尿液细胞外囊泡的单颗粒水平多参数定量表征

刘海生 袁文理 陈永钰 陈晨 田野 颜晓梅

引用本文: 刘海生,  袁文理,  陈永钰,  陈晨,  田野,  颜晓梅. 尿液细胞外囊泡的单颗粒水平多参数定量表征[J]. 分析化学, 2020, 48(10): 1305-1314. doi: 10.19756/j.issn.0253-3820.201443 shu
Citation:  LIU Hai-Sheng,  YUAN Wen-Li,  CHEN Yong-Yu,  CHEN Chen,  TIAN Ye,  YAN Xiao-Mei. Multi-parameter and Quantitative Characterization of Urinary Extracellular Vesicles at Single-Particle Level[J]. Chinese Journal of Analytical Chemistry, 2020, 48(10): 1305-1314. doi: 10.19756/j.issn.0253-3820.201443 shu

尿液细胞外囊泡的单颗粒水平多参数定量表征

  • 基金项目:

    本文系国家自然科学基金项目(Nos.21934004,21627811)和国家基础科学人才培养基金项目(No.J1310024)资助

摘要: 尿液来源细胞外囊泡(Urinary extracellular vesicles,uEVs)作为新一类液体活检标志物,在疾病诊断、治疗监控领域具有广阔的应用前景。然而,由于缺乏相应的表征技术,研究者对于纳米尺度uEVs的粒径分布、颗粒浓度以及生化表型等均缺乏了解。EVs具有高度个体差异性和多样性,因此迫切需要发展高通量、多参数的单颗粒检测技术。基于本研究组前期工作中结合瑞利散射和鞘流单分子荧光检测技术研制的纳米流式检测装置(Nano-flow cytometer,nFCM),本研究对超速离心法纯化得到的uEVs的纯度、浓度、粒径、DNA、RNA和蛋白标志物在单颗粒水平进行多参数定量表征,其中EVs的检测限低至40 nm。结果表明,uEVs的纯度高达92%,10位健康受试者uEVs的颗粒浓度主要分布在3.0×109~8.9×1010个/mL之间,粒径主要分布在40~120 nm之间。DNA和RNA主要位于较大粒径的uEVs的内部,且颗粒之间的丰度存在很大的异质性。此外,使用nFCM对蛋白标志物CD9、CD63、CD81和CD24的阳性比率进行了测定。纳米流式检测技术可为uEVs的生化性质研究和临床应用提供重要的技术支撑。

English


    1. [1]

      Cocozza F, Grisard E, Martin-Jaular L, Mathieu M, Thery C. Cell, 2020, 182(1):262

    2. [2]

      Mathieu M, Martin-Jaular L, Lavieu G, Thery C. Nat. Cell Biol., 2019, 21(1):9-17

    3. [3]

      Shah R, Patel T, Freedman J E. New Engl. J. Med., 2018, 379(10):958-966

    4. [4]

      Xu R, Rai A, Chen M S, Suwakulsiri W, Greening D W, Simpson R J. Nat. Rev. Clin. Oncol., 2018, 15(10):617-638

    5. [5]

      Thongboonkerd V, Mcleish K R, Arthur J M, Klein J B. Kidney Int., 2002, 62(4):1461-1469

    6. [6]

      Pisitkun T, Shen R F, Knepper M A. Proc. Natl. Acad. Sci. USA, 2004, 101(36):13368-13373

    7. [7]

      Turco A E, Lam W, Rule A D, Denic A, Lieske J C, Miller V M, Larson J J, Kremers W K, Jayachandran M. J. Extracell. Vesicles, 2016, 5:29642

    8. [8]

      Vermassen T, D'Herde K, Jacobus D, Van Praet C, Poelaert F, Lumen N, Callewaert N, Decaestecker K, Villeirs G, Hoebeke P, Van Belle S, Rottey S, Delanghe J. J. Clin. Pathol., 2017, 70(10):838-846

    9. [9]

      Lee J Y, McKinney K Q, Pavlopoulos A J, Niu M, Kang J W, Oh J W, Kim K P, Hwang S. Mol. Cells, 2018, 41(3):179-187

    10. [10]

      Lozano-Ramos S I, Bancu I, Carreras-Planella L, Monguio-Tortajada M, Canas L, Juega J, Bonet J, Armengol M P, Lauzurica R, Borras F E. Bmc Nephrol., 2018, 19:189

    11. [11]

      Stokman M F, Bijnsdorp I V, Schelfhorst T, Pham T V, Piersma S R, Knol J C, Giles R H, Bongers E M H F, Knoers N V A M, Lilien M R, Jimenez C R, Renkema K Y. J. Proteomics, 2019, 192:27-36

    12. [12]

      Woo H K, Park J, Ku J Y, Lee C H, Sunkara V, Ha H K, Cho Y K. Lab Chip, 2019, 19(1):87-97

    13. [13]

      Sun I O, Lerman L O. Diagnostics, 2020, 10(5):311

    14. [14]

      Zabeo D, Cvjetkovic A, Lasser C, Schorb M, Lotvall J, Hoog J L. J. Extracell. Vesicles, 2017, 6(1):1329476

    15. [15]

      Shao H, Im H, Castro C M, Breakefield X, Weissleder R, Lee H. Chem. Rev., 2018, 118(4):1917-1950

    16. [16]

      Sitar S, Kejzar A, Pahovnik D, Kogej K, Tusek-Znidaric M, Lenassi M, Zagar E. Anal. Chem., 2015, 87(18):9225-9233

    17. [17]

      Tian Y, Ma L, Gong M F, Su G Q, Zhu S B, Zhang W Q, Wang S, Li Z B, Chen C X, Li L H, Wu L N, Yan X M. ACS Nano, 2018, 12(1):671-680

    18. [18]

      Akers J C, Ramakrishnan V, Nolan J P, Duggan E, Fu C C, Hochberg F H, Chen C C, Carter B S. PLoS One, 2016, 11(2):e0149866

    19. [19]

      van der Pol E, Sturk A, van Leeuwen T, Nieuwland R, Coumans F. J. Thromb. Haemost., 2018, 16(6):1236-1245

    20. [20]

      Kowal J, Arras G, Colombo M, Jouve M, Morath J P, Primdal-Bengtson B, Dingli F, Loew D, Tkach M, Thery C. Proc. Natl. Acad. Sci. USA, 2016, 113(8):E968-E977

    21. [21]

      Zhang H Y, Freitas D, Kim H S, Fabijanic K, Li Z, Chen H Y, Mark M T, Molina H, Martin A B, Bojmar L, Fang J, Rampersaud S, Hoshino A, Matei I, Kenific C M, Nakajima M, Mutvei A P, Sansone P, Buehring W, Wang H J, Jimenez J P, Cohen-Gould L, Paknejad N, Brendel M, Manova-Todorova K, Magalhaes A, Ferreira J A, Osorio H, Silva A M, Massey A, Cubillos-Ruiz J R, Galletti G, Giannakakou P, Cuervo A M, Blenis J, Schwartz R, Brady M S, Peinado H, Bromberg J, Matsui H, Reis C A, Lyden D. Nat. Cell Biol., 2018, 20(3):332-343

    22. [22]

      Choi D, Montermini L, Jeong H, Sharma S, Meehan B, Rak J. ACS Nano, 2019, 13(9):10499-10511

    23. [23]

      Mateescu B, Kowal E J, van Balkom B W, Bartel S, Bhattacharyya S N, Buzas E I, Buck A H, de Candia P, Chow F W, Das S, Driedonks T A, Fernandez-Messina L, Haderk F, Hill A F, Jones J C, Van Keuren-Jensen K R, Lai C P, Lasser C, Liegro I D, Lunavat T R, Lorenowicz M J, Maas S L, Mager I, Mittelbrunn M, Momma S, Mukherjee K, Nawaz M, Pegtel D M, Pfaffl M W, Schiffelers R M, Tahara H, Thery C, Tosar J P, Wauben M H, Witwer K W, Nolte-'t Hoen E N. J. Extracell. Vesicles, 2017, 6(1):1286095

    24. [24]

      Zhu S B, Ma L, Wang S, Chen C X, Zhang W Q, Yang L L, Hang W, Nolan J P, Wu L N, Yan X M. ACS Nano, 2014, 8(10):10998-11006

    25. [25]

      Ma L, Zhu S B, Tian Y, Zhang W Q, Wang S, Chen C X, Wu L N, Yan X M. Angew. Chem. Int. Edit., 2016, 55(35):10239-10243

    26. [26]

      Yang L L, Zhu S B, Hang W, Wu L N, Yan X M. Anal. Chem., 2009, 81(7):2555-2563

    27. [27]

      Merchant M L, Rood I M, Deegens J K J, Klein J B. Nat. Rev. Nephrol., 2017, 13(12):731-749

    28. [28]

      Oeyen E, Van Mol K, Baggerman G, Willems H, Boonen K, Rolfo C, Pauwels P, Jacobs A, Schildermans K, Cho W C, Mertens I. J. Extracell. Vesicles, 2018, 7(1):1490143

    29. [29]

      Hogan M C, Johnson K L, Zenka R M, Charlesworth M C, Madden B J, Mahoney D W, Oberg A L, Huang B Q, Leontovich A A, Nesbitt L L, Bakeberg J L, McCormick D J, Bergen H R, Ward C J. Kidney Int., 2013, 85(5):1225-1237

    30. [30]

      Tian Y, Gong M F, Hu Y Y, Liu H S, Zhang W Q, Zhang M M, Hu X X, Aubert D, Zhu S B, Wu L N, Yan X M. J. Extracell. Vesicles, 2020, 9(1):1697028

    31. [31]

      Pospichalova V, Svoboda J, Dave Z, Kotrbova A, Kaiser K, Klemova D, Ilkovics L, Hampl A, Crha I, Jandakova E, Minar L, Weinberger V, Bryja V. J. Extracell. Vesicles, 2015, 4:25530

    32. [32]

      Oeyen E, Willems H,'T Kindt R, Sandra K, Boonen K, Hoekx L, De Wachter S, Ameye F, Mertens I. J. Extracell. Vesicles, 2019, 8(1):1676035

    33. [33]

      Musante L, Bontha S V, La Salvia S, Fernandez-Pineros A, Lannigan J, Le T H, Mas V, Erdbrugger U. Sci. Rep., 2020, 10:3701

    34. [34]

      Tataruch-Weinert D, Musante L, Kretz O, Holthofer H. J. Extracell. Vesicles, 2016, 5:30281

    35. [35]

      Ku A, Ravi N, Yang M J, Evander M, Laurell T, Lilja H, Ceder Y. PLoS One, 2019, 14(5):e0217507

    36. [36]

      Park S, Lee K, Park I B, Kim N H, Cho S, Rhee W J, Oh Y, Choi J, Nam S, Lee D H. Diabetes Res. Clin. Pr., 2020, 160:108010

    37. [37]

      McKiernan J, Donovan M J, O'Neill V, Bentink S, Noerholm M, Belzer S, Skog J, Kattan M W, Partin A, Andriole G, Brown G, Wei J T, Thompson I M, Carroll P. Jama Oncol., 2016, 2(7):882-889

    38. [38]

      Campos-Silva C, Suarez H, Jara-Acevedo R, Linares-Espinos E, Martinez-Pineiro L, Yanez-Mo M, Vales-Gomez M. Sci. Rep., 2019, 9:2042

    39. [39]

      Keller S, Rupp C, Stoeck A, Runz S, Fogel M, Lugert S, Hager H D, Abdel-Bakky M S, Gutwein P, Altevogt P. Kidney Int., 2007, 72(9):1095-1102

  • 加载中
计量
  • PDF下载量:  10
  • 文章访问数:  1160
  • HTML全文浏览量:  93
文章相关
  • 收稿日期:  2020-07-29
  • 修回日期:  2020-08-19
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

/

返回文章