Intra- and intercellular functions of redoxins during neuroinflammation

可逆氧化性硫醇修饰是控制信号转导的翻译后修饰，使硫醇开关对细胞功能具有极其重要的意义。这些开关受到硫氧还蛋白家族的氧化还原蛋白-氧化还原酶的严格调控。在我们的提案中，我们将讨论这个蛋白质家族的几个仍然鲜为人知的方面。我们希望增加1)表征底物的数量并提高对ii)底物专一性的理解，iii)除了蛋白(In)激活之外的调控变化，例如易位或与其他修饰的相互作用，以及iv)氧化还蛋白的时空活性，特别是分泌的氧化还原蛋白的细胞外功能。根据我们在神经学和炎症方面的经验，我们将结合体外(细胞培养)、体外(原代细胞、器官切片培养)和体内模型(斑马鱼、小鼠)以及多发性硬化症患者的样本(脑片、血清、脑脊液)，协同分析氧化还蛋白在神经炎症中的作用。在我们的两个项目中，我们将研究神经胶质细胞再生和免疫调节过程中氧化还蛋白调节的过程及其潜在的分子机制。炎症诱导的脑损伤的再生，从而对神经功能缺陷的保护依赖于少突胶质前体细胞向损伤的迁移及其重新髓鞘形成的能力。我们假设谷氧还蛋白2诱导的特定状态的分化阻断(NG2-胶质细胞)在不同的炎症模式(自身免疫、创伤)中促进神经元的再生。为了分析分泌的氧化还蛋白的免疫调节机制，我们将研究参与神经炎症的不同类型的细胞，特别是微小细胞和星形胶质细胞的氧化还蛋白的分泌，并表征它们的细胞外底物和功能。基于初步数据，我们建议将分泌型氧化还蛋白作为疾病进展和免疫调节治疗有效性的新诊断工具，这将通过将我们的发现翻译到多发性硬化症患者来进行研究。除了氧化还蛋白去除氧化的硫醇修饰外，我们还将从机制和生理水平上研究这些修饰的形成，重点是潜在的硫醇氧化酶赖氨酸特异性去甲基酶1。总之，我们的两个项目首次全面研究了神经胶质细胞中的氧化还原蛋白功能。选择神经炎症的病理状态，重点是多发性硬化症，我们的建议将提供具有直接临床影响的翻译结果，以及对细胞内和细胞间信号的氧化还蛋白和氧化还原调节的基本机制和功能洞察。

Reversible oxidative thiol modifications are posttranslational modifications that control signal transduction making thiol switches of utmost significance for cellular functions. These switches are tightly regulated by redoxins - oxidoreductases of the thioredoxin family.In our proposal, we will address several aspects of this protein family that are still barely understood. We want to increase i) the number of characterized substrates and improve the understanding of ii) substrate specificity, iii) regulatory variety in addition to protein (in)activation, e.g. translocation or the interplay with other modifications, and the iv) spatio-temporal activity of redoxins, in particular extracellular functions of secreted redoxins. Based on our experiences in neurology and inflammation, we will synergistically analyze the functions of redoxins in neuroinflammation combining in vitro (cell cultures), ex vivo (primary cells, organotypic slice cultures), and in vivo models (zebrafish, mice) as well as samples from multiple sclerosis patients (brain slices, serum, cerebrospinal fluid). Within our two projects we will investigate redoxin-regulated processes and their underlying molecular mechanisms in glia cells during regeneration and immunomodulation. The regeneration from inflammation-induced brain damage and thereby the protection against neurological deficits depends on the migration of oligodendroglial progenitor cells towards lesions and their remyelinating capacity. We hypothesize that the identified glutaredoxin 2-induced differentiation block in a specific state (NG2-glia cells) enhances regeneration of neurons in different inflammation paradigms (autoimmunity, traumatic injury).To analyze the immunomodulatory mechanisms of secreted redoxins, we will investigate the secretion of redoxins by distinct cell types involved in neuroinflammation, in particular micro- and astroglia, and characterize their extracellular substrates and functions. Based on preliminary data, we propose secreted redoxins as new diagnostic tools for disease progression and effectiveness of immunomodulatory therapy which will be investigated by translating our findings to patients suffering from multiple sclerosis. In addition to the removal of oxidative thiol modifications by redoxins, we will also investigate the formation of these modifications on a mechanistic and physiologic level, focusing on the potential thiol oxidase lysine specific demethylase1. In summary, our two projects provide the first comprehensive investigation of redoxin functions in glial cells. Choosing the pathological condition of neuroinflammation, with emphasis on multiple sclerosis, our proposal will provide both translational results with direct clinical impact and fundamental mechanistic and functional insights into redoxins and redox regulation of intra- and intercellular signaling.