Abstract
Extracellular vesicles (EVs) are considered to be a new generation of bioinspired nanoscale drug delivery systems due to their low immunogenicity, natural functionality, and excellent biocompatibility. However, limitations such as low uptake efficiency, insufficient production, and inhomogeneous performance undermine their potential. To address these issues, numerous researchers have put forward various methods and applications for enhancing EV uptake in recent decades. In this review, we introduce various methods for the cellular uptake of EVs and summarize recent advances on the methods and mechanisms for enhancing EV uptake. In addition, we provide further understanding regarding enhancing EV uptake and put forward prospects and challenges for the development of EV-based therapy in the future.
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Pegtel DM, Gould SJ. Exosomes. Annu Rev Biochem, 2019,88(1):487–514
El Andaloussi S, Mäger I, Breakefield XO, et al. Extracellular vesicles: biology and emerging therapeutic opportunities. Nat Rev Drug Discov, 2013,12(5):347–357
Ettelaie C, Collier MEW, Maraveyas A, et al. Characterization of physical properties of tissue factor-containing microvesicles and a comparison of ultracentrifuge-based recovery procedures. J Extracell Vesicles, 2014,3(1):1
Battistelli M, Falcieri E. Apoptotic Bodies: Particular Extracellular Vesicles Involved in Intercellular Communication. Biology, 2020,9(1):21
Elmore S. Apoptosis: A Review of Programmed Cell Death. Toxicol Pathol, 2007,35(4):495–516
Mathieu M, Martin-Jaular L, Lavieu G, et al. Specificities of secretion and uptake of exosomes and other extracellular vesicles for cell-to-cell communication. Nat Cell Biol, 2019,21(1):9–17
Colombo M, Raposo G, Thery C. Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Annu Rev Cell Dev Biol, 2014,30:255–289
Maas SLN, Breakefield XO, Weaver AM. Extracellular Vesicles: Unique Intercellular Delivery Vehicles. Trends Cell Biol, 2017,27(3):172–188
Abels ER, Breakefield XO. Introduction to Extracellular Vesicles: Biogenesis, RNA Cargo Selection, Content, Release, and Uptake. Cell Mol Neurobiol, 2016,36(3):301–312
Nolte-’T Hoen ENM, Buschow SI, Anderton SM, et al. Activated T cells recruit exosomes secreted by dendritic cells via LFA-1. Blood, 2009,113(9):1977–1981
Zhang B, Wang M, Gong A, et al. HucMSC-Exosome Mediated-Wnt4 Signaling Is Required for Cutaneous Wound Healing. Stem Cells, 2015,33(7):2158–2168
Cui X, He Z, Liang Z, et al. Exosomes From Adipose-derived Mesenchymal Stem Cells Protect the Myocardium Against Ischemia/Reperfusion Injury Through Wnt/β-Catenin Signaling Pathway. J Cardiovasc Pharmacol, 2017,70(4):225–231
Ratajczak J, Miekus K, Kucia M, et al. Embryonic stem cell-derived microvesicles reprogram hematopoietic progenitors: evidence for horizontal transfer of mRNA and protein delivery. Leukemia, 2006,20(5):847–856
Men Y, Yelick J, Jin S, et al. Exosome reporter mice reveal the involvement of exosomes in mediating neuron to astroglia communication in the CNS. Nat Commun, 2019,10(1):4136
Bang C, Batkai S, Dangwal S, et al. Cardiac fibroblast-derived microRNA passenger strand-enriched exosomes mediate cardiomyocyte hypertrophy. J Clin Invest, 2014,124(5):2136–2146
Zamani P, Fereydouni N, Butler AE, et al. The therapeutic and diagnostic role of exosomes in cardiovascular diseases. Trends Cardiovasc Med, 2019,29(6):313–323
Howitt J, Hill AF. Exosomes in the Pathology of Neurodegenerative Diseases. J Biol Chem, 2016,291(52):26589–26597
Chen N, Sun XY, Ding ZC, et al. Small Extracellular Vesicles Secreted by Peri-urethral Tissues Regulate Fibroblast Function and Contribute to the Pathogenesis of Female Stress Urinary Incontinence. Curr Med Sci, 2023,43(4):803–810
Duan CY, Fan WL, Chen F. Roles of Optineurin and Extracellular Vesicles in Glaucomatous Retinal Cell Loss. Curr Med Sci, 2023,43(2):367–375
Osaki M, Okada F. Exosomes and Their Role in Cancer Progression. Yonago Acta Med, 2019,62(2):182–190
Deng Z, Liu Y, Liu C, et al. Immature myeloid cells induced by a high-fat diet contribute to liver inflammation. Hepatology, 2009,50(5):1412–1420
Nakase I, Kobayashi NB, Takatani-Nakase T, et al. Active macropinocytosis induction by stimulation of epidermal growth factor receptor and oncogenic Ras expression potentiates cellular uptake efficacy of exosomes. Sci Rep, 2015,5(1):10300
Nakase I, Futaki S. Combined treatment with a pH-sensitive fusogenic peptide and cationic lipids achieves enhanced cytosolic delivery of exosomes. Sci Rep, 2015,5(1):10112
Xu H, Liao C, Liang S, et al. A Novel Peptide-Equipped Exosomes Platform for Delivery of Antisense Oligonucleotides. ACS Appl Mater Interfaces, 2021,13(9):10760–10767
Martínez-Santillán A, González-Valdez J. Novel Technologies for Exosome and Exosome-like Nanovesicle Procurement and Enhancement. Biomedicines, 2023,11(5):1487
Tran NH, Nguyen DD, Nguyen NM, et al. Dualtargeting exosomes for improved drug delivery in breast cancer. Nanomedrcme (Lond), 2023,18(7):599–611
Luan X, Sansanaphongpricha K, Myers I, et al. Engineering exosomes as refined biological nanoplatforms for drug delivery. Acta Pharmacol Sin, 2017,38(6):754–763
Tian T, Wang Y, Wang H, et al. Visualizing of the cellular uptake and intracellular trafficking of exosomes by live-cell microscopy. J Cell Biochem, 2010,111(2):488–496
Tian T, Zhu YL, Hu FH, et al. Dynamics of exosome internalization and trafficking. J Cell Physiol, 2013,228(7):1487–1495
Sokolova V, Ludwig A, Hornung S, et al. Characterisation of exosomes derived from human cells by nanoparticle tracking analysis and scanning electron microscopy. Colloids and Surfaces B: Biointerfaces, 2011,87(1):146–150
Takahashi Y, Nishikawa M, Shinotsuka H, et al. Visualization and in vivo tracking of the exosomes of murine melanoma B16-BL6 cells in mice after intravenous injection. J Biotechnol, 2013,165(2):77–84
van Niel G, D’Angelo G, Raposo G. Shedding light on the cell biology of extracellular vesicles. Nat Rev Mol Cell Biol, 2018,19(4):213–228
Gyorgy B, Hung ME, Breakefield XO, et al. Therapeutic applications of extracellular vesicles: clinical promise and open questions. Annu Rev Pharmacol Toxicol, 2015,55:439–464
Gurung S, Perocheau D, Touramanidou L, et al. The exosome journey: from biogenesis to uptake and intracellular signalling. Cell Commun Signal, 2021,19(1):47
Parolini I, Federici C, Raggi C, et al. Microenvironmental pH Is a Key Factor for Exosome Traffic in Tumor Cells. J Biol Chem, 2009,284(49):34211–34222
Montecalvo A, Larregina AT, Shufesky WJ, et al. Mechanism of transfer of functional microRNAs between mouse dendritic cells via exosomes. Blood, 2012,119(3):756–766
Morelli AE, Larregina AT, Shufesky WJ, et al. Endocytosis, intracellular sorting, and processing of exosomes by dendritic cells. Blood, 2004,104(10):3257–3266
Escrevente C, Keller S, Altevogt P, et al. Interaction and uptake of exosomes by ovarian cancer cells. BMC Cancer, 2011,11(1):108
Araldi RP, Delvalle DA, Da Costa VR, et al. Exosomes as a Nano-Carrier for Chemotherapeutics: A New Era of Oncology. Cells, 2023,12(17):2144
Gonda A, Kabagwira J, Senthil GN, et al. Internalization of Exosomes through Receptor-Mediated Endocytosis. Mol Cancer Res, 2019,17(2):337–347
Mckelvey KJ, Powell KL, Ashton AW, et al. Exosomes: Mechanisms of Uptake. J Circ Biomark, 2015,4:7
Mettlen M, Chen P, Srinivasan S, et al. Regulation of Clathrin-Mediated Endocytosis. Annu Rev Biochem, 2018,87(1):871–896
Kiss AL, Botos E. Endocytosis via caveolae: alternative pathway with distinct cellular compartments to avoid lysosomal degradation? J Cell Mol Med, 2009,13(7):1228–1237
Barres C, Blanc L, Bette-Bobillo P, et al. Galectin-5 is bound onto the surface of rat reticulocyte exosomes and modulates vesicle uptake by macrophages. Blood, 2010,115(3):696–705
Nanbo A, Kawanishi E, Yoshida R, et al. Exosomes derived from Epstein-Barr virus-infected cells are internalized via caveola-dependent endocytosis and promote phenotypic modulation in target cells. J Virol, 2013,87(18):10334–10347
Menck K, Klemm F, Gross JC, et al. Induction and transport of Wnt 5a during macrophage-induced malignant invasion is mediated by two types of extracellular vesicles. Oncotarget, 2013,4(11):2057–2066
Izquierdo-Useros N, Naranjo-Gómez M, Archer J, et al. Capture and transfer of HIV-1 particles by mature dendritic cells converges with the exosome-dissemination pathway. Blood, 2009,113(12):2732–2741
Nabi IR, Le PU. Caveolae/raft-dependent endocytosis. The Journal of Cell Biology, 2003,161(4):673–677
Doherty GJ, Mcmahon HT. Mechanisms of endocytosis. Annu Rev Biochem, 2009,78:857–902
Feng D, Zhao W, Ye Y, et al. Cellular Internalization of Exosomes Occurs Through Phagocytosis. Traffic, 2010,11(5):675–687
Gordon S. Phagocytosis: An Immunobiologic Process. Immunity, 2016,44(3):463–475
Lim JP, Gleeson PA. Macropinocytosis: an endocytic pathway for internalising large gulps. Immunol Cell Biol, 2011,89(8):836–843
Heusermann W, Hean J, Trojer D, et al. Exosomes surf on filopodia to enter cells at endocytic hot spots, traffic within endosomes, and are targeted to the ER. J Cell Biol, 2016,213(2):173–184
Mattila PK, Lappalainen P. Filopodia: molecular architecture and cellular functions. Nat Rev Mol Cell Biol, 2008,9(6):446–454
Lehmann MJ, Sherer NM, Marks CB, et al. Actin- and myosin-driven movement of viruses along filopodia precedes their entry into cells. J Cell Biol, 2005,170(2):317–325
Barrès C, Blanc L, Bette-Bobillo P, et al. Galectin-5 is bound onto the surface of rat reticulocyte exosomes and modulates vesicle uptake by macrophages. Blood, 2010,115(3):696–705
Fitzner D, Schnaars M, van Rossum D, et al. Selective transfer of exosomes from oligodendrocytes to microglia by macropinocytosis. J Cell Sci, 2011,124(3):447–458
Frühbeis C, Fröhlich D, Kuo WP, et al. Neurotransmitter-Triggered Transfer of Exosomes Mediates Oligod-endrocyte-Neuron Communication. PLoS Biol, 2013,11(7):e1001604
Nanbo A, Kawanishi E, Yoshida R, et al. Exosomes Derived from Epstein-Barr Virus-Infected Cells Are Internalized via Caveola-Dependent Endocytosis and Promote Phenotypic Modulation in Target Cells. J Virol, 2013,87(18):10334–10347
Svensson KJ, Christianson HC, Wittrup A, et al. Exosome Uptake Depends on ERK1/2-Heat Shock Protein 27 Signaling and Lipid Raft-mediated Endocytosis Negatively Regulated by Caveolin-1. J Biol Chem, 2013,288(24):17713–17724
Rejman J, Oberle V, Zuhorn IS, et al. Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis. Biochem J, 2004,377 (Pt 1):159–169
Mcmahon HT, Boucrot E. Molecular mechanism and physiological functions of clathrin-mediated endocytosis. Nat Rev Mol Cell Biol, 2011,12(8):517–533
Wang Z, Tiruppathi C, Cho J, et al. Delivery of nanoparticle: complexed drugs across the vascular endothelial barrier via caveolae. IUBMB Life, 2011,63(8):659–667
Akinc A, Battaglia G. Exploiting endocytosis for nanomedicines. Cold Spring Harb Perspect Biol, 2013,5(11):a16980
Costa Verdera H, Gitz-Francois JJ, Schiffelers RM, et al. Cellular uptake of extracellular vesicles is mediated by clathrin-independent endocytosis and macropinocytosis. J Control Release, 2017,266:100–108
Ye H, Wang F, Xu G, et al. Advancements in engineered exosomes for wound repair: current research and future perspectives. Front Bioeng Biotechnol, 2023,11:1301362
Nakagawa Y, Arafiles JVV, Kawaguchi Y, et al. Stearylated Macropinocytosis-Inducing Peptides Facilitating the Cellular Uptake of Small Extracellular Vesicles. Bioconjug Chem, 2022,33(5):869–880
Nakase I, Noguchi K, Fujii I, et al. Vectorization of biomacromolecules into cells using extracellular vesicles with enhanced internalization induced by macropinocytosis. Sci Rep, 2016,6(1):34937
Hirase S, Aoki A, Hattori Y, et al. Dodecaborate-Encapsulated Extracellular Vesicles with Modification of Cell-Penetrating Peptides for Enhancing Macropinocytotic Cellular Uptake and Biological Activity in Boron Neutron Capture Therapy. Mol Pharm, 2022,19(4):1135–1145
Noguchi K, Obuki M, Sumi H, et al. Macropinocytosis-Inducible Extracellular Vesicles Modified with Antimicrobial Protein CAP18-Derived Cell-Penetrating Peptides for Efficient Intracellular Delivery. Mol Pharm, 2021,18(9):3290–3301
Shimoda A, Miura R, Tateno H, et al. Assessment of Surface Glycan Diversity on Extracellular Vesicles by Lectin Microarray and Glycoengineering Strategies for Drug Delivery Applications. Small Methods, 2022,6(2):e2100785
Nishida-Aoki N, Tominaga N, Kosaka N, et al. Altered biodistribution of deglycosylated extracellular vesicles through enhanced cellular uptake. J Extracell Vesicles, 2020,9(1):1713527
Zhan Q, Yi K, Li X, et al. Phosphatidylcholine-Engineered Exosomes for Enhanced Tumor Cell Uptake and Intracellular Antitumor Drug Delivery. Macromol Biosci, 2021,21(8):e2100042
Zou J, Shi M, Liu X, et al. Aptamer-Functionalized Exosomes: Elucidating the Cellular Uptake Mechanism and the Potential for Cancer-Targeted Chemotherapy. Anal Chem, 2019,91(3):2425–2430
Hade MD, Suire CN, Suo Z. An Effective Peptide-Based Platform for Efficient Exosomal Loading and Cellular Delivery of a microRNA. ACS Appl Mater Interfaces, 2023,15(3):3851–3866
Liao Z, Liu H, Ma L, et al. Engineering Extracellular Vesicles Restore the Impaired Cellular Uptake and Attenuate Intervertebral Disc Degeneration. ACS Nano, 2021,15(9):14709–14724
Meyer C, Losacco J, Stickney Z, et al. Pseudotyping exosomes for enhanced protein delivery in mammalian cells. Int J Nanomedicine, 2017,12:3153–3170
Temchura VV, Tenbusch M, Nchinda G, et al. Enhancement of immunostimulatory properties of exosomal vaccines by incorporation of fusion-competent G protein of vesicular stomatitis virus. Vaccine, 2008,26(29):3662–3672
Hung ME, Leonard JN. Stabilization of Exosometargeting Peptides via Engineered Glycosylation. J Biol Chem, 2015,290(13):8166–8172
Zhang J, Song H, Dong Y, et al. Surface Engineering of HEK293 Cell-Derived Extracellular Vesicles for Improved Pharmacokinetic Profile and Targeted Delivery of IL-12 for the Treatment of Hepatocellular Carcinoma. Int J Nanomedicine, 2023,18:209–223
Kelemen A, Carmi I, Oszvald Á, et al. IFITM1 expression determines extracellular vesicle uptake in colorectal cancer. Cell Mol Life Sci, 2021,78(21–22):7009–7024
Zhang S, Guo M, Guo T, et al. DAL-1/4.1B promotes the uptake of exosomes in lung cancer cells via Heparan Sulfate Proteoglycan 2 (HSPG2). Mol Cell Biochem, 2022,477(1):241–254
Hazawa M, Tomiyama K, Saotome-Nakamura A, et al. Radiation increases the cellular uptake of exosomes through CD29/CD81 complex formation. Biochem Biophys Res Commun, 2014,446(4):1165–1171
Mizuta R, Sasaki Y, Kawasaki R, et al. Magnetically Navigated Intracellular Delivery of Extracellular Vesicles Using Amphiphilic Nanogels. Bioconjug Chem, 2019,30(8):2150–2155
Gong C, Zhang X, Shi M, et al. Tumor Exosomes Reprogrammed by Low pH Are Efficient Targeting Vehicles for Smart Drug Delivery and Personalized Therapy against their Homologous Tumor. Adv Sci (Weinh), 2021,8(10):2002787
Kim H, Kang J, Mun D, et al. Calcium chloride enhances the delivery of exosomes. PLoS One, 2019,14(7):e220036
Ferrero-Andrés A, Closa D, Roselló-Catafau J, et al. Polyethylene Glycol 35 (PEG35) Modulates Exosomal Uptake and Function. Polymers (Basel), 2020,12(12):3044
Ma S, Song L, Bai Y, et al. Improved intracellular delivery of exosomes by surface modification with fluorinated peptide dendrimers for promoting angiogenesis and migration of HUVECs. RSC Adv, 2023,13(17):11269–11277
Matsuki Y, Yanagawa T, Sumiyoshi H, et al. Modification of exosomes with carbonate apatite and a glycan polymer improves transduction efficiency and target cell selectivity. Biochem Biophys Res Commun, 2021,583:93–99
Takenaka T, Nakai S, Katayama M, et al. Effects of gefitinib treatment on cellular uptake of extracellular vesicles in EGFR-mutant non-small cell lung cancer cells. Int J Pharm, 2019,572:118762
Zhao Z, Mcgill J, Gamero-Kubota P, et al. Microfluidic on-demand engineering of exosomes towards cancer immunotherapy. Lab Chip, 2019,19(10):1877–1886
Bui S, Dancourt J, Lavieu G. Virus-Free Method to Control and Enhance Extracellular Vesicle Cargo Loading and Delivery. ACS Appl Bio Mater, 2023,6(3):1081–1091
Wu P, Tang Y, Jin C, et al. Neutrophil membrane engineered HucMSC sEVs alleviate cisplatin-induced AKI by enhancing cellular uptake and targeting. J Nanobiotechnology, 2022,20(1):353
Hu S, Wang X, Li Z, et al. Platelet membrane and stem cell exosome hybrids enhance cellular uptake and targeting to heart injury. Nano Today, 2021,39:101210
Shao J, Zaro J, Shen Y. Advances in Exosome-Based Drug Delivery and Tumor Targeting: From Tissue Distribution to Intracellular Fate. Int J Nanomedicine, 2020,15:9355–9371
Gonda A, Kabagwira J, Senthil GN, et al. Exosomal survivin facilitates vesicle internalization. Oncotarget, 2018,9(79):34919–34934
Tkach M, Kowal J, Zucchetti AE, et al. Qualitative differences in T-cell activation by dendritic cell-derived extracellular vesicle subtypes. The EMBO Journal, 2017,36(20):3012–3028
Guan S, Li Q, Liu P, et al. Experimental immunology Umbilical cord blood-derived dendritic cells loaded with BGC823 tumor antigens and DC-derived exosomes stimulate efficient cytotoxic T-lymphocyte responses and antitumor immunity in vitro and in vivo. Cent Eur J Immunol, 2014,2(2):142–151
Sobo-Vujanovic A, Munich S, Vujanovic NL. Dendritic-cell exosomes cross-present Toll-like receptor-ligands and activate bystander dendritic cells. Cell Immunol, 2014,289(1–2):119–127
Munich S, Sobo-Vujanovic A, Buchser WJ, et al. Dendritic cell exosomes directly kill tumor cells and activate natural killer cells via TNF superfamily ligands. Oncoimmunology, 2014,1(7):1074–1083
Zheng Y, Tu C, Zhang J, et al. Inhibition of multiple myeloma-derived exosomes uptake suppresses the functional response in bone marrow stromal cell. Int J Oncol, 2019,54(3):1061–1070
Buschow SI, Nolte T Hoen ENM, Van Niel G, et al. MHC II in Dendritic Cells is Targeted to Lysosomes or T Cell-Induced Exosomes Via Distinct Multivesicular Body Pathways. Traffic, 2009,10(10):1528–1542
Verweij FJ, Bebelman MP, Jimenez CR, et al. Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling. J Cell Biol, 2018,217(3):1129–1142
Horibe S, Tanahashi T, Kawauchi S, et al. Mechanism of recipient cell-dependent differences in exosome uptake. BMC Cancer, 2018,18(1):47
Eguchi S, Takefuji M, Sakaguchi T, et al. Cardiomyocytes capture stem cell-derived, anti-apoptotic microRNA-214 via clathrin-mediated endocytosis in acute myocardial infarction. J Biol Chem, 2019,294(31):11665–11674
Benmerah A, Bayrou M, Cerf-Bensussan N, et al. Inhibition of clathrin-coated pit assembly by an Eps15 mutant. J Cell Sci, 1999,112(9):1303–1311
Yoon JH, Ashktorab H, Smoot DT, et al. Uptake and tumor-suppressive pathways of exosome-associated GKN1 protein in gastric epithelial cells. Gastric Cancer, 2020,23(5):848–862
Koumangoye RB, Sakwe AM, Goodwin JS, et al. Detachment of Breast Tumor Cells Induces Rapid Secretion of Exosomes Which Subsequently Mediate Cellular Adhesion and Spreading. PLoS One, 2011,6(9):e24234
Skotland T, Hessvik NP, Sandvig K, et al. Exosomal lipid composition and the role of ether lipids and phosphoinositides in exosome biology. J Lipid Res, 2019,60(1):9–18
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The authors declare that there is no conflict of interest with any financial organization or corporation or individual that can inappropriately influence this work.
Author Zhen-bing CHEN is a member of the Editorial Board for Current Medical Science. The paper was handled by the other editors and has undergone rigorous peer review process. Author Zhen-bing CHEN was not involved in the journal’s review of, or decisions related to, this manuscript.
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This work was supported by the National Natural Science Foundation of China (No. 82370838 and No. 82172221).
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Xu, Yp., Jiang, T., Yang, Xf. et al. Methods, Mechanisms, and Application Prospects for Enhancing Extracellular Vesicle Uptake. CURR MED SCI 44, 247–260 (2024). https://doi.org/10.1007/s11596-024-2861-7
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DOI: https://doi.org/10.1007/s11596-024-2861-7