外泌体分离技术及其临床应用研究进展

doi:10.3724/SP.J.1123.2019.02007

摘要/Abstract

摘要： 外泌体是细胞通过胞吐过程分泌的一类粒径为30~200 nm的囊泡，其组成包括脂质双分子层以及其内部包裹的细胞来源的蛋白质、核糖核苷酸（RNA）和脱氧核糖核苷酸（DNA）等生物分子。作为一种细胞间交流的重要方式，外泌体在一系列生理和病理过程中起着至关重要的作用。由于体液环境复杂，加之自身体积小、密度低，外泌体的富集与分离对于其后续分析和功能研究至关重要。该文介绍了外泌体的研究策略、表征手段及生物学功能和临床应用研究进展，特别对外泌体的提取方法进行了详细介绍，并加以系统评述。

关键词: 外泌体, 临床应用, 提取方法, 综述

Abstract: Exosomes are vesicles secreted by many types of cells through exocytosis, and their sizes range from 30 to 200 nm. Exosomes consist of a lipid bilayer membrane, containing a number of bioactive molecules, e. g., proteins, ribose nucleic acid (RNA), and deoxyribo nucleic acid (DNA) derived from the cell of origin. As intercellular communication carriers, exosomes participate in many physiological and pathological processes. Because of the complexity of body fluids, as well as the small size and low density of exosomes, the isolation of exosomes is an essential and challenging step before subsequent analysis and functional studies. This review summarizes the advances in the analytical approaches, characterization methods, biological functions and clinical applications of exosomes, with particular emphasis on exosomes isolation techniques.

Key words: exosome, clinical application, isolation method, review

中图分类号:

O658

高方园, 焦丰龙, 张养军, 秦伟捷, 钱小红. 外泌体分离技术及其临床应用研究进展[J]. 色谱, 2019, 37(10): 1071-1083.

GAO Fangyuan, JIAO Fenglong, ZHANG Yangjun, QIN Weijie, QIAN Xiaohong. Advances in separation techniques for exosomes and their clinical applications[J]. Chinese Journal of Chromatography, 2019, 37(10): 1071-1083.

参考文献

[1] Pan B T, Johnstone R M. Cell, 1983, 33(3):967
[2] Johnstone R M, Adam M, Hammond J R, et al. J Biol Chem, 1987, 262(19):9412
[3] Keller S, Sanderson M P, Stoeck A, et al. Immunol Lett, 2006, 107(2):102
[4] Suchorska W M, Lach M S. Oncol Rep, 2016, 35(3):1237
[5] Shao H, Im H, Castro C M, et al. Chem Rev, 2018, 118(4):1917
[6] Llorente A, Skotland T, Sylvänne T, et al. BBA-Mol Cell Biol L, 2013, 1831(7):1302
[7] Purushothaman A, Bandari S K, Liu J, et al. J Biol Chem, 2016, 291(4):1652
[8] Urbanelli L, Magini A, Buratta S, et al. Genes, 2013, 4(2):152
[9] Meldolesi J. Curr Biol, 2018, 28(8):435
[10] Valadi H, Ekström K, Bossios A, et al. Nat cell biol, 2007, 9(6):654
[11] Vlassov A, Magdaleno S, Setterquist R, et al. BBA-Gen Subjects, 2012, 1820(7):940
[12] Stoorvogel W, Kleijmeer M J, Geuze H J, et al. Traffic, 2002, 3(5):321
[13] Vaidyanathan R, Naghibosadat M, Rauf S, et al. Anal Chem, 2014, 86(22):11125
[14] WangY Q, Sui Z G, Zhang L H, et al. Chinese Journal of Chromatography, 2016, 34(12):1131 翁叶靖, 随志刚, 张丽华, 等. 色谱, 2016, 34(12):1131
[15] Choi D S, Kim D K, Kim Y K, et al. Mass Spectrom Rev, 2015, 34(4):474
[16] Zabeo D, Cvjetkovic A, Lässer C, et al. J Extracell Vesicles, 2017, 6(1):1329476
[17] Yang J S, Lee J C, Byeon S K, et al. Anal Chem, 2017, 89(4):2488
[18] Bobrie A, Théry C. Biochem Soc Trans, 2013, 41(1):263
[19] Raposo G, Nijman H W, Stoorvogel W, et al. J Exp Med, 1996, 183(3):1161
[20] Van Niel G, Raposo G, Candalh C, et al. Gastroentero-logy, 2001, 121(2):337
[21] Momen-Heravi F, Balaj L, Alian S, et al. Front Physiol, 2012, 3:162
[22] An M, Lohse I, Tan Z, et al. J Proteome Res, 2017, 16(4):1763
[23] Kalra H, Adda C G, Liem M, et al. Proteomics, 2013, 13(22):3354
[24] Keller S, Ridinger J, Rupp A-K, et al. J Transl Med, 2011, 9(1):86
[25] Welton J L, Webber J P, Botos L-A, et al. J Extracell Vesicles, 2015, 4(1):27269
[26] Coumans F A, Brisson A R, Buzas E I, et al. Circ Res, 2017, 120(10):1632
[27] Cheruvanky A, Zhou H, Pisitkun T, et al. Am J Physiol-Renal, 2007, 292(5):F1657
[28] Abramowicz A, Widlak P, Pietrowska M. Mol BioSyst, 2016, 12(5):1407
[29] Zhao H, Yang L, Baddour J, et al. eLife, 2016, 5:e10250
[30] Tauro B J, Greening D W, Mathias R A, et al. Methods, 2012, 56(2):293
[31] Mathivanan S, Lim J W E, Tauro B J, et al. Mol Cell Proteomics, 2010, 9(2):197
[32] Koga K, Matsumoto K, Akiyoshi T, et al. Anticancer Res, 2005, 25(6A):3703
[33] Mizutani K, Terazawa R, Kameyama K, et al. Anticancer Res, 2014, 34(7):3419
[34] Echevarria J, Royo F, Pazos R, et al. Chembiochem, 2014, 15(11):1621
[35] Nakai W, Yoshida T, Diez D, et al. Sci Rep, 2016, 6:33935
[36] Kim J, Shin H, Kim J, et al. PloS One, 2015, 10(6):e0129760
[37] Zarovni N, Corrado A, Guazzi P, et al. Methods, 2015, 87:46
[38] Ko J, Bhagwat N, Black T, et al. Cancer Res, 2018, 78(13):3688
[39] Shin S, Han D, Park M C, et al. Sci Rep, 2017, 7(1):9907
[40] Woo H-K, Sunkara V, Park J, et al. ACS Nano, 2017, 11(2):1360
[41] Davies R T, Kim J, Jang S C, et al. Lab Chip, 2012, 12(24):5202
[42] Yasui T, Yanagida T, Ito S, et al. Sci Adv, 2017, 3(12):e1701133
[43] Lewis J M, Vyas A D, Qiu Y, et al. ACS Nano, 2018, 12(4):3311
[44] Ibsen S D, Wright J, Lewis J M, et al. ACS Nano, 2017, 11(7):6641
[45] Kang Y-T, Kim Y J, Bu J, et al. Nanoscale, 2017, 9(36):13495
[46] Reátegui E, Vos K E, Lai C P, et al. Nat Commun, 2018, 9(1):175
[47] Liu C, Guo J, Tian F, et al. ACS Nano, 2017, 11(7):6968
[48] Wu M, Ouyang Y, Wang Z, et al. Proc Natl Acad Sci U S A, 2017, 114(40):10584
[49] Wu X, Li L, Iliuk A, et al. J Proteome Res, 2018, 17(9):3308
[50] Wan Y, Cheng G, Liu X, et al. Nat Biomed Eng, 2017, 1(4):0058
[51] Gao F, Jiao F, Xia C, et al. Chem Sci, 2019, 10:1565
[52] Li P, Kaslan M, Lee S H, et al. Theranostics, 2017, 7(3):789
[53] Pisitkun T, Shen R F, Knepper M A. Proc Natl Acad Sci U S A, 2004, 101(36):13368
[54] Melo S A, Luecke L B, Kahlert C, et al. Nature, 2015, 523(7559):177
[55] Van Der Pol E, Hoekstra A, Sturk A, et al. J Thromb Haemostasis, 2010, 8(12):2596
[56] Tang Y-T, Huang Y-Y, Zheng L, et al. Int J Mol Med, 2017, 40(3):834
[57] Théry C, Amigorena S, Raposo G, et al. Curr Protoc Cell Biol, 2006:3.22.1
[58] Comelli L, Rocchiccioli S, Smirni S, et al. Mol Bio Syst, 2014, 10(5):1146
[59] Tian Y, Ma L, Gong M, et al. ACS Nano, 2018, 12(1):671
[60] Mathivanan S, Ji H, Simpson R J. J Proteomics, 2010, 73(10):1907
[61] Barry O P, Praticò D, Savani R C, et al. J Clin Invest, 1998, 102(1):136
[62] Yáñez-Mó M, Siljander P R-M, Andreu Z, et al. J Extracell Vesicles, 2015, 4(1):27066
[63] He M, Qin H, Poon T C, et al. Carcinogenesis, 2015, 36(9):1008
[64] Iero M, Valenti R, Huber V, et al. Cell Death Differ, 2008, 15(1):80
[65] Zomer A, Maynard C, Verweij F, et al. Cell, 2015, 161(5):1046
[66] Hoshino A, Costa-Silva B, Shen T L, et al. Nature, 2015, 527(7578):329
[67] Costa-Silva B, Aiello N M, Ocean A J, et al. Nat Cell Biol, 2015, 17(6):816
[68] Peinado H, Aleckovic M, Lavotshkin S, et al. Nat Med, 2012, 18(6):883
[69] Maji S, Chaudhary P, Akopova I, et al. Mol Cancer Res, 2017, 15(1):93
[70] Qu L, Ding J, Chen C, et al. Cancer Cell, 2016, 29(5):653
[71] Yeung C L A, Co N-N, Tsuruga T, et al. Nat Commun, 2016, 7:11150
[72] Mikamori M, Yamada D, Eguchi H, et al. Sci Rep, 2017, 7:42339
[73] Qin X, Yu S, Xu X, et al. Oncotarget, 2017, 8(26):42125
[74] Chen G, Huang A C, Zhang W, et al. Nature, 2018, 560:382
[75] Chalmin F, Ladoire S, Mignot G, et al. J Clin Invest, 2010, 120(2):457
[76] Wang X, Luo G, Zhang K, et al. Cancer Res, 2018, 78(16):4586
[77] Arscott W T, Tandle A T, Zhao S, et al. Transl Oncol, 2013, 6(6):638
[78] Kumar Jella K, Rani S, O'driscoll L, et al. Radiat Res, 2014, 181(2):138
[79] Record M, Subra C, Silvente-Poirot S, et al. Biochem Pharmacol, 2011, 81(10):1171
[80] Zitvogel L, Regnault A, Lozier A, et al. Nat Med, 1998, 4:594
[81] Munich S, Sobovujanovic A, Buchser W J, et al. Oncoimmunology, 2012, 1(7):1074
[82] Sanchita B, Schorey J S. J Biol Chem, 2007, 282(35):25779
[83] Skokos D, Panse S, Le, Villa I, et al. J Immunol, 2001, 166(2):868
[84] Vlist E J V D, Arkesteijn G J A, Lest C H a V D, et al. J Extracell Vesicles, 2012, 1(1):18364
[85] Arslan F, Lai R C, Smeets M B, et al. Stem Cell Res, 2013, 10(3):301
[86] Bruno S, Collino F, Deregibus M C, et al. Stem Cells Dev, 2012, 22(5):758
[87] Rahman M J, Regn D, Bashratyan R, et al. Diabetes, 2014, 63(3):1008
[88] Zhu W, Huang L, Li Y, et al. Cancer Lett, 2012, 315(1):28
[89] Kalluri R. J Clin Invest, 2016, 126(4):1208
[90] Gallo A, Tandon M, Alevizos I, et al. PloS One, 2012, 7(3):e30679
[91] Wang N, Wang L, Yang Y, et al. Biochem Biophys Res Commun, 2017, 493(3):1322
[92] Liu Q, Yu Z, Yuan S, et al. Oncotarget, 2017, 8(8):13048
[93] Lugli G, Cohen A M, Bennett D A, et al. PloS One, 2015, 10(10):e0139233
[94] Taylor D D, Gercel-Taylor C. Gynecol Oncol, 2008, 110(1):13
[95] Shi J. J Clin Med, 2016, 5(4):42
[96] Tanaka Y, Kamohara H, Kinoshita K, et al. Cancer, 2013, 119(6):1159
[97] Tsukamoto M, Iinuma H, Yagi T, et al. Oncology, 2017, 92(6):360
[98] Nilsson J, Skog J, Nordstrand A, et al. Br J Cancer, 2009, 100(10):1603
[99] Rodríguez M, Bajo-Santos C, Hessvik N P, et al. Mol Cancer, 2017, 16(1):156
[100] Khurana R, Ranches G, Schafferer S, et al. RNA, 2017, 23(2):142
[101] Skotland T, Ekroos K, Kauhanen D, et al. Eur J Cancer, 2017, 70:122
[102] Michael A, Bajracharya S D, Yuen P S, et al. Oral Dis, 2010, 16(1):34
[103] Wu C-X, Liu Z-F. J Invest Dermatol, 2018, 138(1):89
[104] Liu C-G, Song J, Zhang Y-Q, et al. Mol Med Rep, 2014, 10(5):2395
[105] Morse M A, Garst J, Osada T, et al. J Transl Med, 2005, 3(1):9
[106] Escudier B, Dorval T, Chaput N, et al. J Transl Med, 2005, 3(1):10
[107] Hood J L, San R S, Wickline S A. Cancer Res, 2011, 71(11):3792
[108] Pascucci L, Coccè V, Bonomi A, et al. J Control Release, 2014, 192:262
[109] Tian Y, Li S, Song J, et al. Biomaterials, 2014, 35(7):2383
[110] Haney M J, Klyachko N L, Zhao Y, et al. J Control Release, 2015, 207:18
[111] Yang J, Zhang X, Chen X, et al. Mol Ther-Nucl Acids, 2017, 7:278
[112] Zhuang X, Xiang X, Grizzle W, et al. Mol Ther, 2011, 19(10):1769