Dulbecco Telethon Institute, Milano, Italy.
PLoS One. 2011;6(10):e25545. doi: 10.1371/journal.pone.0025545. Epub 2011 Oct 5.
Amyotrophic lateral sclerosis (ALS) is a fatal progressive motor neuron disease, for which there are still no diagnostic/prognostic test and therapy. Specific molecular biomarkers are urgently needed to facilitate clinical studies and speed up the development of effective treatments.
METHODOLOGY/PRINCIPAL FINDINGS: We used a two-dimensional difference in gel electrophoresis approach to identify in easily accessible clinical samples, peripheral blood mononuclear cells (PBMC), a panel of protein biomarkers that are closely associated with ALS. Validations and a longitudinal study were performed by immunoassays on a selected number of proteins. The same proteins were also measured in PBMC and spinal cord of a G93A SOD1 transgenic rat model. We identified combinations of protein biomarkers that can distinguish, with high discriminatory power, ALS patients from healthy controls (98%), and from patients with neurological disorders that may resemble ALS (91%), between two levels of disease severity (90%), and a number of translational biomarkers, that link responses between human and animal model. We demonstrated that TDP-43, cyclophilin A and ERp57 associate with disease progression in a longitudinal study. Moreover, the protein profile changes detected in peripheral blood mononuclear cells of ALS patients are suggestive of possible intracellular pathogenic mechanisms such as endoplasmic reticulum stress, nitrative stress, disturbances in redox regulation and RNA processing.
CONCLUSIONS/SIGNIFICANCE: Our results indicate that PBMC multiprotein biomarkers could contribute to determine amyotrophic lateral sclerosis diagnosis, differential diagnosis, disease severity and progression, and may help to elucidate pathogenic mechanisms.
肌萎缩侧索硬化症（ALS）是一种致命的进行性运动神经元疾病，目前仍没有诊断/预后测试和治疗方法。迫切需要特定的分子生物标志物来促进临床研究并加速有效治疗方法的开发。
方法/主要发现：我们使用二维差异凝胶电泳方法，在容易获得的临床样本外周血单核细胞（PBMC）中鉴定出与 ALS 密切相关的一组蛋白质生物标志物。通过对选定数量的蛋白质进行免疫测定进行验证和纵向研究。同样的蛋白质也在 G93A SOD1 转基因大鼠模型的 PBMC 和脊髓中进行了测量。我们确定了蛋白质生物标志物的组合，可以以高区分能力区分 ALS 患者与健康对照（98％），以及与可能类似于 ALS 的神经障碍患者（91％），区分两种疾病严重程度（90％），以及许多转化生物标志物，这些标志物将人与动物模型之间的反应联系起来。我们证明 TDP-43、亲环蛋白 A 和 ERp57 在纵向研究中与疾病进展相关。此外，在 ALS 患者的外周血单核细胞中检测到的蛋白质谱变化表明可能存在细胞内致病机制，如内质网应激、硝化应激、氧化还原调节和 RNA 处理的紊乱。
结论/意义：我们的研究结果表明，PBMC 多种蛋白质生物标志物可能有助于确定肌萎缩侧索硬化症的诊断、鉴别诊断、疾病严重程度和进展，并可能有助于阐明致病机制。
