Investigating modifier transcripts in mouse models of motor neuron disease

该项目将集中在各种疾病，总结下的术语“运动神经元疾病”，其主要病理目标是运动神经元。尽管运动神经元是这些疾病的主要目标，但并非所有运动神经元都同样容易受到病理学的影响。一些运动神经元在疾病过程中很早就受到影响和损失，而其他运动神经元受到的影响要小得多，并且保持相对完整，直到疾病的晚期。这可以在患者和动物模型中显示，包括这些疾病的小鼠模型。这些知识为本项目提供了有价值的基础，为研究调节运动神经元脆弱性的因素提供了可能性。本项目的目标是确定可以改变运动神经元脆弱性的转录本，并利用这些新获得的知识为神经保护药物的开发做出贡献。第一个目的是确定差异脆弱的运动神经元之间的潜在共性。为了实现这一目标，生物信息学将用于比较不同脆弱运动神经元的独立筛选。将分析大型数据集，进行途径分析，并基于Kline et al.（2017）中使用的类似方法，使用功能聚类生成高优先级转录物列表。此外，将分析来自中间SMA样小鼠模型的一系列肌肉的NMJ病理学，以研究它们与更严重的疾病小鼠模型相比的差异脆弱性，以及这与鉴定的感兴趣转录物的关系。第二个目的是通过敲低或过表达人类干细胞衍生的运动神经元（iPSC）中的修饰转录物来研究潜在的保护性或退行性质量和作用机制。第三个目的是研究修饰转录物在运动神经元疾病小鼠模型中的任何保护作用。将使用药理学方法和病毒基因治疗的组合来操纵在上述目的中鉴定的转录物和途径。将对神经肌肉系统进行病理学分析，包括脊髓切片和染色，以进行运动神经元计数，本项目的两位主管在实验室进行的独立研究已经观察到不同的脆弱运动神经元，并鉴定了在脆弱和抵抗运动神经元之间显示出不同表达水平的转录本。神经元（Hedlund等人，2010; Murray等人，2015）。因此，他们预测这些转录本可能是疾病的有效调节剂。本项目将进一步研究其潜在的神经保护或神经退行性质量和作用机制。

This project will focus on a variety of disorders that are summarised under the term "motor neuron disease" and whose primary pathological target are motor neurons. Even though motor neurons are the primary target of these diseases, not all motor neurons are equally vulnerable to pathology. Some motor neurons are affected and lost very early in the course of the disease, while others are much less affected and stay comparatively intact until late stages of the disease. This can be shown in both patients and in animal models, including mouse models of these diseases. This knowledge creates a valuable basis for this project, opening the possibility for the investigation of factors that regulate motor neuron vulnerability.The goal of this project is to identify transcripts that can alter motor neuron vulnerability and use this newly gained knowledge to contribute to the development of neuroprotective drugs. The first aim is to identify potential commonalities between differentially vulnerable motor neurons. In order to achieve this, bioinformatics is going to be used to compare independent screens on differentially vulnerable motor neurons. Large data sets will be analysed, pathway analysis will be performed and functional clustering will be used to generate a list of high priority transcripts, based on similar methods used in Kline et al. (2017). Furthermore, a range of muscles from an intermediate SMA-like mouse model will be analysed for NMJ pathology to investigate their differential vulnerability, in comparison to a more severe mouse models of the disease and how this relates to identified transcripts of interest. The second aim is to investigate potential protective or degenerative qualities and mechanisms of action through knock-down or overexpression of modifier transcripts in human stem cell-derived motor neurons (iPSCs). The third aim is to investigate any protective effects of modifier transcripts in a mouse model of motor neuron disease. Transcripts and pathways identified in the aims described above will be manipulated using a combination of pharmacological approaches and viral gene therapy. Pathological analysis of the neuromuscular system will be performed, which will include sectioning and staining of spinal cords to perform motor neuron counts, and dissecting and staining muscles to perform analysis of neuromuscular junctions.Independent studies in the laboratories of both supervisors of this project already observed differentially vulnerable motor neurons and identified transcripts which display an altered expression level between vulnerable and resistant motor neurons (Hedlund et al., 2010; Murray et al., 2015). They therefore predict that these transcripts could be effective modifiers of disease. This project will further investigate their potential neuroprotective or neurodegenerative qualities and mechanism of action.