A pharmaceutical investigation into exosomes

Journal of Pharmaceutical Investigation - Tập 48 - Trang 617-626 - 2018
Sumeet Manandhar1, Vinoth Kumar Kothandan2, Jiyun Oh1, Sung Hyun Yoo1, Jinsu Hwang1, Seung Rim Hwang1,2
1College of Pharmacy, Chosun University, Gwangju, Republic of Korea
2Department of Biomedical Sciences, Graduate School, Chosun University, Gwangju, Republic of Korea

Tóm tắt

During the last 5 years, there has been a marked rise in publications regarding exosomes. Exosomes find potential applications as diagnostic biomarkers, therapeutics, drug delivery vehicles, and functional cosmetics and several exosome-based therapies are under clinical trials. Exosomes are nanosized vesicles that contain cell-derived lipid membranes, nucleic acids, and proteins, and are released from the cells of origin, circulate in eukaryotic fluids, and are taken up by recipient cells. The originating cell can be engineered to express specific genes, which enhance immunity, cross-presentation of antigens, or cell targeting by exosomes. Various methods using centrifugation, size-exclusion chromatography, filtration, precipitation, affinity, and microfluidics have been assessed for the isolation and purification of exosomes from biological samples. In addition, exosomes have advantages as drug delivery systems, with biocompatibility, preservation of cargo drugs during circulation in physiological fluids, uptake into the target cells, and drug release after endosomal escape. In this review, we discuss the various aspects of pharmaceutical investigation into exosomes with special focus on engineering, production, characterization, biological activities, applications of exosomes as drug delivery systems, and regulations for evaluation of exosome products.

Tài liệu tham khảo

Alvarez ML, Khosroheidari M, Ravi RK, DiStefano JK (2012) Comparison of protein, microRNA, and mRNA yields using different methods of urinary exosome isolation for the discovery of kidney disease biomarkers. Kidney Int 82:1024–1032 Baranyai T, Herczeg K, Onodi Z, Voszka I, Modos K, Marton N, Nagy G, Mager I, Wood MJ, El Andaloussi S, Palinkas Z, Kumar V, Nagy P, Kittel A, Buzas EI, Ferdinandy P, Giricz Z (2015) Isolation of exosomes from blood plasma: qualitative and quantitative comparison of ultracentrifugation and size exclusion chromatography methods. PLoS One 10:e0145686 Boing AN, van der Pol E, Grootemaat AE, Coumans FA, Sturk A, Nieuwland R (2014) Single-step isolation of extracellular vesicles by size-exclusion chromatography. J Extracell Vesicles 3 Booth AM, Fang Y, Fallon JK, Yang JM, Hildreth JE, Gould SJ (2006) Exosomes and HIV Gag bud from endosome-like domains of the T cell plasma membrane. J Cell Biol 172:923–935 Bosch S, De Beaurepaire L, Allard M, Mosser M, Heichette C, Chrétien D, Jegou D, Bach J-M (2016) Trehalose prevents aggregation of exosomes and cryodamage. Sci Rep Uk 6:36162 Chavez-Munoz C, Morse J, Kilani R, Ghahary A (2008) Primary human keratinocytes externalize stratifin protein via exosomes. J Cell Biochem 104:2165–2173 Cheruvanky A, Zhou H, Pisitkun T, Kopp JB, Knepper MA, Yuen PS, Star RA (2007) Rapid isolation of urinary exosomal biomarkers using a nanomembrane ultrafiltration concentrator. Am J Physiol Renal Physiol 292:F1657-F1661 Choi JS, Yoon HI, Lee KS, Choi YC, Yang SH, Kim IS, Cho YW (2016) Exosomes from differentiating human skeletal muscle cells trigger myogenesis of stem cells and provide biochemical cues for skeletal muscle regeneration. J Control Releas 222:107–115 Clark NA, Lunacek JH, Benedek GB (1970) A study of Brownian motion using light scattering. Am J Phys 38:575–585 Cui Y, Luan J, Li H, Zhou X, Han J (2016) Exosomes derived from mineralizing osteoblasts promote ST2 cell osteogenic differentiation by alteration of microRNA expression. FEBS Lett 590:185–192 de Vrij J, Maas SL, van Nispen M, Sena-Esteves M, Limpens RW, Koster AJ, Leenstra S, Lamfers ML, Broekman ML (2013) Quantification of nanosized extracellular membrane vesicles with scanning ion occlusion sensing. Nanomedicine (Lond) 8:1443–1458 Di Bonito P, Ridolfi B, Columba-Cabezas S, Giovannelli A, Chiozzini C, Manfredi F, Anticoli S, Arenaccio C, Federico M (2015) HPV-E7 delivered by engineered exosomes elicits a protective CD8(+) T cell-mediated immune response. Viruses 7:1079–1099 Di Bonito P, Chiozzini C, Arenaccio C, Anticoli S, Manfredi F, Olivetta E, Ferrantelli F, Falcone E, Ruggieri A, Federico M (2017) Antitumor HPV E7-specific CTL activity elicited by in vivo engineered exosomes produced through DNA inoculation. Int J Nanomed 12:4579–4591 Dukers DF, Meij P, Vervoort MB, Vos W, Scheper RJ, Meijer CJ, Bloemena E, Middeldorp JM (2000) Direct immunosuppressive effects of EBV-encoded latent membrane protein 1. J Immunol 165:663–670 Escola JM, Kleijmeer MJ, Stoorvogel W, Griffith JM, Yoshie O, Geuze HJ (1998) Selective enrichment of tetraspan proteins on the internal vesicles of multivesicular endosomes and on exosomes secreted by human B-lymphocytes. J Biol Chem 273:20121–20127 Filipe V, Hawe A, Jiskoot W (2010) Critical evaluation of nanoparticle tracking analysis (NTA) by NanoSight for the measurement of nanoparticles and protein aggregates. Pharm Res 27:796–810 Geminard C, De Gassart A, Blanc L, Vidal M (2004) Degradation of AP2 during reticulocyte maturation enhances binding of hsc70 and Alix to a common site on TFR for sorting into exosomes. Traffic 5:181–193 Gimona M, Pachler K, Laner-Plamberger S, Schallmoser K, Rohde E (2017) Manufacturing of human extracellular vesicle-based therapeutics for clinical use. Int J Mol Sci 18 Gruenberg J, Van der Goot FG (2006) Mechanisms of pathogen entry through the endosomal compartments. Nat Rev Mol Cell Biol 7:495 Haney MJ, Klyachko NL, Zhao Y, Gupta R, Plotnikova EG, He Z, Patel T, Piroyan A, Sokolsky M, Kabanov AV, Batrakova EV (2015) Exosomes as drug delivery vehicles for Parkinson’s disease therapy. J Control Release 207:18–30 Heijnen HF, Schiel AE, Fijnheer R, Geuze HJ, Sixma JJ (1999) Activated platelets release two types of membrane vesicles: microvesicles by surface shedding and exosomes derived from exocytosis of multivesicular bodies and alpha-granules. Blood 94:3791–3799 Heinemann ML, Ilmer M, Silva LP, Hawke DH, Recio A, Vorontsova MA, Alt E, Vykoukal J (2014) Benchtop isolation and characterization of functional exosomes by sequential filtration. J Chromatogr A 1371:125–135 Illes B, Hirschle P, Barnert S, Cauda V, Wuttke S, Engelke H (2017) Exosome-coated metal-organic framework nanoparticles: an efficient drug delivery platform. Chem Mater 29:8042–8046 Im H, Shao H, Park YI, Peterson VM, Castro CM, Weissleder R, Lee H (2014) Label-free detection and molecular profiling of exosomes with a nano-plasmonic sensor. Nat Biotechnol 32:490–495 Jarad M, Kuczynski E, Morrison J, Viloria-Petit A, Coomber B (2017) Release of endothelial cell associated VEGFR2 during TGF-β modulated angiogenesis in vitro. BMC Cell Biol 18:10 Jeong S, Park J, Pathania D, Castro CM, Weissleder R, Lee H (2016) Integrated magneto-electrochemical sensor for exosome analysis. ACS Nano 10:1802–1809 Jia S, Zocco D, Samuels ML, Chou MF, Chammas R, Skog J, Zarovni N, Momen-Heravi F, Kuo WP (2014) Emerging technologies in extracellular vesicle-based molecular diagnostics. Expert Rev Mol Diagn 14:307–321 Johnstone RM, Adam M, Hammond JR, Orr L, Turbide C (1987) Vesicle formation during reticulocyte maturation. Association of plasma membrane activities with released vesicles (exosomes). J Biol Chem 262:9412–9420 Johnstone RM, Mathew A, Mason AB, Teng K (1991) Exosome formation during maturation of mammalian and avian reticulocytes: evidence that exosome release is a major route for externalization of obsolete membrane proteins. J Cell Physiol 147:27–36 Kanwar SS, Dunlay CJ, Simeone DM, Nagrath S (2014) Microfluidic device (ExoChip) for on-chip isolation, quantification and characterization of circulating exosomes. Lab Chip 14:1891–1900 Keller S, Sanderson MP, Stoeck A, Altevogt P (2006) Exosomes: from biogenesis and secretion to biological function. Immunol Lett 107:102–108 Kesimer M, Gupta R (2015) Physical characterization and profiling of airway epithelial derived exosomes using light scattering. Methods 87:59–63 Kim CH, Lee SG, Kang MJ, Lee S, Choi YW (2017a) Surface modification of lipid-based nanocarriers for cancer cell-specific drug targeting. J Pharm Investig 47:203–227 Kim S, Sohn HJ, Lee HJ, Sohn DH, Hyun SJ, Cho HI, Kim TG (2017b) Use of engineered exosomes expressing HLA and costimulatory molecules to generate antigen-specific CD8 + T cells for adoptive cell therapy. J Immunother 40:83–93 Lai A, Elfeky O, Rice GE, Salomon C (2018) Optimized specific isolation of placenta-derived exosomes from maternal circulation. Methods Mol Biol 1710:131–138 Lakkaraju A, Rodriguez-Boulan E (2008) Itinerant exosomes: emerging roles in cell and tissue polarity. Trends Cell Biol 18:199–209 Lane RE, Korbie D, Anderson W, Vaidyanathan R, Trau M (2015) Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing. Sci Rep 5:7639 Lattanzi L, Federico M (2012) A strategy of antigen incorporation into exosomes: comparing cross-presentation levels of antigens delivered by engineered exosomes and by lentiviral virus-like particles. Vaccine 30:7229–7237 Li XB, Zhang ZR, Schluesener HJ, Xu SQ (2006) Role of exosomes in immune regulation. J Cell Mol Med 10:364–375 Li P, Kaslan M, Lee SH, Yao J, Gao Z (2017) Progress in exosome isolation techniques. Theranostics 7:789–804 Liga A, Vliegenthart AD, Oosthuyzen W, Dear JW, Kersaudy-Kerhoas M (2015) Exosome isolation: a microfluidic road-map. Lab Chip 15:2388–2394 Lim JY, Kim NA, Lim DG, Kim KH, Choi DH, Jeong SH (2016) Process cycle development of freeze drying for therapeutic proteins with stability evaluation. J Pharm Investig 46:519–536 Luan X, Sansanaphongpricha K, Myers I, Chen H, Yuan H, Sun D (2017) Engineering exosomes as refined biological nanoplatforms for drug delivery. Acta Pharmacol Sin 38:754–763 Lyu L, Wang H, Li B, Qin Q, Qi L, Nagarkatti M, Nagarkatti P, Janicki JS, Wang XL, Cui T (2015) A critical role of cardiac fibroblast-derived exosomes in activating renin angiotensin system in cardiomyocytes. J Mol Cell Cardiol 89:268–279 Manfredi F, di Bonito P, Ridolfi B, Anticoli S, Arenaccio C, Chiozzini C, Baz Morelli A, Federico M (2016) The CD8+ T cell-mediated immunity induced by HPV-E6 uploaded in engineered exosomes is improved by ISCOMATRIXTM adjuvant. Vaccines. https://doi.org/10.3390/vaccines4040042 Mignot G, Roux S, Thery C, Ségura E, Zitvogel L (2006) Prospects for exosomes in immunotherapy of cancer. J Cell Mol Med 10:376–388 Moon PG, Lee JE, You S, Kim TK, Cho JH, Kim IS, Kwon TH, Kim CD, Park SH, Hwang D, Kim YL, Baek MC (2011a) Proteomic analysis of urinary exosomes from patients of early IgA nephropathy and thin basement membrane nephropathy. Proteomics 11:2459–2475 Moon PG, You S, Lee JE, Hwang D, Baek MC (2011b) Urinary exosomes and proteomics. Mass Spectrom Rev 30:1185–1202 Morelli AE, Larregina AT, Shufesky WJ, Sullivan ML, Stolz DB, Papworth GD, Zahorchak AF, Logar AJ, Wang Z, Watkins SC, Falo LD Jr, Thomson AW (2004) Endocytosis, intracellular sorting, and processing of exosomes by dendritic cells. Blood 104:3257–3266 Morishita M, Takahashi Y, Matsumoto A, Nishikawa M, Takakura Y (2016) Exosome-based tumor antigens-adjuvant co-delivery utilizing genetically engineered tumor cell-derived exosomes with immunostimulatory CpG DNA. Biomaterials 111:55–65 Munagala R, Aqil F, Jeyabalan J, Gupta RC (2016) Bovine milk-derived exosomes for drug delivery. Cancer Lett 371:48–61 Newton W, Kim J, Luo J, Luo L (2017) Stem cell derived exosomes: a novel vector for tissue repair and diabetic therapy. J Mol Endocrinol. https://doi.org/10.1530/JME-17-0080 Ohno S, Takanashi M, Sudo K, Ueda S, Ishikawa A, Matsuyama N, Fujita K, Mizutani T, Ohgi T, Ochiya T, Gotoh N, Kuroda M (2013) Systemically injected exosomes targeted to EGFR deliver antitumor microRNA to breast cancer cells. Mol Ther 21:185–191 Qazi KR, Torregrosa Paredes P, Dahlberg B, Grunewald J, Eklund A, Gabrielsson S (2010) Proinflammatory exosomes in bronchoalveolar lavage fluid of patients with sarcoidosis. Thorax 65:1016–1024 Qu Y, Franchi L, Nunez G, Dubyak GR (2007) Nonclassical IL-1 beta secretion stimulated by P2 × 7 receptors is dependent on inflammasome activation and correlated with exosome release in murine macrophages. J Immunol 179:1913–1925 Rani S, Ryan AE, Griffin MD, Ritter T (2015) Mesenchymal stem cell-derived extracellular vesicles: toward cell-free therapeutic applications. Mol Ther 23:812–823 Ribeiro MF, Zhu H, Millard RW, Fan G-C (2013) Exosomes function in pro-and anti-angiogenesis. Curr Angiogenes 2:54 Saari H, Lazaro-Ibanez E, Viitala T, Vuorimaa-Laukkanen E, Siljander P, Yliperttula M (2015) Microvesicle- and exosome-mediated drug delivery enhances the cytotoxicity of Paclitaxel in autologous prostate cancer cells. J Control Releas 220:727–737 Sabapatha A, Gercel-Taylor C, Taylor DD (2006) Specific isolation of placenta-derived exosomes from the circulation of pregnant women and their immunoregulatory consequences. Am J Reprod Immunol 56:345–355 Sato YT, Umezaki K, Sawada S, Mukai SA, Sasaki Y, Harada N, Shiku H, Akiyoshi K (2016) Engineering hybrid exosomes by membrane fusion with liposomes. Sci Rep 6:21933 Schorey JS, Bhatnagar S (2008) Exosome function: from tumor immunology to pathogen biology. Traffic 9:871–881 Shao H, Chung J, Lee K, Balaj L, Min C, Carter BS, Hochberg FH, Breakefield XO, Lee H, Weissleder R (2015) Chip-based analysis of exosomal mRNA mediating drug resistance in glioblastoma. Nat Commun 6:6999 Sharma S, Rasool HI, Palanisamy V, Mathisen C, Schmidt M, Wong DT, Gimzewski JK (2010) Structural-mechanical characterization of nanoparticle exosomes in human saliva, using correlative AFM, FESEM, and force spectroscopy. ACS Nano 4:1921–1926 Sheldon H, Heikamp E, Turley H, Dragovic R, Thomas P, Oon CE, Leek R, Edelmann M, Kessler B, Sainson RC (2010) New mechanism for Notch signaling to endothelium at a distance by Delta-like 4 incorporation into exosomes. Blood 116:2385–2394 Sheridan C (2016) Exosome cancer diagnostic reaches market. Nat Biotechnol 34:359–360 Shin S, Han D, Park MC, Mun JY, Choi J, Chun H, Kim S, Hong JW (2017) Separation of extracellular nanovesicles and apoptotic bodies from cancer cell culture broth using tunable microfluidic systems. Sci Rep 7:9907 Shtam TA, Kovalev RA, Varfolomeeva EY, Makarov EM, Kil YV, Filatov MV (2013) Exosomes are natural carriers of exogenous siRNA to human cells in vitro. Cell Commun Signal 11:88 Simeoli R, Montague K, Jones HR, Castaldi L, Chambers D, Kelleher JH, Vacca V, Pitcher T, Grist J, Al-Ahdal H, Wong LF, Perretti M, Lai J, Mouritzen P, Heppenstall P, Malcangio M (2017) Exosomal cargo including microRNA regulates sensory neuron to macrophage communication after nerve trauma. Nat Commun 8:1778 Smith JA, Ng KS, Mead BE, Dopson S, Reeve B, Edwards J, Wood MJA, Carr AJ, Bure K, Karp JM, Brindley DA (2015) Extracellular vesicles: commercial potential as byproducts of cell manufacturing for research and therapeutic use. Bioprocess Int. http://www.bioprocessintl.com/manufacturing/cell-therapies/extracellularvesicles-commercial-potential-as-byproducts-of-cell-manufacturing-for-research-and-therapeutic-use. Accessed 14 April 2015 Song JG, Lee SH, Han H-K (2017) The stabilization of biopharmaceuticals: current understanding and future perspectives. J Pharm Investig 47:475–496 Sun D, Zhuang X, Xiang X, Liu Y, Zhang S, Liu C, Barnes S, Grizzle W, Miller D, Zhang HG (2010) A novel nanoparticle drug delivery system: the anti-inflammatory activity of curcumin is enhanced when encapsulated in exosomes. Mol Ther 18:1606–1614 Tatischeff I, Larquet E, Falcon-Perez JM, Turpin PY, Kruglik SG (2012) Fast characterisation of cell-derived extracellular vesicles by nanoparticles tracking analysis, cryo-electron microscopy, and Raman tweezers microspectroscopy. J Extracell Vesicles 1 Tauro BJ, Greening DW, Mathias RA, Ji H, Mathivanan S, Scott AM, Simpson RJ (2012) Comparison of ultracentrifugation, density gradient separation, and immunoaffinity capture methods for isolating human colon cancer cell line LIM1863-derived exosomes. Methods 56:293–304 Thery C, Regnault A, Garin J, Wolfers J, Zitvogel L, Ricciardi-Castagnoli P, Raposo G, Amigorena S (1999) Molecular characterization of dendritic cell-derived exosomes. Selective accumulation of the heat shock protein hsc73. J Cell Biol 147:599–610 Thery C, Amigorena S, Raposo G, Clayton A (2006) Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol Chap 3:Unit 3 22 Tian Y, Li S, Song J, Ji T, Zhu M, Anderson GJ, Wei J, Nie G (2014) A doxorubicin delivery platform using engineered natural membrane vesicle exosomes for targeted tumor therapy. Biomaterials 35:2383–2390 Tibbitt MW, Dahlman JE, Langer R (2016) Emerging frontiers in drug delivery. J Am Chem Soc 138:704–717 van Niel G, Raposo G, Candalh C, Boussac M, Hershberg R, Cerf-Bensussan N, Heyman M (2001) Intestinal epithelial cells secrete exosome-like vesicles. Gastroenterology 121:337–349 van Niel G, Porto-Carreiro I, Simoes S, Raposo G (2006) Exosomes: a common pathway for a specialized function. J Biochem 140:13–21 van der Pol E, Hoekstra AG, Sturk A, Otto C, van Leeuwen TG, Nieuwland R (2010) Optical and non-optical methods for detection and characterization of microparticles and exosomes. J Thromb Haemost 8:2596–2607 van der Pol E, Boing AN, Harrison P, Sturk A, Nieuwland R (2012) Classification, functions, and clinical relevance of extracellular vesicles. Pharmacol Rev 64:676–705 Vlassov AV, Magdaleno S, Setterquist R, Conrad R (2012) Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. Biochimica et Biophysica Acta (BBA) Gen Subj 1820:940–948 Wahlgren J, De LKT, Brisslert M, Vaziri Sani F, Telemo E, Sunnerhagen P, Valadi H (2012) Plasma exosomes can deliver exogenous short interfering RNA to monocytes and lymphocytes. Nucl Acids Res 40:e130 Wan Y, Cheng G, Liu X, Hao SJ, Nisic M, Zhu CD, Xia YQ, Li WQ, Wang ZG, Zhang WL, Rice SJ, Sebastian A, Albert I, Belani CP, Zheng SY (2017) Rapid magnetic isolation of extracellular vesicles via lipid-based nanoprobes. Nat Biomed Eng. https://doi.org/10.1038/s41551-017-0058 Wang Y, Zhang L, Li Y, Chen L, Wang X, Guo W, Zhang X, Qin G, He SH, Zimmerman A, Liu Y, Kim IM, Weintraub NL, Tang Y (2015) Exosomes/microvesicles from induced pluripotent stem cells deliver cardioprotective miRNAs and prevent cardiomyocyte apoptosis in the ischemic myocardium. Int J Cardiol 192:61–69 Whitford W, Ludlow JW, Cadwell JJ (2015) Continuous production of exosomes: utilizing the technical advantages of hollow-fiber bioreactor technology. Genet Eng Biotechnol News 35:34 Wiklander OP, Nordin JZ, O’Loughlin A, Gustafsson Y, Corso G, Mager I, Vader P, Lee Y, Sork H, Seow Y, Heldring N, Alvarez-Erviti L, Smith CI, Le Blanc K, Macchiarini P, Jungebluth P, Wood MJ, Andaloussi SE (2015) Extracellular vesicle in vivo biodistribution is determined by cell source, route of administration and targeting. J Extracell Vesicles 4:26316 Witwer KW, Buzas EI, Bemis LT, Bora A, Lässer C, Lötvall J, Nolte-′t Hoen EN, Piper MG, Sivaraman S, Skog J, Théry C, Wauben MH, Hochberg F (2013) Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. J Extracell Vesicles. https://doi.org/10.3402/jev.v2i0.20360 Wu M, Ouyang Y, Wang Z, Zhang R, Huang PH, Chen C, Li H, Li P, Quinn D, Dao M, Suresh S, Sadovsky Y, Huang TJ (2017) Isolation of exosomes from whole blood by integrating acoustics and microfluidics. Proc Natl Acad Sci USA 114:10584–10589 Xie Y, Bai O, Zhang H, Li W, Xiang J (2010a) Tumor necrosis factor gene-engineered J558 tumor cell-released exosomes stimulate tumor antigen P1A-specific CD8 + CTL responses and antitumor immunity. Cancer Biother Radiopharm 25:21–28 Xie Y, Bai O, Zhang H, Yuan J, Zong S, Chibbar R, Slattery K, Qureshi M, Wei Y, Deng Y, Xiang J (2010b) Membrane-bound HSP70-engineered myeloma cell-derived exosomes stimulate more efficient CD8(+) CTL- and NK-mediated antitumour immunity than exosomes released from heat-shocked tumour cells expressing cytoplasmic HSP70. J Cell Mol Med 14:2655–2666 Xu R, Greening DW, Zhu HJ, Takahashi N, Simpson RJ (2016) Extracellular vesicle isolation and characterization: toward clinical application. J Clin Invest 126:1152–1162 Xu J-Y, Chen G-H, Yang Y-J (2017) Exosomes: a rising star in failing hearts. Front Physiol 8:494 Yang T, Martin P, Fogarty B, Brown A, Schurman K, Phipps R, Yin VP, Lockman P, Bai S (2015) Exosome delivered anticancer drugs across the blood-brain barrier for brain cancer therapy in Danio rerio. Pharm Res 32:2003–2014 Yang Y, Hong Y, Nam GH, Chung JH, Koh E, Kim IS (2017) Virus-mimetic fusogenic exosomes for direct delivery of integral membrane proteins to target cell membranes. Adv Mater 29 Yim N, Ryu SW, Choi K, Lee KR, Lee S, Choi H, Kim J, Shaker MR, Sun W, Park JH, Kim D, Heo WD, Choi C (2016) Exosome engineering for efficient intracellular delivery of soluble proteins using optically reversible protein-protein interaction module. Nat Commun 7:12277 Zarovni N, Corrado A, Guazzi P, Zocco D, Lari E, Radano G, Muhhina J, Fondelli C, Gavrilova J, Chiesi A (2015) Integrated isolation and quantitative analysis of exosome shuttled proteins and nucleic acids using immunocapture approaches. Methods 87:46–58 Zeringer E, Barta T, Li M, Vlassov AV (2015) Strategies for isolation of exosomes. Cold Spring Harb Protoc 2015: 319–323 Zitvogel L, Regnault A, Lozier A, Wolfers J, Flament C, Tenza D, Ricciardi-Castagnoli P, Raposo G, Amigorena S (1998) Eradication of established murine tumors using a novel cell-free vaccine: dendritic cell derived exosomes. Nat Med 4:594–600 Zou G, Benktander JD, Gizaw ST, Gaunitz S, Novotny MV (2017) Comprehensive analytical approach toward glycomic characterization and profiling in urinary exosomes. Anal Chem 89:5364–5372