Elaboration of a Bio-Plex platform for the characterization of Exosomes

所有类型的细胞都会释放细胞外小泡(EV)和外切体。外体是异质囊泡家族的一部分，在细胞间通讯中起着至关重要的作用。它们可以被邻近或远处的细胞选择性地摄取来运送它们的货物，并对这些细胞的生理或病理功能做出贡献。EVS/exosome的研究领域正在迅速扩大，研究其生理和病理功能的研究呈指数级增长。它们可能导致各种慢性疾病，包括“长期肺炎”症状或阿尔茨海默病。此外，EVS/exosome作为一种新型的纳米药物传递工具也很有前景。到目前为止，该领域的局限性是缺乏采集和鉴定EVS/exosome的金标准方法。它们必须用几种互补的工艺来表征它们的大小、形态、电荷等。然而，考虑到其生物发生的复杂性，只有对几个特定标记物的免疫检测才能验证外切体在其他纳米尺寸的囊泡中的存在。事实上，分离后对其标志物的免疫检测大多局限于每个样本一到两个分析物，并且耗时和样本消耗。这项提议的原创性是开发一个ExoBio-Plex平台，旨在以非常有限的样本量同时覆盖14个EVS/Exosomal标记。此外，为了更好地了解EVS/外显子在新冠肺炎和一些慢性疾病中的作用，我们将把这个平台与其他Bio-Plex平台结合起来，例如Bio-Plex MultiplexPro SARS-CoV-2检测，或带有48-Plex的人类细胞因子筛选小组，或与阿尔茨海默病相关标记物的分别开发的ExoCovid-19 Bio-Plex，Exo-Cytokines-Bio-Plex，以及ExoAlz-Bio-Plex平台。这些平台将在来自不同细胞类型的不同生物液和细胞培养液上进行验证。它们将加快对EVS/exosome作用的基础和应用研究，将为其在其他慢性病中的应用开辟新的途径。

Extracellular vesicles (EVs) and exosomes are released by all cell types. A part of a heterogeneous family of vesicles, exosomes are vital in intercellular communication. They can be selectively taken up by neighboring or distant cells to deliver their cargo and contribute to the physiological or pathological functions of these cells. The field of EVs/Exosomes is expanding very rapidly, with an exponential increase in studies investigating their physiological and pathological functions. They can contribute to various chronic diseases including in ``Long COVID``symptoms or in Alzheimer's disease. Moreover, EVs/Exosomes could also be promising as novel nano-drug delivery tools.To date, the limitation in this field is the lack of the gold-standard method to harvest and to identify EVs/exosomes. They have to be characterized with several complementary technics for their size, morphology, charges..etc. However, considering the complexity of their biogenesis only the immune-detection of several specific markers could validate the presence of exosomes among other nanosized vesicles. Actually, the immunodetection of their markers following isolation is mostly limited to one or two analytes per sample and is time and sample-consuming. The originality of this proposal is to develop an ExoBio-Plex platform, designed to cover 14 EVs/Exosomal markers simultaneously with a very limited volume sample. Moreover, to better understand the role of EVs/exosomes in COVID-19 and in some chronic diseases, we will combine this platform to other Bio-Plex platforms such as the Bio-Plex MultiplexPro SARS-CoV-2 Assays, or Human Cytokine Screening Panel with 48-Plex, or with Alzheimer's related markers to develop respectively, the ExoCovid-19 Bio-Plex, the Exo-Cytokines-Bio-Plex, and the ExoAlz-Bio-Plex platforms. These platforms will be validated on different biofluids and cell culture media from different cell types. They will accelerate the fundamental and applied research on the role of EVs/Exosomes, will open new avenues for their application to other chronic diseases.