Yasui Takao, Paisrisarn Piyawan, Yanagida Takeshi, Konakade Yuki, Nakamura Yuta, Nagashima Kazuki, Musa Marina, Thiodorus Ivan Adiyasa, Takahashi Hiromi, Naganawa Tsuyoshi, Shimada Taisuke, Kaji Noritada, Ochiya Takahiro, Kawai Tomoji, Baba Yoshinobu
Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan; Japan Science and Technology Agency (JST), PRESTO, 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012, Japan; Institute of Nano-Life-Systems, Institutes of Innovation for Future Society, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Biosens Bioelectron. 2021 Dec 15;194:113589. doi: 10.1016/j.bios.2021.113589. Epub 2021 Aug 27.
Extracellular vesicles (EVs) have shown promising features as biomarkers for early cancer diagnoses. The outer layer of cancer cell-derived EVs consists of organotropic metastasis-induced membrane proteins and specifically enriched proteoglycans, and these molecular compositions determine EV surface charge. Although many efforts have been devoted to investigating the correlation between EV subsets obtained through density-, size-, and immunoaffinity-based captures and expressed membrane proteins, understanding the correlation between EV subsets obtained through surface charge-based capture and expressed membrane proteins is lacking. Here, we propose a methodology to profile membrane proteins of EV subsets obtained through surface charge-based capture. Nanowire-induced charge-based capture of EVs and in-situ profiling of EV membrane proteins are the two key methodology points. The oxide nanowires allowed EVs to be obtained through surface charge-based capture due to the diverse isoelectric points of the oxides and the large surface-to-volume ratios of the nanowire structures. And, with the ZnO nanowire device, whose use does not require any purification and concentration processes, we demonstrated the correlation between negatively-charged EV subsets and expressed membrane proteins derived from each cell. Furthermore, we determined that a colon cancer related membrane protein was overexpressed on negatively charged surface EVs derived from colon cancer cells.
细胞外囊泡（EVs）作为早期癌症诊断的生物标志物已展现出有前景的特征。癌细胞衍生的EVs外层由器官趋向性转移诱导的膜蛋白和特异性富集的蛋白聚糖组成，这些分子组成决定了EV的表面电荷。尽管已经做出很多努力来研究通过基于密度、大小和免疫亲和的捕获方法获得的EV亚群与表达的膜蛋白之间的相关性，但对于通过基于表面电荷的捕获方法获得的EV亚群与表达的膜蛋白之间的相关性仍缺乏了解。在此，我们提出一种方法来分析通过基于表面电荷的捕获方法获得的EV亚群的膜蛋白。基于纳米线诱导电荷的EV捕获和EV膜蛋白的原位分析是两个关键的方法要点。由于氧化物的等电点多样且纳米线结构的表面积与体积比大，氧化物纳米线使得能够通过基于表面电荷的捕获方法获得EVs。并且，使用无需任何纯化和浓缩过程的ZnO纳米线装置，我们证明了带负电荷的EV亚群与源自每个细胞的表达膜蛋白之间的相关性。此外，我们确定一种结肠癌相关膜蛋白在源自结肠癌细胞的带负电荷的表面EVs上过度表达。
