Role of extracellular vesicle and DJ-1 in intercellular communication

大脑的正确功能需要不同类型细胞之间复杂、快速和有效的细胞间通讯。神经细胞和神经胶质细胞都能产生细胞外小泡(EVS)，作为细胞间通讯的媒介。电动汽车是通过多条途径产生的异质膜结合舱，产生不同的类型和货物。它们与生理和病理的细胞间通讯过程有关，因为它们可以通过血脑屏障传输核酸、蛋白质、脂类以及与疾病相关的分子。最近的研究表明，它们作为生物标志物的潜力，因为在血液、尿液和脑脊液等生物液中发现了神经源性EV。在此背景下，DJ-1是一种特别有趣的蛋白质：它在氧化应激保护中发挥主要作用，它作为免疫细胞调节器存在于电动汽车中。此外，DJ-1基因突变可导致常染色体隐性遗传性帕金森病，其在帕金森病发病机制中的分子机制尚不清楚。本课题的目的是研究EVS和DJ-1在生理和病理细胞间通讯中的作用。我们将通过回答以下问题来实现这一目标：在没有DJ-1的情况下，EV种群如何变化，DJ-1货物对受体细胞的影响是什么？我们将使用对照或DJ-1基因敲除的SH-SY5Y细胞，以及野生型、帕金森病致病株和工程DJ-1突变体，以确定EVS定位的DJ-1在健康和由神经毒素或突触核蛋白原纤维治疗诱导的帕金森病细胞模型中的细胞间通信中的作用。通过流式细胞仪、RNAseq和蛋白质组学的研究，我们的结果将阐明DJ-1 Cargo在细胞间通讯中的作用，并为进一步研究EVS在神经退行性变中的作用铺平道路。

Correct function of the brain requires complex, rapid, and efficient intercellular communication between varied cell types. Both neuronal and glial cells can produce extracellular vesicles (EVs) as agents of intercellular communication. EVs are heterogenous membrane-bound compartments produced through multiple pathways, resulting in different types and cargo. They are implicated in both physiological and pathological intercellular communication processes, as they can transfer nucleic acids, proteins, and lipids as well as disease-related molecules through the blood-brain barrier. Recent studies have shown their potential as biomarkers, as neuronal derived EVs were identified in biofluids such as blood, urine and cerebrospinal fluid. Within this context, DJ-1 is a particularly interesting protein: it plays a major role in oxidative stress protection, it acts as immune cell modulator and it is present in EVs. Furthermore, mutations in DJ-1 cause autosomal recessive PD and the molecular mechanisms underlying DJ-1 contribution to PD pathogenesis are still unclear.The aim of this project is to characterise and understand the role of EVs and DJ-1 in physiological and pathological intercellular communication. We will achieve this goal by answering the following questions: how does EV population change in absence of DJ-1 and what is the effect of DJ-1 cargo on recipient cells? We will employ control or DJ-1 knockout SH-SY5Y cells, as well as wild-type, PD causative and engineered DJ-1 mutants to determine the role of EVs localised DJ-1 in intercellular communication in healthy and parkinsonian cell models induced by neurotoxin or -synuclein fibril treatment. Through study by flow cytometry, RNAseq, and proteomics our results will clarify the role of DJ-1 cargo in intercellular communication and pave the way for future studies on EVs role in neurodegeneration.