Time-resolved single-cell analysis of neural cell differentiation in motor neuron disease

肌萎缩性侧索硬化症（ALS）是一种进行性运动神经元疾病，由上下运动神经元的选择性变性引起，最终导致瘫痪。为什么运动神经元如此脆弱，疾病相关的早期变化是如何出现的，以及其他支持细胞是如何导致疾病的，这些仍然是难以捉摸的。在这个项目中，我们将调查和描述疾病的早期阶段，同时关注特定的细胞类型和遗传背景。更具体地说，我们将利用来自两个家族性ALS队列、散发性ALS病例和年龄匹配对照的诱导多能干细胞（iPSCs）。这些多能干细胞将分化为运动神经元和星形胶质细胞，然后在细胞分化的关键阶段使用单细胞RNA测序和基于蛋白质的测定。通过这种方式，我们将能够阐明最早的疾病相关变化发生的时间，并确定与不同队列相关的关键分子变化。总之，这将使用临床相关的人体模型系统提供疾病早期阶段的时间分辨分析。此外，我们将更好地了解不同的细胞类型在哪个阶段变得脆弱，与不同的疾病相关的遗传学。然后，这些信息将用于在疾病的早期阶段测试假定的治疗干预措施。

Amyotrophic Lateral Sclerosis (ALS) is a progressive motor neuron disease that is caused by the selective degeneration of the upper and lower motor neurons that eventually leads to paralysis. It is still elusive why motor neurons are so vulnerable, how early disease associated changes appear, and precisely how other supporting cells contribute to the disease.In this project we will investigate and characterise the earliest stages of the disease while focusing on specific cell types and genetic backgrounds. More specifically, we will be utilising induced pluripotent stem cells (iPSCs) from two familial ALS cohorts, sporadic cases of ALS and age-matched controls. These iPSCs will then be differentiated into motor neurons and astrocytes, with single cell RNA sequencing and protein-based assays then employed at key stages of the cell differentiations. In this way, we will be able to elucidate when the earliest disease associated changes occur and identify the key molecular changes relevant to the different cohorts. Together this will provide a time-resolved analysis of the earliest stages of disease using a clinically relevant human model system. Moreover, we will better understand at which stage diverse cell types become vulnerable in relation to diverse disease relevant genetics. Together this information will then be used to test putative therapeutic interventions at the early stages of the disease.