Autophagy control of human neuroinflammatory signalling pathways

神经炎症是衰老和/或病变大脑的一个显著特征。它可以在人类神经退行性疾病的早期检测到，并且可以在动物疾病模型中阻断以阻止神经元衰退。神经炎症是由星形胶质细胞和小胶质细胞引发和控制的。这些神经胶质细胞最初对神经元具有保护作用，但在慢性神经炎症期间逐渐变得“反应性”和毒性。我们想要了解大脑特定区域神经炎症的分子控制，以便在衰老和疾病期间控制它以保护大脑。利用分化为脑区域特异性星形胶质细胞和神经元的人诱导多能干细胞（hiPSCs），我们可以在体外复制神经炎症的各个方面。在本项目中，我们将通过将hiPSC星形胶质细胞和神经元与小胶质细胞单层和类器官共培养来推进这项工作。我们将重点关注神经炎症反应中神经胶质应激和自噬（一个重要的细胞质质量控制过程）的作用。这将涉及分泌组蛋白组学、免疫印迹、细胞因子ELISA、qRT-PCR和细胞成像，结合涉及CRISPRi和CRISPRa的自噬操作，以确定在神经炎症信号传导过程中起作用的关键分子及其对神经胶质生物学和神经元健康的下游影响。

Neuroinflammation is a prominent feature of the ageing and/or diseased brain. It can be detected early in human neurodegenerative disease, and can be blocked to halt neuronal decline in animal models of disease. Neuroinflammation is triggered and controlled by astrocytes and microglia. These glial cells are initially protective to neurons, but become progressively 'reactive' and toxic during chronic neuroinflammation. We want to understand the molecular control of neuroinflammation in specific regions of the brain, so that it might be controlled to protect the brain during ageing and disease. Using human induced pluripotent stem cells (hiPSCs) that are differentiated to become brain region-specific astrocytes and neurons, we can replicate aspects of neuroinflammation in vitro.In this project, we will advance this work by combining hiPSC astrocytes and neurons with microglia in monolayer and organoid co-cultures. We will focus on the roles of glial stress and autophagy (a crucial cytoplasmic quality control process) during the neuroinflammatory response. This will involve secretome proteomics, immunoblotting, cytokine ELISA, qRT-PCR, and cell imaging, combined with autophagy manipulation involving CRISPRi and CRISPRa, to establish the key molecular players acting during neuroinflammatory signalling and their downstream impact on glial biology and neuronal health.