Connor Sarah M, Rashid Mamunur, Ryan Katie J, Patel Kruti, Boyd Justin D, Smith Jennifer, Elyaman Wassim, Bennett David A, Bradshaw Elizabeth M
Columbia University Irving Medical Center, New York, NY, United States.
Ann Romney Center for Neurologic Diseases, Harvard Medical School, Brigham and Women's Hospital, Boston, MA, United States.
Front Cell Neurosci. 2022 May 23;16:894601. doi: 10.3389/fncel.2022.894601. eCollection 2022.
Microglia, the resident immune cells of the central nervous system (CNS), are responsible for maintaining homeostasis in the brain by clearing debris and are suggested to be inefficient in Alzheimer's Disease (AD), a progressive neurodegenerative disorder for which there is no disease-modifying drug. Besides pathological approaches, unbiased evidence from genome-wide association studies (GWAS) and gene network analysis implicate genes expressed in microglia that reduce phagocytic ability as susceptibility genes for AD. Thus, a central feature toward AD therapy is to increase the microglial phagocytic activities while maintaining synaptic integrity. Here, we developed a robust unbiased high content screening assay to identify potential therapeutics which can reduce the amyloid-beta (Aβ1-42) load by increasing microglial uptake ability. Our screen identified the small-molecule GW5074, an inhibitor of c-RAF, a serine/threonine kinase, which significantly increased the Aβ1-42 clearance activities in human monocyte-derived microglia-like (MDMi) cells, a microglia culture model that recapitulates many genetic and phenotypic aspects of human microglia. Notably, GW5074 was previously reported to be neuroprotective for cerebellar granule cells and cortical neurons. We found that GW5074 significantly increased the expression of key AD-associated microglial molecules known to modulate phagocytosis: TYROBP, SIRPβ1, and TREM2. Our results demonstrated that GW5074 is a potential therapeutic for AD, by targeting microglia.
小胶质细胞是中枢神经系统(CNS)中的常驻免疫细胞,负责通过清除碎片来维持大脑的内环境稳定。在阿尔茨海默病(AD)这种尚无疾病修饰药物的进行性神经退行性疾病中,小胶质细胞的功能被认为是低效的。除了病理学方法外,全基因组关联研究(GWAS)和基因网络分析的无偏证据表明,小胶质细胞中表达的降低吞噬能力的基因是AD的易感基因。因此,AD治疗的一个核心特点是在维持突触完整性的同时增强小胶质细胞的吞噬活性。在这里,我们开发了一种强大的无偏倚高内涵筛选试验,以鉴定能够通过提高小胶质细胞摄取能力来降低β淀粉样蛋白(Aβ1-42)负荷的潜在治疗药物。我们的筛选鉴定出了小分子GW5074,它是一种丝氨酸/苏氨酸激酶c-RAF的抑制剂,可显著增强人单核细胞衍生的小胶质细胞样(MDMi)细胞中的Aβ1-42清除活性,MDMi细胞是一种模拟人类小胶质细胞许多遗传和表型特征的小胶质细胞培养模型。值得注意的是,此前有报道称GW5074对小脑颗粒细胞和皮质神经元具有神经保护作用。我们发现GW5074显著增加了已知可调节吞噬作用的关键AD相关小胶质细胞分子的表达:TYROBP、SIRPβ1和TREM2。我们的结果表明,GW5074通过靶向小胶质细胞是一种潜在的AD治疗药物。