Emerging techniques in the isolation and characterization of extracellular vesicles and their roles in cancer diagnostics and prognostics.

Extracellular vesicles (EVs) are cell-derived nanovesicles, present in almost all types of body fluids, which play an important role in intercellular communication and are involved in the transport of biological signals for regulating diverse cellular functions. Due to the increasing clinical interest in the role of EVs in tumor promotion, various techniques for their isolation, detection, and characterization are being developed. In this review, we present an overview of the current EV isolation and characterization methods in addition to their applications and limitations. Furthermore, EVs as the potential emerging biomarkers in cancer management and their clinical implementation are briefly discussed.

[1]  Maire Peters,et al.  Comparison of serum exosome isolation methods for microRNA profiling. , 2014, Clinical biochemistry.

[2]  M. Kesimer,et al.  Nanoparticle analysis of circulating cell-derived vesicles in ovarian cancer patients. , 2012, Analytical biochemistry.

[3]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[4]  S. Takayama,et al.  High-yield isolation of extracellular vesicles using aqueous two-phase system , 2015, Scientific Reports.

[5]  W. Ding,et al.  Intercellular Communication by Exosome-Derived microRNAs in Cancer , 2013, International journal of molecular sciences.

[6]  Romaric Lacroix,et al.  High-Sensitivity Flow Cytometry Provides Access to Standardized Measurement of Small-Size Microparticles—Brief Report , 2012, Arteriosclerosis, thrombosis, and vascular biology.

[7]  Zhiyuan Hu,et al.  Label-Free Quantitative Detection of Tumor-Derived Exosomes through Surface Plasmon Resonance Imaging , 2014, Analytical chemistry.

[8]  E. Messing,et al.  Bladder cancer exosomes contain EDIL-3/Del1 and facilitate cancer progression. , 2014, The Journal of urology.

[9]  C. Melief,et al.  B lymphocytes secrete antigen-presenting vesicles , 1996, The Journal of experimental medicine.

[10]  N. Kosaka,et al.  Exosomes from bone marrow mesenchymal stem cells contain a microRNA that promotes dormancy in metastatic breast cancer cells , 2014, Science Signaling.

[11]  Diane M Simeone,et al.  Microfluidic device (ExoChip) for on-chip isolation, quantification and characterization of circulating exosomes. , 2014, Lab on a chip.

[12]  D. Tang,et al.  Regulation of the Tumor Suppressor PTEN through Exosomes: A Diagnostic Potential for Prostate Cancer , 2013, PloS one.

[13]  Laurence Zitvogel,et al.  Eradication of established murine tumors using a novel cell-free vaccine: dendritic cell derived exosomes , 1998, Nature Medicine.

[14]  Jacco van Rheenen,et al.  In Vivo Imaging Reveals Extracellular Vesicle-Mediated Phenocopying of Metastatic Behavior , 2015, Cell.

[15]  M Kersaudy-Kerhoas,et al.  Exosome isolation: a microfluidic road-map. , 2015, Lab on a chip.

[16]  Tao Wang,et al.  Exosomal miR-1290 and miR-375 as prognostic markers in castration-resistant prostate cancer. , 2015, European urology.

[17]  Y. Shao,et al.  Plasma long noncoding RNA protected by exosomes as a potential stable biomarker for gastric cancer , 2015, Tumor Biology.

[18]  Xiao-Fan Wang,et al.  A niche role for cancer exosomes in metastasis , 2015, Nature Cell Biology.

[19]  Naoto Tsuchiya,et al.  Circulating Exosomal microRNAs as Biomarkers of Colon Cancer , 2014, PloS one.

[20]  M. Büchler,et al.  Combined evaluation of a panel of protein and miRNA serum‐exosome biomarkers for pancreatic cancer diagnosis increases sensitivity and specificity , 2015, International journal of cancer.

[21]  Muneesh Tewari,et al.  Quantitative and stoichiometric analysis of the microRNA content of exosomes , 2014, Proceedings of the National Academy of Sciences.

[22]  Cesar M. Castro,et al.  Magnetic nanosensor for detection and profiling of erythrocyte-derived microvesicles. , 2013, ACS nano.

[23]  Peter P. Lee,et al.  Tumor-Derived Exosomes Contain microRNAs with Immunological Function: Implications for a Novel Immunosuppression Mechanism. , 2014, MicroRNA.

[24]  D. Chia,et al.  Role of Pancreatic Cancer-derived Exosomes in Salivary Biomarker Development* , 2013, The Journal of Biological Chemistry.

[25]  Simon C Watkins,et al.  Isolation of biologically-active exosomes from human plasma. , 2014, Journal of immunological methods.

[26]  M. Zöller,et al.  Exosomal tumor microRNA modulates premetastatic organ cells. , 2013, Neoplasia.

[27]  J. Lötvall,et al.  Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells , 2007, Nature Cell Biology.

[28]  A. Falus,et al.  Emerging role of extracellular vesicles in inflammatory diseases , 2014, Nature Reviews Rheumatology.

[29]  Jina Ko,et al.  Detection and isolation of circulating exosomes and microvesicles for cancer monitoring and diagnostics using micro-/nano-based devices. , 2016, The Analyst.

[30]  Lynne T. Bemis,et al.  Standardization of sample collection, isolation and analysis methods in extracellular vesicle research , 2013, Journal of extracellular vesicles.

[31]  George A Calin,et al.  Cancer exosomes perform cell-independent microRNA biogenesis and promote tumorigenesis. , 2014, Cancer cell.

[32]  R. Cerione,et al.  Microfluidic isolation of cancer-cell-derived microvesicles from hetergeneous extracellular shed vesicle populations , 2014, Biomedical Microdevices.

[33]  B. Bao,et al.  Exosomes in cancer development, metastasis, and drug resistance: a comprehensive review , 2013, Cancer and Metastasis Reviews.

[34]  A. Guha,et al.  Extracellular vesicles – vehicles that spread cancer genes , 2012, BioEssays : news and reviews in molecular, cellular and developmental biology.

[35]  Robert Langer,et al.  Small RNA combination therapy for lung cancer , 2014, Proceedings of the National Academy of Sciences.

[36]  Sadik Esener,et al.  Recovery of Drug Delivery Nanoparticles from Human Plasma Using an Electrokinetic Platform Technology. , 2015, Small.

[37]  Bernd Giebel,et al.  Exosomes: small vesicles participating in intercellular communication. , 2012, The international journal of biochemistry & cell biology.

[38]  N. Huh,et al.  Noble polymeric surface conjugated with zwitterionic moieties and antibodies for the isolation of exosomes from human serum. , 2012, Bioconjugate chemistry.

[39]  Matt Trau,et al.  Detecting exosomes specifically: a multiplexed device based on alternating current electrohydrodynamic induced nanoshearing. , 2014, Analytical chemistry.

[40]  C. Gilbert,et al.  Discrimination between exosomes and HIV-1: purification of both vesicles from cell-free supernatants. , 2008, Journal of immunological methods.

[41]  He Huang,et al.  Galectin-3 mediates bone marrow microenvironment-induced drug resistance in acute leukemia cells via Wnt/β-catenin signaling pathway , 2015, Journal of Hematology & Oncology.

[42]  Hakho Lee,et al.  Label-free detection and molecular profiling of exosomes with a nano-plasmonic sensor , 2014, Nature Biotechnology.

[43]  K. Pantel,et al.  Increased serum levels of circulating exosomal microRNA-373 in receptor-negative breast cancer patients , 2014, Oncotarget.

[44]  K. Mimori,et al.  Exosomal microRNA in serum is a novel biomarker of recurrence in human colorectal cancer , 2015, British Journal of Cancer.

[45]  D. Atha,et al.  Mechanism of precipitation of proteins by polyethylene glycols. Analysis in terms of excluded volume. , 1981, The Journal of biological chemistry.

[46]  Hakho Lee,et al.  Acoustic purification of extracellular microvesicles. , 2015, ACS nano.

[47]  R. Setterquist,et al.  Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. , 2012, Biochimica et biophysica acta.

[48]  J. Dear,et al.  Quantification of human urinary exosomes by nanoparticle tracking analysis , 2013, The Journal of physiology.

[49]  C. Tacchetti,et al.  Proteome Profiling of Neuroblastoma-Derived Exosomes Reveal the Expression of Proteins Potentially Involved in Tumor Progression , 2013, PloS one.

[50]  H. Nakagawa,et al.  Antibody-coupled monolithic silica microtips for highthroughput molecular profiling of circulating exosomes , 2014, Scientific Reports.

[51]  Romaric Lacroix,et al.  Overcoming limitations of microparticle measurement by flow cytometry. , 2010, Seminars in thrombosis and hemostasis.

[52]  H. Vogel,et al.  Molecular and dimensional profiling of highly purified extracellular vesicles by fluorescence fluctuation spectroscopy. , 2014, Analytical chemistry.

[53]  Jing Liu,et al.  Potential of cancer cell-derived exosomes in clinical application: a review of recent research advances. , 2014, Clinical therapeutics.

[54]  H. Matsubara,et al.  Serum microRNA expression profile: miR-1246 as a novel diagnostic and prognostic biomarker for oesophageal squamous cell carcinoma , 2013, British Journal of Cancer.

[55]  C. Nelson,et al.  Exosomes in Prostate Cancer: Putting Together the Pieces of a Puzzle , 2013, Cancers.

[56]  Matt Trau,et al.  Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing , 2015, Scientific Reports.

[57]  Qun Yu,et al.  Increased Exosomal MicroRNA-21 and MicroRNA-146a Levels in the Cervicovaginal Lavage Specimens of Patients with Cervical Cancer , 2014, International journal of molecular sciences.

[58]  Chao-Min Cheng,et al.  Paper-based immunoaffinity devices for accessible isolation and characterization of extracellular vesicles , 2014 .

[59]  Robert M Hoffman,et al.  Imaging exosome transfer from breast cancer cells to stroma at metastatic sites in orthotopic nude-mouse models. , 2013, Advanced drug delivery reviews.

[60]  Yusuke Yoshioka,et al.  Ultra-sensitive liquid biopsy of circulating extracellular vesicles using ExoScreen , 2014, Nature Communications.

[61]  Elaine C. Campbell,et al.  Nanoparticle tracking analysis monitors microvesicle and exosome secretion from immune cells , 2012, Immunology.

[62]  Willem Stoorvogel,et al.  Fluorescent labeling of nano-sized vesicles released by cells and subsequent quantitative and qualitative analysis by high-resolution flow cytometry , 2012, Nature Protocols.

[63]  Michael J. Heller,et al.  Dielectrophoretic isolation and detection of cancer‐related circulating cell‐free DNA biomarkers from blood and plasma , 2014, Electrophoresis.

[64]  Paolo Bergese,et al.  Colorimetric nanoplasmonic assay to determine purity and titrate extracellular vesicles. , 2015, Analytical chemistry.

[65]  Zuhong Lu,et al.  miRNA in Plasma Exosome is Stable under Different Storage Conditions , 2014, Molecules.

[66]  M. Kester,et al.  Formation and role of exosomes in cancer , 2014, Cellular and Molecular Life Sciences.

[67]  T. Whiteside,et al.  Isolation and Characterization of CD34+ Blast-Derived Exosomes in Acute Myeloid Leukemia , 2014, PloS one.

[68]  Jaesung Park,et al.  Microfluidic filtration system to isolate extracellular vesicles from blood. , 2012, Lab on a chip.

[69]  Graça Raposo,et al.  Extracellular vesicles: Exosomes, microvesicles, and friends , 2013, The Journal of cell biology.

[70]  Valentina R Minciacchi,et al.  Extracellular vesicles in cancer: exosomes, microvesicles and the emerging role of large oncosomes. , 2015, Seminars in cell & developmental biology.

[71]  V. Zhdanov,et al.  Determination of exosome concentration in solution using surface plasmon resonance spectroscopy. , 2014, Analytical chemistry.

[72]  Imre Mäger,et al.  Extracellular vesicles: biology and emerging therapeutic opportunities , 2013, Nature Reviews Drug Discovery.

[73]  M. Gleave,et al.  Reproducibility and efficiency of serum-derived exosome extraction methods. , 2014, Clinical biochemistry.

[74]  Chris Gardiner,et al.  Extracellular vesicle sizing and enumeration by nanoparticle tracking analysis , 2013, Journal of extracellular vesicles.

[75]  Fang Wei,et al.  Detection of exosomal biomarker by electric field-induced release and measurement (EFIRM). , 2013, Biosensors & bioelectronics.

[76]  Michael A. Hollingsworth,et al.  Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver , 2015, Nature Cell Biology.

[77]  T. Oosterkamp,et al.  Determination of the size distribution of blood microparticles directly in plasma using atomic force microscopy and microfluidics , 2012, Biomedical Microdevices.

[78]  Christian Pilarsky,et al.  Glypican-1 identifies cancer exosomes and detects early pancreatic cancer , 2015, Nature.

[79]  Biana Godin,et al.  Ciliated micropillars for the microfluidic-based isolation of nanoscale lipid vesicles. , 2013, Lab on a chip.

[80]  Hongwei Wang,et al.  Expression of Serum Exosomal MicroRNA-21 in Human Hepatocellular Carcinoma , 2014, BioMed research international.

[81]  H. Matsubara,et al.  MicroRNAs in cancer diagnosis and therapy: from bench to bedside , 2013, Surgery Today.

[82]  R. Setterquist,et al.  Methods for the extraction and RNA profiling of exosomes. , 2013, World journal of methodology.

[83]  S. Kristensen,et al.  Extracellular Vesicle (EV) Array: microarray capturing of exosomes and other extracellular vesicles for multiplexed phenotyping , 2013, Journal of extracellular vesicles.

[84]  M. Krumrey,et al.  Innovation in detection of microparticles and exosomes , 2013, Journal of thrombosis and haemostasis : JTH.

[85]  Allan Stensballe,et al.  Diagnostic and prognostic potential of extracellular vesicles in peripheral blood. , 2014, Clinical therapeutics.

[86]  N. Kosaka,et al.  Electrokinetic Evaluation of Individual Exosomes by On-Chip Microcapillary Electrophoresis with Laser Dark-Field Microscopy , 2013 .

[87]  K. Mimori,et al.  Identification of a bona fide microRNA biomarker in serum exosomes that predicts hepatocellular carcinoma recurrence after liver transplantation , 2015, British Journal of Cancer.

[88]  N. Kosaka,et al.  The roles of extracellular vesicles in cancer biology: Toward the development of novel cancer biomarkers , 2014, Proteomics.

[89]  Bernd Giebel,et al.  Characterisation of exosomes derived from human cells by nanoparticle tracking analysis and scanning electron microscopy. , 2011, Colloids and surfaces. B, Biointerfaces.

[90]  Christopher H Contag,et al.  Differential fates of biomolecules delivered to target cells via extracellular vesicles , 2015, Proceedings of the National Academy of Sciences.

[91]  Ger J.A. Arkesteijn,et al.  Quantitative and qualitative flow cytometric analysis of nanosized cell-derived membrane vesicles , 2011, Nanomedicine: Nanotechnology, Biology and Medicine.

[92]  H. Baba,et al.  Clinical impact of serum exosomal microRNA‐21 as a clinical biomarker in human esophageal squamous cell carcinoma , 2013, Cancer.

[93]  Bob S. Carter,et al.  Chip-based analysis of exosomal mRNA mediating drug resistance in glioblastoma , 2015, Nature Communications.

[94]  Qiao Li,et al.  Tumor cell-derived exosomes: a message in a bottle. , 2012, Biochimica et biophysica acta.

[95]  Clotilde Théry,et al.  Biogenesis and secretion of exosomes. , 2014, Current opinion in cell biology.

[96]  Richard J Simpson,et al.  Comparison of ultracentrifugation, density gradient separation, and immunoaffinity capture methods for isolating human colon cancer cell line LIM1863-derived exosomes. , 2012, Methods.