Highly Selective Purification of Plasma Extracellular Vesicles Using Titanium Dioxide Microparticles for Depicting the Metabolic Signatures of Diabetic Retinopathy
Highly Selective Purification of Plasma Extracellular Vesicles Using Titanium Dioxide Microparticles for Depicting the Metabolic Signatures of Diabetic Retinopathy
复制标题
使用二氧化钛微粒高度选择性纯化血浆细胞外囊泡以描绘糖尿病视网膜病变的代谢特征
DOI：
10.1021/acs.analchem.1c05378
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发表时间：
2022-10-05
影响因子：
7.4
通讯作者：
Hu，Liang
中科院分区：
文献类型：
--
作者：
Pan，Youjin;Chen，Tucan;Hu，Liang
具有规律波动的细胞外囊泡（EV）货物有可能为临床诊断和预后提供化学预测因子。血浆样本是循环EV最重要的来源之一，但血浆EV分离的技术障碍和成本消耗仍然限制了其在疾病诊断和生物标志物发现中的应用。在这里，我们引入了一种易于使用的策略，可以选择性地从人血浆中纯化小型 EV（sEV）并检测其代谢变化。由于粗糙的 TiO2 壳的磷脂亲和力、增强的结合位点和尺寸排阻效应，具有粗糙岛状表面的 Fe3O4@TiO2 微珠已被证明能够进行有效且可逆的 sEV 捕获。与超速离心方法相比，所提出的系统还可以将分离过程从数小时缩短至20分钟，并从100μL血浆中产生约108sEV颗粒。最终研究了进行性糖尿病视网膜病变受试者中 sEV 的代谢组变化，观察到一系列水平升高的代谢物，并提示这些 sEV 化学物质在糖尿病视网膜病变发病和进展中的潜在作用。这种可扩展且灵活的 EV 捕获系统可以被视为促进基于血浆的液体活检的有效分析工具。
Extracellular vesicle (EV) cargos with regular fluctuations hold the potential for providing chemical predictors toward clinical diagnosis and prognosis. A plasma sample is one of the most important sources of circulating EVs, yet the technical barrier and cost consumption in plasma-EV isolation still limit its application in disease diagnosis and biomarker discovery. Here, we introduced an easy-to-use strategy that allows selectively purifying small EVs (sEVs) from human plasma and detecting their metabolic alternations. Fe3O4@TiO2microbeads with a rough island-shaped surface have proven the capability of performing efficient and reversible sEV capture owing to the phospholipid affinity, enhanced binding sites, and size-exclusion-like effect of the rough TiO2shell. The proposed system can also shorten the separation procedure from hours to 20 min when compared with the ultracentrifugation method and yield approximately 108sEV particles from 100 μL of plasma. Metabolome variations of sEVs among progressive diabetic retinopathy subjects were finally studied, observing a cluster of metabolites with elevated levels and suggesting potential roles of these sEV chemicals in diabetic retinopathy onset and progression. Such a scalable and flexible EV capture system can be seen as an effective analytical tool for facilitating plasma-based liquid biopsies.