Research Service, Bruce W. Carter Veterans Affairs Medical Center, 1201 NW 16th Street, Miami, FL, 33125, USA.
Department of Neurology, University of Miami Miller School of Medicine, 1120 NW 14th St, Miami, FL, 33136, USA.
J Neurovirol. 2017 Dec;23(6):808-824. doi: 10.1007/s13365-017-0571-7. Epub 2017 Sep 14.
Fingolimod (FTY720), a structural analogue of sphingosine, targets sphingosine-1-phosphate receptor signaling and is currently an immunomodulatory therapy for multiple sclerosis. Fingolimod accesses the central nervous system (CNS) where its active metabolite, fingolimod phosphate (FTY720-P), has pleotropic neuroprotective effects in an inflammatory microenvironment. To investigate potential neuronal-specific mechanisms of fingolimod neuroprotection, we cultured the human neuronal progenitor cell line, hNP1, in differentiation medium supplemented with HIV- or Mock-infected supernatants, with or without FTY720-P. Gene expression was investigated using microarray and functional genomics. FTY720-P treatment increased differentially expressed (DE) neuronal genes by 33% in HIV-exposed and 40% in Mock-exposed cultures. FTY720-P treatment broadened the functional profile of DE genes in HIV-exposed versus Mock-exposed neurons, including not only immune responses but also transcriptional regulation and cell differentiation, among others. FTY720-P treatment downregulated the gene for follistatin, the antagonist of activin signaling, in all culture conditions. FTY720-P treatment differentially affected both glycolysis-related and immune response genes in Mock- or HIV-exposed cultures, significantly upregulating 11 glycolysis-related genes in HIV-exposed neurons. FTY720-P treatment also differentially upregulated genes related to innate immune responses and antigen presentation in Mock-exposed and more so in HIV-exposed neurons. However, in HIV-exposed neurons, FTY720-P depressed the magnitude of differential expression in almost half the genes, suggesting an anti-inflammatory potential. Moreover, in HIV-exposed neurons, FTY720-P reduced expression of the amyloid precursor protein (APP) gene, resulting in reduced expression of the APP protein. This study provides new evidence that fingolimod alters neuronal gene expression in inflammatory, viral-infected microenvironments, with the potential for neuroprotective effects.
芬戈莫德（FTY720）是一种鞘氨醇的结构类似物，靶向鞘氨醇-1-磷酸受体信号通路，目前是多发性硬化症的免疫调节治疗药物。芬戈莫德可进入中枢神经系统（CNS），其活性代谢物芬戈莫德磷酸（FTY720-P）在炎症微环境中具有多种神经保护作用。为了研究芬戈莫德神经保护的潜在神经元特异性机制，我们在含有 HIV 或模拟感染上清液的分化培养基中培养人神经祖细胞系 hNP1，同时添加或不添加 FTY720-P。使用微阵列和功能基因组学研究基因表达。FTY720-P 处理使 HIV 暴露和模拟暴露培养物中差异表达（DE）神经元基因分别增加 33%和 40%。FTY720-P 处理拓宽了 HIV 暴露与模拟暴露神经元中 DE 基因的功能谱，不仅包括免疫反应，还包括转录调控和细胞分化等。FTY720-P 处理下调了所有培养条件下激活素信号拮抗剂滤泡抑制素的基因表达。FTY720-P 处理在模拟或 HIV 暴露培养物中均差异影响糖酵解相关和免疫反应基因，显著上调 HIV 暴露神经元中 11 个糖酵解相关基因。FTY720-P 处理还差异上调了模拟暴露和更显著的 HIV 暴露神经元中与先天免疫反应和抗原呈递相关的基因。然而，在 HIV 暴露神经元中，FTY720-P 使近一半基因的差异表达幅度降低，表明具有抗炎潜力。此外，在 HIV 暴露神经元中，FTY720-P 降低了淀粉样前体蛋白（APP）基因的表达，导致 APP 蛋白表达降低。本研究提供了新的证据，表明芬戈莫德改变了炎症、病毒感染微环境中神经元的基因表达，具有神经保护作用。
