Imaging Flow Cytometry Analysis to Identify Differences of Survival Motor Neuron Protein Expression in Patients With Spinal Muscular Atrophy
Imaging Flow Cytometry Analysis to Identify Differences of Survival Motor Neuron Protein Expression in Patients With Spinal Muscular Atrophy
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DOI：
10.1016/j.pediatrneurol.2016.05.009
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发表时间：
2016-08-01
影响因子：
3.8
通讯作者：
Saito, Kayoko
中科院分区：
文献类型：
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作者：
Arakawa, Reiko;Arakawa, Masayuki;Saito, Kayoko
背景技术背景：脊髓性肌萎缩症是由运动神经元中运动神经元存活蛋白表达不足引起的神经退行性疾病。疾病改善疗法的主要目标是增加运动神经元存活表达。运动神经元存活蛋白表达的变化可以通过患者的外周血细胞进行监测，因此我们测试了成像流式细胞术的灵敏度和实用性。方法：在分别用两个和三个SMN 2基因拷贝的人健康对照受试者和两个1型脊髓性肌萎缩症患者的外周血淋巴细胞永生化后，我们使用成像流式细胞术分析来确定存活运动神经元表达的显著差异。明亮的细节强度分析被用来研究运动神经元存活蛋白的细胞定位的差异。研究结果：相对于来自健康对照受试者的那些，来自脊髓性肌萎缩症患者的细胞中的存活运动神经元表达显著降低。此外，根据SMN 2拷贝数，运动神经元存活表达与脊髓性肌萎缩症的临床严重程度相关。相对于来自健康对照受试者的那些，来自脊髓性肌萎缩症患者的细胞中存活运动神经元蛋白的细胞积累也显著降低。结论：用于外周血分析的成像流式细胞术的益处包括其分析异质细胞群体的能力;可视化细胞形态;以及评估靶蛋白的积累、定位和表达。在未来的研究中，应实施成像流细胞学分析，以优化其作为脊髓性肌萎缩症临床试验工具的应用。
BACKGROUND: Spinal muscular atrophy is a neurodegenerative disorder caused by the deficient expression of survival motor neuron protein in motor neurons. A major goal of disease-modifying therapy is to increase survival motor neuron expression. Changes in survival motor neuron protein expression can be monitored via peripheral blood cells in patients; therefore we tested the sensitivity and utility of imaging flow cytometry for this purpose. METHODS: After the immortalization of peripheral blood lymphocytes from a human healthy control subject and two patients with spinal muscular atrophy type 1 with two and three copies of SMN2 gene, respectively, we used imaging flow cytometry analysis to identify significant differences in survival motor neuron expression. A bright detail intensity analysis was used to investigate differences in the cellular localization of survival motor neuron protein. RESULTS: Survival motor neuron expression was significantly decreased in cells derived from patients with spinal muscular atrophy relative to those derived from a healthy control subject. Moreover, survival motor neuron expression correlated with the clinical severity of spinal muscular atrophy according to SMN2 copy number. The cellular accumulation of survival motor neuron protein was also significantly decreased in cells derived from patients with spinal muscular atrophy relative to those derived from a healthy control subject. CONCLUSIONS: The benefits of imaging flow cytometry for peripheral blood analysis include its capacities for analyzing heterogeneous cell populations; visualizing cell morphology; and evaluating the accumulation, localization, and expression of a target protein. Imaging flow 'cytorhetry analysis should be 'implemented in future studies to optimize its' application as a tool for spinal muscular atrophy clinical trials.