源于肌萎缩侧索硬化症iPSC运动神经元的分化与干细胞康复电生理研究

Amyotrophic lateral sclerosis (ALS) is a lethal disease involving the loss of motor neurons but the mechanism is unclear. Using the technology of induced pluripotent stem cell (iPSC), we can get cell line of the same genetic background with the ALS patient, which can be then differentiated to motor neurons that repeat the pathological state of the patient and can be repeatedly used for the disease mechanism study. We have proved that iPSCs from patients with degenerative nerve disease like Spinal Muscular Atrophy can be successfully differentiated into neuron in vitro. In this study, the similar methods will be used to reprogram patient skin fibroblasts into iPSCs and then direct differentiate into neurons. Through molecular cell biology study and patch clamp recording, we will try to identify and describe the differences of the electrophysiological function between neurons from patient and normal human iPSCs, including the characterization of action potential and the currents of the voltage gated channels, especially the function of motor neurons. Taking together, our research is to first establish the familial ALS disease cell model using patient-derived iPSCs, which can be further used to study the disease mechanism and examine the drug effect. This study will help us to have a better understanding of the fundamental mechanism underlying the clinical phenotype and clinical ALS therapy.

SOD1位点突变引起运动神经元的丢失，导致肌萎缩侧索硬化症的发生，但机制不明。我们前期研究证明了源于病人的iPSC可分化为成熟的神经元。进一步实验发现这些神经元有电活性异常和离子通道电流变化。那么，源于ALS病人iPSC分化的神经元的电生理发生了哪些变化？ 由此，我们提出新的思路，利用iPSC技术可获得与ALS病人遗传背景完全相同的细胞系，其定向分化的神经元就能完整再现ALS的病理状态。本项目在前期实验的基础上，将利用该项新技术将ALS病人皮肤成纤维细胞"变身"为iPSC并定向分化为神经元。采用干细胞实验技术及膜片钳电生理记录方法，研究源于正常人和ALS病人的iPSCS定向分化得到的神经元电生理功能，包括动作电位特征和电压门控离子通道活性，特别是运动神经元功能变化。旨在建立病人来源的ALS疾病细胞模型，获得电生理数据。本项目有望阐明ALS发病机制，还将有助于康复治疗ALS药物的筛选。

SOD1位点突变引起运动神经元的丢失，导致肌萎缩侧索硬化症的发生，但机制不明。我们前期研究证明了源于病人的iPSC可分化为成熟的神经元。进一步实验发现这些神经元有电活性异常和离子通道电流变化。那么，源于ALS病人iPSC分化的神经元的电生理发生了哪些变化？ 由此，我们提出新的思路，利用iPSC技术可获得与ALS病人遗传背景完全相同的细胞系，其定向分化的神经元就能完整再现ALS的病理状态。本项目在前期实验的基础上，将利用该项新技术将ALS病人皮肤成纤维细胞"变身"为iPSC并定向分化为神经元。采用干细胞实验技术及膜片钳电生理记录方法，研究源于正常人和ALS病人的iPSCS定向分化得到的神经元电生理功能，包括动作电位特征和电压门控离子通道活性，特别是运动神经元功能变化。旨在建立病人来源的ALS疾病细胞模型，获得电生理数据。本项目有望阐明ALS发病机制，还将有助于康复治疗ALS药物的筛选。