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

构建用于外泌体表征的 Bio-Plex 平台

• 批准号: 570686-2021
• 负责人: RAMASSAMY, Charles
• 金额: $ 13.83万
• 依托单位: Institut national de la recherche scientifique
• 依托单位国家: 加拿大
• 项目类别: Alliance Grants
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=720637
• 关键词: Elaboration; Bio; Plex; platform; characterization

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.

细胞外囊泡（EV）和外泌体由所有细胞类型释放。作为囊泡异质家族的一部分，外泌体在细胞间通讯中至关重要。它们可以被邻近或远处的细胞选择性地摄取，以递送它们的货物，并有助于这些细胞的生理或病理功能。EV/外泌体领域正在迅速扩大，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COVID”症状或阿尔茨海默病。此外，EVs/Exosomes也有望成为新型的纳米药物载体，但目前该领域的局限性在于缺乏一种金标准的方法来收集和鉴定EVs/Exosomes。它们必须用几种互补的技术来表征它们的大小，形态，电荷。然而，考虑到它们生物发生的复杂性，只有几种特异性标志物的免疫检测才能验证外泌体在其他纳米尺寸囊泡中的存在。实际上，在分离后对其标志物的免疫检测大多限于每个样品的一种或两种分析物，并且是耗时和样品消耗的。该提案的独创性在于开发一个ExoBio-DNA平台，旨在用非常有限的样品体积同时覆盖14个EV/外泌体标记物。 此外，为了更好地了解EV/exosomes在COVID-19和一些慢性疾病中的作用，我们将联合收割机将该平台与其他生物诊断平台结合，例如Bio-Cumulative MultiplexPro SARS-CoV-2 Assays，或使用48-kDa或阿尔茨海默氏症相关标志物的人类细胞因子筛查面板，以分别开发ExoCovid-19生物诊断平台，Exo-Cytokines-Bio-Tumor和ExoAlz-Bio-Tumor平台。这些平台将在来自不同细胞类型的不同生物液体和细胞培养基上进行验证。它们将加速EV/Exosomes作用的基础和应用研究，为它们在其他慢性疾病中的应用开辟新的途径。