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.

We established the pluripotent interleukin-1β (IL-1β) 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.这些蛋白质都激活了小胶质细胞和IL-1β的逐渐升高，从而驱动了AD的标志骨料的长期增强：A AD：Aβ斑块的形成，以及通过IL-1β诱导的特定激酶的合成和激活，在神经纤维中的tau蛋白质蛋白质蛋白质蛋白质和胶合膜中的多磷酸化，tau的过度磷酸化。有趣的是，在父母双方继承阿尔茨海默氏症基因的AD患者中，这两个事件均大大增强（基因型APOE4,4）。尽管这些IL-1β和APOE基因型驱动事件有利于负面结果，但我们在药物开发计划中的进展表明它们可以接受治疗。具体而言，显示GFAP结合化学剂可抑制蛋白质聚集。此外，我们发现了APOE£4基因的新功能，这是其蛋白质产物ApoE4暴露的有毒功能。我们已经证明，APOE4与转录因子EB（TFEB）竞争与透明DNA基序的结合，因此，阻碍了三种对溶酶体自噬至关重要的蛋白的转录。我们证明了AD 4,4的脑组织中的情况，但不是AD 3,3患者。现在，重要的是，我们已经确定了与APOE4蛋白结合的铅化合物，从而消除了其与透明DNA的相互作用并恢复了三个自噬基因的表达，该基因的表达编码为P62，LC3B和LAMP2蛋白的生产。现在，我们准备进一步阐明IL-1β在细胞病理生理学中的作用，建立其对激酶和激酶靶标的作用，以表现出驱动主要的聚集物成核或传播事件的修饰。我们的晚期分子和组织学方法将应用于确认从蛋白质蛋白质方法（尤其是涉及GFAP的蛋白质）相互作用的蛋白质蛋白相互作用的预测。此外，这些相互作用现在可以在大脑区域和疾病状态之间进行评估，以测试其与已知的AD参数的一致性。最后，我们将阐明针对APOE4的新型小分子药物的作用机理，以抑制其与其他APOE4靶向DNA序列的所有病理相互作用。成功完成我们提出的工作有望挫败APOE4在阿尔茨海默氏神经病发生中扮演的已知戏剧作用的预防策略。 相关性：成功完成这项提出的工作将消除阿尔茨海默基因遗传的许多病理学方面（APOEE4）在美国四分之一的人中，即8000万个遗传，即继承该基因的一个或两个副本。因此，通过特定药物候选者抑制APOE4蛋白的作用，我们将恢复有效清除大骨料所需的溶酶体自噬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，这将受到抑制。