Single Particle Characterization and Isolation of Extracellular Vesicles

细胞外囊泡的单颗粒表征和分离

• 批准号: RGPIN-2017-05974
• 负责人: Langlois, MarcAndré
• 金额: $ 1.89万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=681671
• 关键词: Single; Particle; Characterization; Isolation; Extracellular

Extracellular vesicles (EVs) are submicron-sized biological particles secreted by all forms of life such as cells, bacteria and yeast. EVs carry as cargo molecules from the cells from which they were released (e.g., proteins, lipids, nucleic acids). These can in turn exert biological effects in cells that uptake them such as inducing growth, proliferation, migration and even the secretion of soluble bioactive factors. EVs are released in instances of stress, damage and disease, but can also be released as a means of general communication between biological entities to accomplish a specific task. Proteins and markers at the surface of individual EVs guide them towards their intended target cells that will then engulf them. In complex environments such as bacterial biofilms and biological fluids, EVs from all the cells in the system are released together into the extracellular environment. Because EVs share nearly identical biophysical properties, current purification methods isolate EVs as a bulk. It is therefore nearly impossible to study EV diversity or isolate EVs emanating from specific cells in the system. As such, very little is known about EV diversity, subpopulations and how they impact the behavior of recipient cells in complex and heterogeneous systems. ******My research program aims to understand the function of EVs and how they alter cells in complex environments. To achieve this goal, I propose to:****** i) Establish a new technological analysis platform called NanoFlow that enables single particle characterization and isolation. *** i) Profile surface proteins and lipids, and characterize the diversity of EVs secreted in various complex environments and biological fluids. *** iii) Isolate specific EV populations by NanoFlow sorting, analyze their protein, lipid and nucleic acid content, and determine their biological effects on recipient cells.******The insight provided by detailed EV characterizations will help to better understand how EVs are used to communicate information between cells and coordinate biological effects. Furthermore, the specialized nature of the technology that will be developed by my research program will provide trainees with a unique set of translatable and marketable skills that will no doubt be useful for their future careers in academic research and industry.

细胞外囊泡（Extracellular vesicles，EV）是由所有生命形式如细胞、细菌和酵母分泌的亚微米级生物颗粒。EV携带来自释放它们的细胞的货物分子（例如，蛋白质、脂质、核酸）。这些可以反过来在摄取它们的细胞中发挥生物学作用，例如诱导生长、增殖、迁移，甚至分泌可溶性生物活性因子。EV在压力，损伤和疾病的情况下释放，但也可以作为生物实体之间的一般通信手段来完成特定任务。单个EV表面的蛋白质和标记物引导它们朝向其预期的靶细胞，然后吞噬它们。在复杂的环境中，如细菌生物膜和生物流体，来自系统中所有细胞的EV一起释放到细胞外环境中。由于EV具有几乎相同的生物物理特性，因此目前的纯化方法将EV作为整体分离。因此，几乎不可能研究EV多样性或分离系统中特定细胞发出的EV。因此，人们对EV多样性、亚群以及它们如何影响复杂和异质系统中受体细胞的行为知之甚少。* 我的研究项目旨在了解电动汽车的功能以及它们如何在复杂环境中改变细胞。为了实现这一目标，我建议：* i）建立一个名为NanoFlow的新技术分析平台，可以进行单颗粒表征和分离。* i）分析表面蛋白和脂质，并表征在各种复杂环境和生物流体中分泌的EV的多样性。 * iii）通过NanoFlow分选分离特定EV群体，分析其蛋白质、脂质和核酸含量，并确定其对受体细胞的生物学效应。详细的EV表征提供的洞察力将有助于更好地理解EV如何用于在细胞之间传递信息并协调生物效应。此外，我的研究项目将开发的技术的专业性质将为学员提供一套独特的可翻译和可销售的技能，这无疑将对他们未来的学术研究和工业生涯有用。