEB) for binding to the CLEAR DNA motifs, thus, hindering the transcription of three proteins crucial for lysosomal autophagy. We show this to be the case in brain tissues from AD 4,4, but not AD 3,3 patients. Now, importantly, we have identified a lead compound that binds to ApoE4 protein, obviating its interactions with CLEAR DNA and restoring the expression of three autophagy genes, that encode for production of p62, LC3B, and LAMP2 proteins. Now, we are prepared to further elucidate the role of IL-1 β in cellular pathophysiology, establishing its effects on kinases and kinase targets that manifest the modifications which drive predominant aggregate nucleation or propagation events. Our advanced molecular and histological approaches will be applied to confirm predictions of protein-protein interactions derived from proteomics approaches, particularly those involving GFAP. Moreover, these interactions can now be evaluated across brain regions and disease states to test their concordance with known AD parameters. Finally, we will elucidate the mechanisms of action of our identified novel small-molecule drugs targeted to ApoE4 in inhibiting all its pathognomonic interactions with other ApoE4 targeted DNA sequences. Successful completion of our proposed work promises a preventive strategy for foiling the known dramatic role that ApoE4 plays in Alzheimer neuropathogenesis. RELEVANCE: Successful completion of this proposed work will obviate the many pathognomonic aspects of inheritance of the Alzheimer gene (APOEE4) in the 1 in 4 individuals in the US, i.e., 80 million, who inherit one or both copies of this gene. Through the action of our specific drug candidates to inhibit the ApoE4 protein we will, therefore, restore the lysosomal autophagy necessary for efficient clearance of large aggregates, which, as we reported, is repressed in the brains of those who inherit one or both APOE4 genes.

我们确定多能白细胞介素 1β (IL-1 β) 细胞因子是阿尔茨海默病发病机制中的重要参与者，因为它启动自我放大的正反馈循环，其中神经元应激诱导神经元期急性蛋白 β 的合成APP 及其片段 sAPPa 和 Aβ 的释放这两种蛋白都会激活小胶质细胞和 IL-1 β 的逐渐升高，从而驱动 AD 标志性聚集体的长期增强形成：A。 β 斑块，以及通过 IL-1 β 诱导的特定激酶的合成和激活、神经元神经原纤维缠结中 tau 蛋白的过度磷酸化以及星形胶质细胞中隐含的神经胶质原纤维酸性蛋白 (GFAP)，这两种事件在遗传性 AD 患者中均显着增强。来自父母双方的阿尔茨海默病基因（基因型 ApoE4,4） 尽管这些 IL-1 β 和 ApoE 基因型驱动的事件有利于负面结果，但我们在药物开发计划方面的进展表明具体而言，GFAP 结合化学剂被证明可以抑制蛋白质聚集。此外，我们还发现了 APOE£4 基因的一种新功能，即其蛋白质产物 ApoE4 所表现出的毒性功能获得。我们已经证明 ApoE4 与转录因子 EB (TFEB) 竞争与 CLEAR DNA 基序的结合，从而阻碍对溶酶体自噬至关重要的三种蛋白质的转录。 AD 4,4 患者的脑组织中存在这种情况，但 AD 3,3 患者的脑组织中并非如此。现在，重要的是，我们已经鉴定出一种与 ApoE4 蛋白结合的先导化合物，从而消除了其与 CLEAR DNA 的相互作用，并恢复了三个自噬基因的表达。 ，编码 p62、LC3B 和 LAMP2 蛋白的产生现在，我们准备进一步阐明 IL-1 β 在细胞病理生理学中的作用，确定其对激酶和激酶的影响。我们先进的分子和组织学方法将用于确认来自蛋白质组学方法的蛋白质-蛋白质相互作用的预测，特别是那些涉及 GFAP 的方法。最后，我们将阐明我们确定的针对 ApoE4 的新型小分子药物在抑制其与其他药物的所有特征性相互作用方面的作用机制。 ApoE4 靶向 DNA 序列的成功完成我们提出的工作有望提供一种预防策略，以阻止 ApoE4 在阿尔茨海默病神经发病机制中发挥的已知的戏剧性作用。 相关性：这项拟议工作的成功完成将消除美国四分之一（即 8000 万人）遗传阿尔茨海默基因 (APOEE4) 的许多特征。因此，我们的特定候选药物抑制 ApoE4 蛋白的作用将恢复有效清除大聚集物所必需的溶酶体自噬，正如我们报道的那样，在继承一个或两个 APOE4 基因的人的大脑中受到抑制。