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Cited by 537Extracellular vesicles in diseaseBarrier Structure and Function StudiesNanoparticle-Based Drug Delivery

Elucidation of Exosome Migration Across the Blood–Brain Barrier Model In Vitro

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Chen, Claire C., Liu, Linan, Ma, Fengxia, Wong, Chi W., Guo, Xuning E., Chacko, Jenu V., Farhoodi, Henry P., Zhang, Shirley X. +7 more·2016·Cellular and Molecular Bioengineering
DOI 10.1007/s12195-016-0458-3PMC PMC5382965
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Abstract

The delivery of therapeutics to the central nervous system (CNS) remains a major challenge in part due to the presence of the blood-brain barrier (BBB). Recently, cell-derived vesicles, particularly exosomes, have emerged as an attractive vehicle for targeting drugs to the brain, but whether or how they cross the BBB remains unclear. Here, we investigated the interactions between exosomes and brain microvascular endothelial cells (BMECs) in vitro under conditions that mimic the healthy and inflamed BBB in vivo. Transwell assays revealed that luciferase-carrying exosomes can cross a BMEC monolayer under stroke-like, inflamed conditions (TNF-α activated) but not under normal conditions. Confocal microscopy showed that exosomes are internalized by BMECs through endocytosis, co-localize with endosomes, in effect primarily utilizing the transcellular route of crossing. Together, these results indicate that cell-derived exosomes can cross the BBB model under stroke-like conditions in vitro. This study encourages further development of engineered exosomes as drug delivery vehicles or tracking tools for treating or monitoring neurological diseases.

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Crossref · 10.1007/s12195-016-0458-3

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  • Exosomes utilize two primary routes to traverse the blood-brain barrier: paracellular and transcellular.

    “EXO were found to utilize two primary routes to traverse the BBB: paracellular, which involves tight and adherens junctions, and transcellular, which involves micropinocytosis, receptor, or caveola-mediated endocytosis [30].”

    From Oral Microbially-Induced Small Extracellular Vesicles Cross the Blood–Brain Barrier (2024) · cites this paper as [30]

    In Results and Discussion (page unavailable), the authors report that paracellular diffusion contributed little to exosome crossing of their in vitro BBB model, while transcellular transport accounted for most migration. The paper therefore does not support describing paracellular and transcellular transport as two primary routes used by exosomes. (LLM, conf=0.88)

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  • The engineered exosomes could cross the multiple layers of the blood-brain-barrier and target cancer cells within the brain tissue.

    “Importantly, the engineered exosomes could cross the multiple layers of the blood-brain-barrier (BBB) and target cancer cells within the brain tissue (40, 65–68).”

    From Potential Therapeutic Effects of Exosomes Packed With a miR-21-Sponge Construct in a Rat Model of Glioblastoma (2019) · cites this paper as [65]

    The Results and Discussion report that engineered exosomes crossed a single brain microvascular endothelial cell monolayer in vitro under TNF-α-induced, stroke-like conditions. The Conclusions say that astrocytes, pericytes, and basement membrane were not included in that model. The paper does not show passage through the BBB’s multiple layers or targeting of cancer cells in brain tissue. (LLM, conf=0.90)

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  1. [2]Alvarez-Erviti L Seow Y Yin H Betts C Lakhal S Wood MJ Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes Nat Biotechnol 2011 29 341 345 21423189
  2. [3]Andreone BJ Lacoste B Gu C Neuronal and vascular interactions Annu Rev Neurosci 2015 38 25 46 25782970 PMC5729758
  3. [4]Banks WA From blood-brain barrier to blood-brain interface: new opportunities for CNS drug delivery Nat Rev Drug Discov 2016 26794270
  4. [5]Chen J Zhang ZG Li Y Wang L Xu YX Gautam SC Lu M Zhu Z Chopp M Intravenous administration of human bone marrow stromal cells induces angiogenesis in the ischemic boundary zone after stroke in rats Circ Res 2003 92 692 699 12609969
  5. [6]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 25288114
  6. [7]Deli MA Descamps L Dehouck MP Cecchelli R Joo F Abraham CS Torpier G Exposure of tumor necrosis factor-alpha to luminal membrane of bovine brain capillary endothelial cells cocultured with astrocytes induces a delayed increase of permeability and cytoplasmic stress fiber formation of actin J Neurosci Res 1995 41 717 726 7500373
  7. [8]Dendrou CA Fugger L Friese MA Immunopathology of multiple sclerosis Nat Rev Immunol 2015 15 545 558 26250739
  8. [9]Descamps L Cecchelli R Torpier G Effects of tumor necrosis factor on receptor-mediated endocytosis and barrier functions of bovine brain capillary endothelial cell monolayers J Neuroimmunol 1997 74 173 184 9119971
  9. [11]Dulamea AO The potential use of mesenchymal stem cells in stroke therapy-From bench to bedside Journal of the Neurological Sciences 2015 352 1 11 25818674
  10. [12]El-Andaloussi S Mager I Breakefield XO Wood MJ Extracellular vesicles: biology and emerging therapeutic opportunities Nat Rev Drug Discov 2013 12 347 357 23584393
  11. [13]Etame AB Diaz RJ Smith CA Mainprize TG Hynynen K Rutka JT Focused ultrasound disruption of the blood-brain barrier: a new frontier for therapeutic delivery in molecular neurooncology Neurosurg Focus 2012 32 E3 PMC4106119 22208896
  12. [14]Faille D El-Assaad F Mitchell AJ Alessi MC Chimini G Fusai T Grau GE Combes V Endocytosis and intracellular processing of platelet microparticles by brain endothelial cells J Cell Mol Med 2012 16 1731 1738 21883894 PMC3822686
  13. [15]Fischer S Wiesnet M Marti HH Renz D Schaper W Simultaneous activation of several second messengers in hypoxia-induced hyperpermeability of brain derived endothelial cells J Cell Physiol 2004 198 359 369 14755541
  14. [17]Gabathuler R Approaches to transport therapeutic drugs across the blood-brain barrier to treat brain diseases Neurobiol Dis 2010 37 48 57 19664710
  15. [18]Ge QY Zhou YX Lu JF Bai YF Xie XY Lu ZH miRNA in Plasma Exosome is Stable under Different Storage Conditions Molecules 2014 19 1568 1575 24473213 PMC6271968
  16. [19]Hsu J Rappaport J Muro S Specific binding, uptake, and transport of ICAM-1-targeted nanocarriers across endothelial and subendothelial cell components of the blood-brain barrier Pharm Res 2014 31 1855 1866 24558007 PMC4065215
  17. [20]Imai T Takahashi Y Nishikawa M Kato K Morishita M Yamashita T Matsumoto A Charoenviriyakul C Takakura Y Macrophage-dependent clearance of systemically administered B16BL6-derived exosomes from the blood circulation in mice J Extracell Vesicles 2015 4 26238 25669322 PMC4323410
  18. [21]Kalani A Tyagi A Tyagi N Exosomes: mediators of neurodegeneration, neuroprotection and therapeutics Mol Neurobiol 2014 49 590 600 23999871 PMC3951279
  19. [22]Komarova Y Malik AB Regulation of endothelial permeability via paracellular and transcellular transport pathways Annu Rev Physiol 2010 72 463 493 20148685
  20. [23]Kourembanas S Exosomes: vehicles of intercellular signaling, biomarkers, and vectors of cell therapy Annu. Rev. Physiol 2015 77 13 27 25293529
  21. [24]Lai CP Mardini O Ericsson M Prabhakar S Maguire CA Chen JW Tannous BA Breakefield XO Dynamic Biodistribution of Extracellular Vesicles in Vivo Using a Multimodal Imaging Reporter Acs Nano 2014 8 483 494 24383518 PMC3934350
  22. [25]Lasser C Exosomes in diagnostic and therapeutic applications: biomarker, vaccine and RNA interference delivery vehicle Expert Opin. Biol. Ther 2015 15 103 117 25363342
  23. [26]Lasser C O’Neil SE Ekerljung L Ekstrom K Sjostrand M Lotvall J RNA-containing exosomes in human nasal secretions Am J Rhinol Allergy 2011 25 89 93 21172122
  24. [27]Lee CC MacKay JA Frechet JM Szoka FC Designing dendrimers for biological applications Nat Biotechnol 2005 23 1517 1526 16333296
  25. [28]Lee RH Pulin AA Seo MJ Kota DJ Ylostalo J Larson BL Semprun-Prieto L Delafontaine P Prockop DJ Intravenous hMSCs improve myocardial infarction in mice because cells embolized in lung are activated to secrete the anti-inflammatory protein TSG-6 Cell Stem Cell 2009 5 54 63 19570514 PMC4154377
  26. [29]Liao W Pham V Liu L Riazifar M Pone EJ Zhang SX Ma F Lu M Walsh CM Zhao W Mesenchymal stem cells engineered to express selectin ligands and IL-10 exert enhanced therapeutic efficacy in murine experimental autoimmune encephalomyelitis Biomaterials 2016 77 87 97 26584349 PMC4684451
  27. [30]Liu L Eckert MA Riazifar H Kang DK Agalliu D Zhao W From blood to the brain: can systemically transplanted mesenchymal stem cells cross the blood-brain barrier? Stem Cells Int 2013 2013 435093 23997771 PMC3753739
  28. [31]Liu L Zhang SX Aeran R Liao W Lu M Polovin G Pone EJ Zhao W Exogenous marker-engineered mesenchymal stem cells detect cancer and metastases in a simple blood assay Stem Cell Res Ther 2015 6 181 26391980 PMC4578609
  29. [32]Manders EMM Verbeek FJ Aten JA Measurement of Colocalization of Objects in Dual-Color Confocal Images Journal of Microscopy-Oxford 1993 169 375 382 33930978
  30. [33]Mathivanan S Ji H Simpson RJ Exosomes: extracellular organelles important in intercellular communication J Proteomics 2010 73 1907 1920 20601276
  31. [34]Mulcahy LA Pink RC Carter DR Routes and mechanisms of extracellular vesicle uptake J Extracell Vesicles 2014 3 PMC4122821 25143819
  32. [36]Ohno S Takanashi M Sudo K Ueda S Ishikawa A Matsuyama N Fujita K Mizutani T Ohgi T Ochiya T Gotoh N Kuroda M Systemically injected exosomes targeted to EGFR deliver antitumor microRNA to breast cancer cells Mol Ther 2013 21 185 191 PMC3538304 23032975
  33. [37]Ostrowski M Carmo NB Krumeich S Fanget I Raposo G Savina A Moita CF Schauer K Hume AN Freitas RP Goud B Benaroch P Hacohen N Fukuda M Desnos C Seabra MC Darchen F Amigorena S Moita LF Thery C Rab27a and Rab27b control different steps of the exosome secretion pathway Nat Cell Biol 2010 12 supp 1– 13 19 30 19966785
  34. [38]Pardridge WM Targeted delivery of protein and gene medicines through the blood-brain barrier Clin Pharmacol Ther 2015 97 347 361 25669455
  35. [39]Pardridge WM Blood-brain barrier endogenous transporters as therapeutic targets: a new model for small molecule CNS drug discovery Expert Opin Ther Targets 2015 19 1059 1072 25936389
  36. [40]Pardridge WM Drug transport across the blood-brain barrier J Cereb Blood Flow Metab 2012 32 1959 1972 22929442 PMC3494002
  37. [41]Pardridge WM The blood-brain barrier: bottleneck in brain drug development NeuroRx 2005 2 3 14 15717053 PMC539316
  38. [42]Peer D Karp JM Hong S Farokhzad OC Margalit R Langer R Nanocarriers as an emerging platform for cancer therapy Nat Nanotechnol 2007 2 751 760 18654426
  39. [43]Petros RA DeSimone JM Strategies in the design of nanoparticles for therapeutic applications Nat. Rev. Drug. Discov 2010 9 615 627 20616808
  40. [44]Pols MS Klumperman J Trafficking and function of the tetraspanin CD63 Exp Cell Res 2009 315 1584 1592 18930046
  41. [45]Raposo G Stoorvogel W Extracellular vesicles: Exosomes, microvesicles, and friends Journal of Cell Biology 2013 200 373 383 23420871 PMC3575529
  42. [46]Ridder K Keller S Dams M Rupp AK Schlaudraff J Del Turco D Starmann J Macas J Karpova D Devraj K Depboylu C Landfried B Arnold B Plate KH Hoglinger G Sultmann H Altevogt P Momma S Extracellular vesicle-mediated transfer of genetic information between the hematopoietic system and the brain in response to inflammation Plos Biology 2014 12 PMC4043485 24893313
  43. [47]Rochfort KD Collins LE Murphy RP Cummins PM Downregulation of blood-brain barrier phenotype by proinflammatory cytokines involves NADPH oxidase-dependent ROS generation: consequences for interendothelial adherens and tight junctions PLoS One 2014 9 e101815 24992685 PMC4081725
  44. [48]Rubin LL Staddon JM The cell biology of the blood-brain barrier Annu Rev Neurosci 1999 22 11 28 10202530
  45. [49]Sandoval KE Witt KA Blood-brain barrier tight junction permeability and ischemic stroke Neurobiol Dis 2008 32 200 219 18790057
  46. [50]Schiera G Di Liegro CM Di Liegro I Extracellular membrane vesicles as vehicles for brain cell-to-cell interactions in physiological as well as pathological conditions Biomed Research International 2015 PMC4637152 26583089
  47. [51]Schnitzer JE Oh P Pinney E Allard J Filipin-sensitive caveolae-mediated transport in endothelium: reduced transcytosis, scavenger endocytosis, and capillary permeability of select macromolecules J Cell Biol 1994 127 1217 1232 7525606 PMC2120262
  48. [52]Shlosberg D Benifla M Kaufer D Friedman A Blood-brain barrier breakdown as a therapeutic target in traumatic brain injury Nature Reviews Neurology 2010 6 393 403 20551947 PMC3625732
  49. [53]Song J Ischemic apoplexy-induced sequelae treated by penetrating puncture with long needles J Tradit Chin Med 2002 22 200 202 12400428
  50. [54]Sordi V Malosio ML Marchesi F Mercalli A Melzi R Giordano T Belmonte N Ferrari G Leone BE Bertuzzi F Zerbini G Allavena P Bonifacio E Piemonti L Bone marrow mesenchymal stem cells express a restricted set of functionally active chemokine receptors capable of promoting migration to pancreatic islets Blood 2005 106 419 427 15784733
  51. [55]Svensson KJ Christianson HC Wittrup A Bourseau-Guilmain E Lindqvist E Svensson LM Morgelin M Belting M 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 17713 17724 23653359 PMC3682571
  52. [56]Takahashi Y Nishikawa M Shinotsuka H Matsui Y Ohara S Imai T Takakura Y Visualization and in vivo tracking of the exosomes of murine melanoma B16-BL6 cells in mice after intravenous injection J Biotechnol 2013 165 77 84 23562828
  53. [57]Takeshita Y Obermeier B Cotleur A Sano Y Kanda T Ransohoff RM An in vitro blood-brain barrier model combining shear stress and endothelial cell/astrocyte co-culture J Neurosci Methods 2014 232 165 172 24858797 PMC4122330
  54. [58]Tannous BA Kim DE Fernandez JL Weissleder R Breakefield XO Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo Mol Ther 2005 11 435 443 15727940
  55. [60]Thery C Ostrowski M Segura E Membrane vesicles as conveyors of immune responses Nat Rev Immunol 2009 9 581 593 19498381
  56. [61]Thery C Zitvogel L Amigorena S Exosomes: composition, biogenesis and function Nat Rev Immunol 2002 2 569 579 12154376
  57. [63]Tian T Zhu YL Zhou YY Liang GF Wang YY Hu FH Xiao ZD Exosome uptake through clathrin-mediated endocytosis and macropinocytosis and mediating miR-21 delivery J Biol Chem 2014 289 22258 22267 24951588 PMC4139237
  58. [64]Torchilin VP Multifunctional, stimuli-sensitive nanoparticulate systems for drug delivery Nat. Rev. Drug Discov 2014 13 813 827 25287120 PMC4489143
  59. [66]Trumpp A Essers M Wilson A Awakening dormant haematopoietic stem cells Nat Rev Immunol 2010 10 201 209 20182459
  60. [67]Upadhyay RK Drug delivery systems, CNS protection, and the blood brain barrier Biomed Res Int 2014 2014 869269 25136634 PMC4127280
  61. [68]Valadi H Ekstrom K Bossios A Sjostrand M Lee JJ Lotvall JO Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells Nat Cell Biol 2007 9 654 659 17486113
  62. [69]Vercauteren D Vandenbroucke RE Jones AT Rejman J Demeester J De Smedt SC Sanders NN Braeckmans K The use of inhibitors to study endocytic pathways of gene carriers: optimization and pitfalls Mol Ther 2010 18 561 569 20010917 PMC2839427
  63. [70]Wajant H Pfizenmaier K Scheurich P Tumor necrosis factor signaling Cell Death Differ 2003 10 45 65 12655295
  64. [71]Whalen GF Shing Y Folkman J The fate of intravenously administered bFGF and the effect of heparin Growth Factors 1989 1 157 164 2624780
  65. [72]Wolburg H Lippoldt A Tight junctions of the blood-brain barrier: development, composition and regulation Vascul Pharmacol 2002 38 323 337 12529927
  66. [73]Xin H Li Y Chopp M Exosomes/miRNAs as mediating cell-based therapy of stroke Front Cell Neurosci 2014 8 377 25426026 PMC4226157
  67. [74]Xin H Li Y Cui Y Yang JJ Zhang ZG Chopp M Systemic administration of exosomes released from mesenchymal stromal cells promote functional recovery and neurovascular plasticity after stroke in rats J Cereb Blood Flow Metab 2013 33 1711 1715 23963371 PMC3824189
  68. [75]Yang T Martin P Fogarty B Brown A Schurman K Phipps R Yin VP Lockman P Bai S Exosome delivered anticancer drugs across the blood-brain barrier for brain cancer therapy in Danio rerio Pharm Res 2015 32 2003 2014 25609010 PMC4520542
  69. [76]Zhang Y Chopp M Meng Y Katakowski M Xin H Mahmood A Xiong Y Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury J Neurosurg 2015 122 856 867 25594326 PMC4382456
  70. [77]Zhuang X Xiang X Grizzle W Sun D Zhang S Axtell RC Ju S Mu J Zhang L Steinman L Miller D Zhang HG Treatment of brain inflammatory diseases by delivering exosome encapsulated anti-inflammatory drugs from the nasal region to the brain Mol Ther 2011 19 1769 1779 21915101 PMC3188748

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Authors
Chen, C. C., Liu, L., Ma, F., Wong, C. W., Guo, X. E., Chacko, J. V., Farhoodi, H. P., Zhang, S. X., Zimak, J., Ségaliny, A., Riazifar, M., Pham, V., Digman, M. A., Pone, E. J., & Zhao, W.
Affiliations
  • University of California, Irvine (US)
  • Chinese Academy of Medical Sciences & Peking Union Medical College (CN)
Corresponding author
Zhao, Weian
ORCID
Venue
Cellular and Molecular Bioengineering
Publisher
Springer Science and Business Media LLC
Published
2016-07-07
Volume
9
Issue
4
Pages
509-529
ISSN
  • 1865-5025 (ISSN-L)
  • 1865-5033
DOI
10.1007/s12195-016-0458-3
PMC
PMC5382965
OpenAlex
W2465939084
Cited by
537
References
7
Citations by year
2026: 54 · 2025: 59 · 2024: 66 · 2023: 71 · 2022: 78 · 2021: 83 · 2020: 60 · 2019: 35 · 2018: 23 · 2017: 8
FWCI
11.44
Type
journal article
Language
en
Open access
Licence
Locations
  • Cellular and Molecular Bioengineering (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion, cc-by)
  • eScholarship (California Digital Library) (submittedVersion, cc-by)
  • RUNE (Research UNE) (submittedVersion)
Funders
  • National Institutes of Health (1DP2CA195763-01, NHLBI T32)
  • National Science Foundation (GRFP)
  • American Heart Association (13BGIA17140099)
  • UC Irvine Academic Senate Council on Research, Computing, and Libraries (SIIG-2013-2014-25)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
Keywords
exosomes, blood-brain barrier, brain microvascular endothelial cells, endocytosis, TNF-α-induced inflammation, engineered exosomes, brain drug delivery

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