Modelling peripheral axonal pathology of ALS in vitro - Assembly of optogenetic neuromuscular circuits from pluripotent stem cells

体外模拟 ALS 的外周轴突病理学 - 多能干细胞光遗传学神经肌肉回路的组装

• 批准号: MR/N025865/1
• 负责人: Ivo Lieberam
• 金额: $ 61.38万
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FN025865%2F1
• 关键词: Modelling; peripheral; axonal; pathology; ALS

Amyotrophic Lateral Sclerosis (ALS) is a medical condition which affects motor neurons, the type of nerve cell which connects the brain to muscle. ALS leads to paralysis, so that affected individuals can no longer walk, speak or breathe. The disease is both quite common and currently incurable, and therefore many scientists study its causes and possible therapies. As many manipulations required to investigate ALS cannot be carried out in patients, experimental animals, mostly mice and rats, are used in most studies. These animals are often genetically modified such that they develop a disease which is similar to the one seen in humans. This approach uses a large number of animals and is expensive and time-consuming, and therefore not suitable to systematically study hereditary traits which may cause ALS. We propose to develop an alternative way of investigating ALS, which will be based on complex, life-like artificial tissues grown directly from cultured human stem cells. The artificial tissues will resemble nerve tissue (motor neurons and accessory cells called 'glia') and muscle. They will be cultured in a compartmentalized biochip designed and manufactured for this project. In this device, the two tissues will be segregated - as they are in humans and animals-, and the motor neurons will connect to muscle through nerve fibres and control muscle contractions. The cells in the device will contain biological optical probes that allow us to feed instructions into the cells on the biochip and read information out though fluorescent light signals. We will test the validity of the new experimental system by first modelling the degeneration of nerves and the loss of communication between nerves and muscle. Then, we will use this 'ALS in a dish' model to investigate why different forms of ALS result in different patterns of degeneration, how genetic traits protect some people from developing ALS, and how such protection may be mimicked with chemical compounds. We think that this tissue culture model could allow scientist to discover ALS drugs faster, cheaper and without the use of animal experimentation.

肌萎缩侧索硬化症(ALS)是一种会影响运动神经元的疾病，运动神经元是连接大脑和肌肉的神经细胞类型。肌萎缩侧索硬化症会导致瘫痪，因此受影响的人不能再走路、说话或呼吸。这种疾病很常见，目前还无法治愈，因此许多科学家研究了它的原因和可能的治疗方法。由于研究肌萎缩侧索硬化症所需的许多操作不能在患者身上进行，因此大多数研究都使用实验动物，主要是小鼠和大鼠。这些动物通常是经过基因改造的，因此它们会患上一种与人类相似的疾病。这种方法使用大量的动物，昂贵和耗时，因此不适合系统地研究可能导致ALS的遗传性状。我们建议开发一种研究肌萎缩侧索硬化症的替代方法，这种方法将基于直接从培养的人类干细胞生长出来的复杂的、类似生命的人工组织。人造组织将类似于神经组织(运动神经元和被称为胶质细胞的附属细胞)和肌肉。他们将在为该项目设计和制造的隔间生物芯片中进行培养。在这个装置中，这两个组织将被分离--就像在人类和动物身上一样--运动神经元将通过神经纤维连接到肌肉，并控制肌肉收缩。该装置中的细胞将包含生物光学探头，允许我们将指令输入生物芯片上的细胞，并通过荧光信号读出信息。我们将通过首先模拟神经退变和神经与肌肉之间通讯的丧失来测试新实验系统的有效性。然后，我们将使用这个“盘子中的肌萎缩侧索硬化症”模型来研究为什么不同形式的肌萎缩侧索硬化症会导致不同的退化模式，遗传特征如何保护一些人免受肌萎缩侧索硬化症的发展，以及这种保护可能如何被化合物模仿。我们认为，这种组织培养模式可以让科学家更快、更便宜地发现ALS药物，而且不需要使用动物实验。

项目成果

Biobased Elastomer Nanofibers Guide Light-Controlled Human-iPSC-Derived Skeletal Myofibers.

• DOI: 10.1002/adma.202110441
• 发表时间: 2022-05
• 期刊: Advanced materials (Deerfield Beach, Fla.)

3D Compartmentalised Human Pluripotent Stem Cell-derived Neuromuscular Co-cultures.

3D分室的人多能干细胞衍生的神经肌肉共培养。

• DOI: 10.21769/bioprotoc.4624
• 发表时间: 2023-03-05
• 期刊: Bio-protocol
• 影响因子: 0.8

Optogenetic modeling of human neuromuscular circuits in Duchenne muscular dystrophy with CRISPR and pharmacological corrections.

• DOI: 10.1126/sciadv.abi8787
• 发表时间: 2021-09-10
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Paredes-Redondo A;Harley P;Maniati E;Ryan D;Louzada S;Meng J;Kowala A;Fu B;Yang F;Liu P;Marino S;Pourquié O;Muntoni F;Wang J;Lieberam I;Lin YY
• 通讯作者: Lin YY

Enrichment of human embryonic stem cell-derived V3 interneurons using an Nkx2-2 gene-specific reporter.

• DOI: 10.1038/s41598-023-29165-z
• 发表时间: 2023-02-03
• 期刊: Scientific reports
• 影响因子: 4.6