Arakawa Masayuki, Arakawa Reiko, Tatsumi Shinichi, Aoki Ryoko, Saito Kayoko, Nomoto Akio
Institute of Microbial Chemistry (BIKAKEN), Tokyo, 3-14-23 Kamiosaki, Shinagawa-ku, Tokyo 141-0021, Japan.
Institute of Medical Genetics, Tokyo Women's Medical University, 10-22 Kawadacho, Shinjyuku-ku, Tokyo 162-0054, Japan.
Biochem Biophys Res Commun. 2014 Oct 24;453(3):368-74. doi: 10.1016/j.bbrc.2014.09.087. Epub 2014 Sep 27.
Spinal muscular atrophy (SMA) is caused by mutations within the survival motor neuron 1 (SMN1) gene. These mutations result in the reduction of survival motor neuron (SMN) protein expression and SMN complex in spinal motor neurons and other tissues. SMN protein has been used as a therapeutic biomarker in recent SMA clinical studies using enzyme-linked immunosorbent assay (ELISA). Here, we investigated whether imaging flow cytometry can be a viable source of quantitative information on the SMN protein. Using a FlowSight imaging flow cytometer (Merck-Millipore, Germany), we demonstrated that imaging flow cytometry could successfully identify different expression patterns and subcellular localization of SMN protein in healthy human fibroblasts and SMA patient-derived fibroblasts. In addition, we could also evaluate the therapeutic effects of SMN protein expression by valproic acid treatment of SMA patient-derived cells in vitro. Therefore, we suggest that imaging flow cytometry technology has the potential for identifying SMN protein expression level and pattern as an evaluation tool of clinical studies.
脊髓性肌萎缩症（SMA）由生存运动神经元1（SMN1）基因突变引起。这些突变导致脊髓运动神经元和其他组织中生存运动神经元（SMN）蛋白表达及SMN复合物减少。在近期使用酶联免疫吸附测定（ELISA）的SMA临床研究中，SMN蛋白已被用作治疗生物标志物。在此，我们研究了成像流式细胞术是否能成为获取SMN蛋白定量信息的可行来源。使用FlowSight成像流式细胞仪（德国默克密理博公司），我们证明成像流式细胞术能够成功识别健康人成纤维细胞和SMA患者来源的成纤维细胞中SMN蛋白的不同表达模式和亚细胞定位。此外，我们还可以通过体外丙戊酸处理SMA患者来源的细胞来评估SMN蛋白表达的治疗效果。因此，我们认为成像流式细胞术技术有潜力作为临床研究的评估工具来识别SMN蛋白表达水平和模式。
