i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.
IPATIMUP - Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Porto, Portugal.
Methods Mol Biol. 2023;2619:125-139. doi: 10.1007/978-1-0716-2946-8_10.
Extracellular vesicles (EVs) have emerged as a central mechanism of intercellular communication in physiology and disease. EVs participate in paracrine exchange of nucleic acids as well as lipids, proteins, and glycans to elicit a complex biological response in target cells. Proteoglycans (PGs) are widely expressed in EV-producing cells and are sorted to the membrane of secreted EVs to participate in some of the key processes in EV-mediated signaling. Most notably, PGs mainly of the heparan sulfate (HS) type are involved in EV biogenesis and cellular uptake of EVs through endocytic processes. EV-associated PGs may serve as short- and long-range chaperones of signaling molecules with potential implications for intercellular information exchange, most importantly in cancer development. This motivates the development of approaches targeting EV-HSPG interactions as a strategy in cancer treatment. Moreover, the importance of PG remodeling and alterations in their expression in cancer, together with the fact that EVs mimic their cell or tissue of origin, point at an important role of EV-associated PGs as disease biomarkers. Here, we provide methodological insights into the analysis of EV-PGs isolated from cell cultures as well as patient plasma liquid biopsy.
细胞外囊泡 (EVs) 已成为生理和疾病中细胞间通讯的核心机制。EVs 通过旁分泌交换核酸以及脂质、蛋白质和聚糖，在靶细胞中引发复杂的生物学反应。糖胺聚糖 (PGs) 在产生 EV 的细胞中广泛表达，并被分拣到分泌的 EV 膜上，以参与 EV 介导的信号转导中的一些关键过程。值得注意的是，主要是硫酸乙酰肝素 (HS) 型的 PGs 参与 EV 的生物发生和通过内吞作用摄取 EV。EV 相关的 PGs 可能作为信号分子的短程和长程伴侣，对细胞间信息交换具有潜在意义，在癌症发展中尤为重要。这促使人们开发针对 EV-HSPG 相互作用的方法作为癌症治疗的一种策略。此外，PG 重塑及其在癌症中表达的改变，以及 EV 模拟其细胞或组织来源的事实，表明 EV 相关 PGs 作为疾病生物标志物的重要作用。在这里，我们提供了从细胞培养物和患者血浆液体活检中分离 EV-PGs 的分析方法学见解。
