Understanding the mechanisms of ferroptosis in direct neuronal reprogramming – towards improving neuronal replacement in vivo

了解直接神经元重编程中铁死亡的机制 â 改善体内神经元替代

• 批准号: 461629173
• 负责人: Professorin Dr. Magdalena Götz
• 金额: --
• 依托单位: Institut für Physiologische Genomik
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/461629173?language=en
• 关键词: Understanding; mechanisms; ferroptosis; direct; neuronal

Neurons lost upon brain injury or in neurodegenerative disease cannot be replaced in most brain regions of mammals. We have pioneered the novel approach of direct neuronal reprogramming from local glial cells and identified ferroptosis as a major hurdle in this process. Here we aim to further dissect the pathways involved in eliciting ferroptosis when astrocytes of murine and human origin are converted into neurons in vitro. We will further explore a possible link to the unfolded protein response and analyze the role of neuron-specific antioxidant proteins identified in our recent mitochondrial proteome of astrocytes and neurons. Based on in vitro most promising results, we aim to inhibit these pathways specifically when converting reactive astrocytes after stab wound injury in the murine cerebral cortex in vivo, e.g. by overexpressing FSP1, Gpx4, deleting Acsl4 or using CRISPRa to activate the expression of neuron-specific mitochondrial proteins. These genetic manipulations will be compared to the effects of pharmacological treatments, e.g. by using inhibitors like of 3rd generation liproxstatin or Rosiglitazone, both passing the blood-brain barrier. In the final aim we will establish an in vivo protocol of transplanting human iPSC-derived astrocytes into neonatal mice and induce their neuronal conversion at adult stages. This will provide an in vivo assay for human astrocyte conversion into neurons to explore the role of ferroptosis in this process.This pioneering approach of examining the role of ferroptosis in direct neuronal reprogramming will greatly profit from the SPP network on ferroptosis and provide a unique platform for exploring ferroptosis mechanisms in human cells.

在哺乳动物的大多数大脑区域中，因脑损伤或神经退行性疾病而丢失的神经元无法替代。我们开创了从局部神经胶质细胞直接进行神经元重编程的新方法，并确定铁死亡是这一过程中的主要障碍。在这里，我们的目标是进一步剖析当小鼠和人类来源的星形胶质细胞在体外转化为神经元时引发铁死亡所涉及的途径。我们将进一步探索与未折叠蛋白反应的可能联系，并分析在我们最近的星形胶质细胞和神经元线粒体蛋白质组中鉴定的神经元特异性抗氧化蛋白的作用。基于体外最有希望的结果，我们的目标是在体内小鼠大脑皮层刺伤后转化反应性星形胶质细胞时特别抑制这些途径，例如，通过过表达 FSP1、Gpx4、删除 Acsl4 或使用 CRISPRa 来激活神经元特异性线粒体蛋白的表达。这些基因操作将与药物治疗的效果进行比较，例如通过使用第三代利普司他汀或罗格列酮等抑制剂，两者均可通过血脑屏障。我们的最终目标是建立一种体内方案，将人 iPSC 衍生的星形胶质细胞移植到新生小鼠体内，并在成年阶段诱导其神经元转化。这将为人类星形胶质细胞转化为神经元提供一种体内检测方法，以探索铁死亡在此过程中的作用。这种检查铁死亡在直接神经元重编程中的作用的开创性方法将极大地受益于铁死亡的SPP网络，并为探索人类细胞铁死亡机制提供独特的平台。