Characterisation of the motor neurons obtained from induced pluripotent stem cells (iPS) in Amyotrophic lateral sclerosis (ALS).

肌萎缩侧索硬化症(ALS)是一种严重的神经退行性疾病，累及上、下运动神经元(MN)，确诊后2-3年内可导致死亡。我们之前的发现和确定ALS患者MN转录图谱的可能性以及对其功能特征的研究促使我们与同济大学的徐军等人在中国建立了合作关系。徐军教授和高正良教授在同济大学干细胞研究中心建立了非常好的干细胞研究设施。他们在干细胞研究方面拥有国际公认的跟踪记录，并成功地从人类成纤维细胞中培养出运动神经元样细胞。这项合作是为了有效地推进ALS研究的共同努力，融合了技术专业知识和大量生物样本。应用基因芯片技术分析肌萎缩侧索硬化症(ALS)患者和对照组诱导的多能干细胞(IPS)来源的运动神经元(MN)，将揭示肌萎缩侧索硬化症(ALS)的基因和通路异常。这项研究计划将阐明MN的选择性脆弱性，从而能够确定潜在的治疗靶点。本研究的目的是：1.从中国同济大学的合作者那里获得iPS来源的MN，并在英国谢菲尔德的宿主实验室建立这项技术。2.确定iPS来源的MN的表达谱3.比较散发性ALS患者和家族性ALS患者的MN表达谱与对照组的差异。4.比较患者和对照组MN的应激反应，并比较同一个体MN和成纤维细胞的应激反应。5.鉴定iPS来源的ALS细胞在培养液中释放的细胞因子和其他因子及其对对照组MN的影响。6.比较患者来源的MN和对照来源的MN的轴突运输特性。7.测试能够拮抗或诱导通过上述调查确定的相关途径的药物。

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disorder affecting upper and lower motoneurons (MN) and leading to death within 2-3 years from diagnosis. Our previous findings and the possibility to determine the transcription profile of MN derived from ALS patients as well as investigating their functional characteristics led us to set up a collaboration with Professor Jun Xu et al at Tongji University in China. Professor Jun Xu and Professor Zhengliang Gao have established very good stem cell research facility at Stem Cell Research Center at Tongji University. They have international recognised track records in stem cell research and have successfully produced motor neuron like cells from human fibroblast cells. This collaboration is a joint effort to efficiently progress in ALS research, merging technical expertise and a large number of biological samples. The application of microarray analysis to the induced pluripotent stem cells (iPS) derived motor neurones (MN) from patients and controls will uncover the genes and pathways dysregulated in Amyotrophic lateral sclerosis (ALS). This research programme will elucidate the selective vulnerability of MN allowing the identification of potential therapeutic targets. The aims of the proposed study are: 1. To produce iPS-derived MN from our collaborators at Tongji University in China and set up the technique in the host laboratory in Sheffield, UK. 2. To determine the expression profile of the iPS-derived MN 3. To compare the expression profile of MN derived from sporadic and familial ALS patients with control. 4. To determine the stress response of MN obtained from patients compared to controls as well as comparing the stress response of MN and fibroblasts from the same individual 5. To identify the cytokines and other factors released in the media by the iPS-derived ALS cells and their effect on control MN. 6. To compare the axonal transport characteristics of patient derived MN with control derived MN. 7. To test drugs able to antagonise or induce relevant pathways identified through the investigations outlined above.