LAGENBIO, Faculty of Veterinary-IIS, IA2-CITA, CIBERNED, University of Zaragoza, Zaragoza, Spain.
ALS Unit, Neurology Department, CIBERER U-723, Health Research Institute, Madrid, Spain.
Front Immunol. 2019 Apr 12;10:801. doi: 10.3389/fimmu.2019.00801. eCollection 2019.
There is growing evidence of the role of inflammation in Amyotrophic Lateral Sclerosis (ALS) during the last decade. Although the origin of ALS remains unknown, multiple potential inflammatory biomarkers have been described in ALS patients and murine models of this disease to explain the progressive motor neuron loss and muscle atrophy. However, the results remain controversial. To shed light on this issue, we aimed to identify novel biomarkers of inflammation that can influence disease progression and survival in serial blood samples from transgenic SOD1G93A mice, a model of ALS. A cytokine array assay was performed to analyze protein expression of 97 cytokines in plasma samples from wildtype controls and transgenic SOD1G93A mice at asymptomatic stage. Subsequently, serial plasma samples were obtained from SOD1G93A mice at early symptomatic, symptomatic and terminal stages to monitor cytokine levels during disease progression through immunoassays. Comparisons of means of quantifiable cytokines between short-and long-lived mice were analyzed by unrelated -test or Mann-Whitney U-test. Relationships between cytokines levels and survival time were assessed using Pearson's correlation analysis and Kaplan-Meier analysis. A total of 16 cytokines (6Ckine, ALK-1, CD30 L, eotaxin-1, galectin-1, GITR, IL-2, IL-6, IL-10, IL-13, IL-17B R, MIP-1α, MIP-3β, RANKL, TROY, and VEGF-D) were found dysregulated in transgenic SOD1G93A mice at asymptomatic stage compared with age-matched controls. Immunoassays of serial samples revealed positive expression of ALK-1, GITR and IL-17B R at P60 and P90 in mice with shorter survival. In addition, eotaxin-1 and galectin-1 levels were significantly increased at terminal stage in SOD1G93A mice that showed shorter survival time. Finally, levels of eotaxin-1, galectin-1, IL-2, IL-6, MIP-1α, and TROY at P90 or endpoint negatively correlated with the longevity of transgenic mice. We demonstrated in the SOD1G93A model of ALS that increased levels of several cytokines were associated with a shorter lifespan. However, their role as prognostic biomarkers is unclear as their expression was very variable depending on both the disease stage and the subject. Nevertheless, cytokines may be potential therapeutic targets.
在过去十年中，越来越多的证据表明炎症在肌萎缩侧索硬化症（ALS）中起作用。尽管 ALS 的病因仍不清楚，但在 ALS 患者和该疾病的小鼠模型中已描述了多种潜在的炎症生物标志物，以解释进行性运动神经元丧失和肌肉萎缩。然而，结果仍存在争议。为了阐明这一问题，我们旨在鉴定可影响转 SOD1G93A 小鼠（ALS 模型）连续血液样本中疾病进展和生存的新型炎症标志物。我们使用细胞因子阵列分析法分析了无症状期野生型对照和转 SOD1G93A 小鼠的血浆样本中 97 种细胞因子的蛋白表达。随后，通过免疫测定法从 SOD1G93A 小鼠的早期症状期、症状期和终末期获得连续的血浆样本，以监测疾病进展过程中的细胞因子水平。通过非相关 t 检验或曼-惠特尼 U 检验分析短寿命和长寿命小鼠之间可定量细胞因子的均值比较。使用 Pearson 相关分析和 Kaplan-Meier 分析评估细胞因子水平与生存时间之间的关系。与年龄匹配的对照组相比，在无症状期的转 SOD1G93A 小鼠中发现 16 种细胞因子（6Ckine、ALK-1、CD30L、嗜酸性粒细胞趋化因子-1、半乳糖凝集素-1、GITR、IL-2、IL-6、IL-10、IL-13、IL-17B R、MIP-1α、MIP-3β、RANKL、TROY 和 VEGF-D）表达失调。对连续样本的免疫测定显示，在生存期较短的 P60 和 P90 时，ALK-1、GITR 和 IL-17B R 呈阳性表达。此外，在生存期较短的 SOD1G93A 小鼠的终末期，嗜酸性粒细胞趋化因子-1 和半乳糖凝集素-1 水平显著升高。最后，在 P90 或终点时，嗜酸性粒细胞趋化因子-1、半乳糖凝集素-1、IL-2、IL-6、MIP-1α 和 TROY 的水平与转 SOD1G93A 小鼠的寿命呈负相关。我们在 SOD1G93A 型 ALS 模型中证明，几种细胞因子水平的升高与寿命缩短有关。然而，由于其表达因疾病阶段和个体而异，因此它们作为预后生物标志物的作用尚不清楚。然而，细胞因子可能是潜在的治疗靶点。
