Neuron Glia Biology in Health and Disease Group, Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003, Lisbon, Portugal.
Department of Biochemistry and Human Biology, Faculty of Pharmacy, Universidade de Lisboa, Lisbon, Portugal.
Mol Neurobiol. 2018 May;55(5):4207-4224. doi: 10.1007/s12035-017-0631-2. Epub 2017 Jun 13.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of unknown cause. Absence of specific targets and biomarkers compromise the development of new therapeutic strategies and of innovative tools to stratify patients and assess their responses to treatment. Here, we investigate changes in neuroprotective-neuroinflammatory actions in the spinal cord of SOD1 mice, at presymptomatic and symptomatic stages to identify stage-specific biomarkers and potential targets. Results showed that in the presymptomatic stage, there are alterations in both astrocytes and microglia, which comprise decreased expression of GFAP and S100B and upregulation of GLT-1, as well as reduced expression of CD11b, M2-phenotype markers, and a set of inflammatory mediators. Reduced levels of Connexin-43, Pannexin-1, CCL21, and CX3CL1 further indicate the existence of a compromised intercellular communication. In contrast, in the symptomatic stage, increased markers of inflammation became evident, such as NF-κB/Nlrp3-inflammasome, Iba1, pro-inflammatory cytokines, and M1-polarizion markers, together with a decreased expression of M2-phenotypic markers. We also observed upregulation of the CX3CL1-CX3CR1 axis, Connexin-43, Pannexin-1, and of microRNAs (miR)-124, miR-125b, miR-146a and miR-21. Reduced motor neuron number and presence of reactive astrocytes with decreased GFAP, GLT-1, and GLAST further characterized this inflammatory stage. Interestingly, upregulation of miR-155 and downregulation of MFG-E8 appear as consistent biomarkers of both presymptomatic and symptomatic stages. We hypothesize that downregulated cellular interplay at the early stages may represent neuroprotective mechanisms against inflammation, SOD1 aggregation, and ALS onset. The present study identified a set of inflamma-miRNAs, NLRP3-inflammasome, HMGB1, CX3CL1-CX3CR1, Connexin-43, and Pannexin-1 as emerging candidates and promising pharmacological targets that may represent potential neuroprotective strategies in ALS therapy.
肌萎缩侧索硬化症（ALS）是一种病因不明的致命神经退行性疾病。缺乏特定的靶点和生物标志物，会影响新的治疗策略的开发，也影响创新工具的开发，这些工具用于对患者进行分层，并评估他们对治疗的反应。在这里，我们研究了 SOD1 小鼠脊髓中的神经保护-神经炎症作用在无症状和有症状阶段的变化，以确定特定于阶段的生物标志物和潜在靶点。结果表明，在无症状阶段，星形胶质细胞和小胶质细胞都发生了改变，包括 GFAP 和 S100B 的表达下调，GLT-1 的上调，以及 CD11b、M2 表型标志物和一组炎症介质的表达下调。Connexin-43、Pannexin-1、CCL21 和 CX3CL1 的水平降低进一步表明细胞间通讯受损。相比之下，在有症状阶段，炎症标志物明显增加，如 NF-κB/Nlrp3 炎性小体、Iba1、促炎细胞因子和 M1 极化标志物，同时 M2 表型标志物的表达下调。我们还观察到 CX3CL1-CX3CR1 轴、Connexin-43、Pannexin-1 和 microRNAs（miR）-124、miR-125b、miR-146a 和 miR-21 的上调。运动神经元数量减少，反应性星形胶质细胞减少，GFAP、GLT-1 和 GLAST 减少，进一步说明了这一炎症阶段的特征。有趣的是，miR-155 的上调和 MFG-E8 的下调似乎是无症状和有症状阶段的一致生物标志物。我们假设，早期细胞相互作用的下调可能代表了针对炎症、SOD1 聚集和 ALS 发作的神经保护机制。本研究确定了一组炎症 microRNAs、NLRP3 炎性小体、HMGB1、CX3CL1-CX3CR1、Connexin-43 和 Pannexin-1，它们是新兴的候选药物靶点和有前途的药理学靶点，可能代表 ALS 治疗中的潜在神经保护策略。
