Metabolomic Profiling to Identify Molecular Biomarkers of Cellular Response to Methotrexate In Vitro
Metabolomic Profiling to Identify Molecular Biomarkers of Cellular Response to Methotrexate In Vitro
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DOI：
10.1111/cts.12694
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发表时间：
2019-10-25
影响因子：
3.9
通讯作者：
Becker, Mara L.
中科院分区：
文献类型：
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作者：
Funk, Ryan S.;Singh, Rakesh K.;Becker, Mara L.
在自身免疫性关节炎的治疗中，甲氨蝶呤(MTX)疗效的差异是早期有效控制疾病的重要障碍。我们推测，代谢组学技术的应用将有助于更好地了解MTX的药理活性的生化基础，并可以促进MTX反应的新的分子生物标志物的识别和评估。在这项工作中，红系细胞暴露于生理浓度为1,000 nM的MTX，并使用三个代谢组平台进行分析，以获得广泛的细胞代谢物。MTX的药理活性，定义为抑制细胞生长，根据对724种已鉴定代谢物的分析，与细胞代谢谱的改变有关。通过判别分析，MTX治疗与酮异戊酸、果糖、半乳糖和2-脱氧胞苷增加有关，而2-脱氧尿苷、磷脂酰肌醇32：0、玫瑰花酸和肌苷一磷酸相应减少。将叶酸代谢分析的数据与化学计量学和代谢网络分析相结合，表明MTX治疗与叶酸代谢和核苷酸生物合成失调有关，这与其已知的作用机制是一致的。然而，MTX治疗也与多种代谢物的改变有关，包括氨基酸、碳水化合物和脂肪代谢的中间体。总体而言，这些发现支持在暴露于生理浓度的MTX后出现强劲的代谢反应。它们还确定了与MTX药理活性相关的各种代谢中间体，因此，在未来MTX治疗自身免疫性关节炎的临床前和临床研究中，它们是潜在的分子生物标记物。
Variation in methotrexate (MTX) efficacy represents a significant barrier to early and effective disease control in the treatment of autoimmune arthritis. We hypothesize that the utilization of metabolomic techniques will allow for an improved understanding of the biochemical basis for the pharmacological activity of MTX, and can promote the identification and evaluation of novel molecular biomarkers of MTX response. In this work, erythroblastoid cells were exposed to MTX at the physiologic concentration of 1,000 nM and analyzed using three metabolomic platforms to give a broad spectrum of cellular metabolites. MTX pharmacological activity, defined as cellular growth inhibition, was associated with an altered cellular metabolomic profile based on the analysis of 724 identified metabolites. By discriminant analysis, MTX treatment was associated with increases in ketoisovaleric acid, fructose, galactose, and 2-deoxycytidine, and corresponding reductions in 2-deoxyuridine, phosphatidylinositol 32:0, orotic acid, and inosine monophosphate. Inclusion of data from analysis of folate metabolism in combination with chemometric and metabolic network analysis demonstrated that MTX treatment is associated with dysregulated folate metabolism and nucleotide biosynthesis, which is in line with its known mechanism of action. However, MTX treatment was also associated with alterations in a diversity of metabolites, including intermediates of amino acid, carbohydrate, and lipid metabolism. Collectively, these findings support a robust metabolic response following exposure to physiologic concentrations of MTX. They also identify various metabolic intermediates that are associated with the pharmacological activity of MTX, and are, therefore, potential molecular biomarker candidates in future preclinical and clinical studies of MTX efficacy in autoimmune arthritis.