Inducing neuroprotective responses in astrocytes using human stem cell models

使用人类干细胞模型诱导星形胶质细胞的神经保护反应

• 批准号: 2250134
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2250134
• 关键词: Inducing; neuroprotective; responses; astrocytes; using

Amyotrophic lateral sclerosis (ALS) is a debilitating neurodegenerative disorder characterised by progressive loss of motor neurons in the spinal cord, brain stem and motor cortex. While research has predominantly focused on neuronal involvement in ALS, recent studies have explored non-cell autonomous pathomechanisms and highlighted the complex interplay between motor neurons and non-neuronal cells in the CNS, such as astrocytes. Under normal conditions, astrocytes play myriad supportive roles in maintaining brain homeostasis. In neurological diseases, such as ALS and ischaemic stroke, astrocytes undergo a phenotypic switch to become reactive, thus acquiring potential to exert both pro- and anti-inflammatory properties. These states have been coined A1 (neurotoxic) and A2 (neuroprotective) respectively, however astrocyte reactivity is more likely a heterogenous spectrum rather than two distinct states. While the A1 state has been characterised in vitro, an A2 state is yet to be functionally validated. Although hypoxia is thought to facilitate neurodegeneration, known markers of protective reactivity are expressed by astrocytes under moderate hypoxic conditions. Against this background, my PhD will first establish a human induced pluripotent stem cell model of (functionally validated) neuroprotective astrocytes using a range of established assays and some that I will develop myself. Where assays require developing, all preliminary work will be performed in HEK293s for experimental tractability. I will next address the molecular mechanisms by which human astrocytes mount neuroprotective responses by comparing the transcriptome and proteome of protective astrocytes with already established models of basal and toxic/deleterious astrocytes. I will finally systematically examine how neuroprotective mechanisms are disrupted in ALS. Crucially, the directed differentiation paradigm together with functional assays, co-culture assays and other essential 'know-how' is already established and routinely performed in my host lab, which mitigates risk and allows me to focus on my specific biological questions.

肌萎缩侧索硬化症（ALS）是一种以脊髓、脑干和运动皮质中的运动神经元的进行性丧失为特征的使人衰弱的神经退行性疾病。虽然研究主要集中在ALS中的神经元参与，但最近的研究探索了非细胞自主病理机制，并强调了CNS中运动神经元和非神经元细胞（如星形胶质细胞）之间的复杂相互作用。在正常情况下，星形胶质细胞在维持脑内环境稳定方面发挥着无数的支持作用。在神经系统疾病中，如ALS和缺血性中风，星形胶质细胞经历表型转换成为反应性的，从而获得发挥促炎和抗炎特性的潜力。这些状态分别被称为A1（神经毒性）和A2（神经保护），然而星形胶质细胞的反应性更可能是一种异质性谱，而不是两种不同的状态。虽然A1状态已在体外表征，但A2状态尚未在功能上得到验证。虽然缺氧被认为是促进神经退行性变，已知的保护性反应标记物在中度缺氧条件下由星形胶质细胞表达。在此背景下，我的博士学位将首先使用一系列已建立的测定方法和一些我自己开发的方法建立一个（功能验证的）神经保护性星形胶质细胞的人类诱导多能干细胞模型。如果需要开发检测试剂盒，则将在HEK 293中进行所有初步工作，以确保实验的易处理性。接下来，我将通过比较保护性星形胶质细胞的转录组和蛋白质组与已经建立的基础和有毒/有害星形胶质细胞模型来解决人类星形胶质细胞产生神经保护反应的分子机制。最后，我将系统地研究ALS中神经保护机制是如何被破坏的。至关重要的是，定向分化模式以及功能测定，共培养测定和其他必要的“专有技术”已经在我的主机实验室建立并定期进行，这降低了风险，并使我能够专注于我的特定生物学问题。