Advertisement

The AAPS Journal

, 20:67 | Cite as

The Phenotypic Effects of Exosomes Secreted from Distinct Cellular Sources: a Comparative Study Based on miRNA Composition

  • Scott Ferguson
  • Sera Kim
  • Christine Lee
  • Michael Deci
  • Juliane Nguyen
Research Article Theme: Pioneering Pharmaceutical Science by Emerging Investigators
Part of the following topical collections:
  1. Theme: Pioneering Pharmaceutical Science by Emerging Investigators

Abstract

Exosomes are nano-sized vesicles composed of lipids, proteins, and nucleic acids. Their molecular landscape is diverse, and exosomes derived from different cell types have distinct biological activities. Since exosomes are now being utilized as delivery vehicles for exogenous therapeutic cargoes, their intrinsic properties and biological effects must be understood. We performed miRNA profiling and found substantial differences in the miRNA landscape of prostate cancer (PC3) and human embryonic kidney (HEK) 293 exosomes with little correlation in abundance of common miRNAs (R2 = 0.16). Using a systems-level bioinformatics approach, the most abundant miRNAs in PC3 exosomes but not HEK exosomes were predicted to significantly modulate integrin signaling, with integrin-β3 loss inducing macrophage M2 polarization. PC3 but not HEK exosomes downregulated integrin-β3 expression levels by 70%. There was a dose-dependent polarization of RAW 264.7 macrophages toward an M2 phenotype when treated with PC3-derived exosomes but not HEK-derived exosomes. Conversely, HEK exosomes, widely utilized as delivery vehicles, were predicted to target cadherin signaling, with experimental validation showing a significant increase in the migratory potential of MCF7 breast cancer cells treated with HEK exosomes. Even widely utilized exosomes are unlikely to be inert, and their intrinsic activity ought to be assessed before therapeutic deployment.

KEY WORDS

exosomes miRNA profiling phenotypic effects M2 polarization cadherin signaling 

Notes

Acknowledgements

We acknowledge support by the NIH through awards R01EB023262 and R21EB021454. We thank Dr. Prashant Singh for performing the miRNA profiling for HEK and PC3 cells and exosomes. The authors declare no competing financial interests.

Author’s Contribution

J.N. and S.F. conceived and designed the experiments. S.F. carried out the exosomal characterization, and cellular uptake experiment. S.K. carried out the bioinformatics analysis and MTS assay. C.J.L. performed the macrophage polarization study. M.D. provided M2 polarization insights and reviewed the manuscript. J.N. and S.F. analyzed the data. S.F., and J.N. wrote, reviewed and edited the manuscript. J.N supervised the project. All authors approved the final manuscript.

Supplementary material

12248_2018_227_MOESM1_ESM.docx (98 kb)
ESM 1 (DOCX 97 kb)

References

  1. 1.
    Ferguson SW, Nguyen J. Exosomes as therapeutics: the implications of molecular composition and exosomal heterogeneity. J Control Release. 2016;228:179–90.CrossRefPubMedGoogle Scholar
  2. 2.
    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(6):654–U72.CrossRefPubMedGoogle Scholar
  3. 3.
    Lu J, Clark AG. Impact of microRNA regulation on variation in human gene expression. Genome Res. 2012;22(7):1243–54.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Quesenberry PJ, Aliotta J, Deregibus MC, Camussi G. Role of extracellular RNA-carrying vesicles in cell differentiation and reprogramming. Stem Cell Res Ther. 2015;6:153.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Gallo A, Tandon M, Alevizos I, Illei GG. The majority of microRNAs detectable in serum and saliva is concentrated in exosomes. Plos One. 2012;7(3).Google Scholar
  6. 6.
    Mulcahy LA, Pink RC, Carter DR. Routes and mechanisms of extracellular vesicle uptake. J Extracell Vesicles 2014;3.Google Scholar
  7. 7.
    Franzen CA, Simms PE, Van Huis AF, Foreman KE, Kuo PC, Gupta GN. Characterization of uptake and internalization of exosomes by bladder cancer cells. Biomed Res Int. 2014;2014:619829.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    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(4):341–5.CrossRefPubMedGoogle Scholar
  9. 9.
    Chen L, Charrier A, Zhou Y, Chen R, Yu B, Agarwal K, et al. Epigenetic regulation of connective tissue growth factor by microRNA-214 delivery in exosomes from mouse or human hepatic stellate cells. Hepatology. 2014;59(3):1118–29.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Cooper JM, Wiklander PB, Nordin JZ, Al-Shawi R, Wood MJ, Vithlani M, et al. Systemic exosomal siRNA delivery reduced alpha-synuclein aggregates in brains of transgenic mice. Mov Disord. 2014;29(12):1476–85.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Katakowski M, Buller B, Zheng X, Lu Y, Rogers T, Osobamiro O, et al. Exosomes from marrow stromal cells expressing miR-146b inhibit glioma growth. Cancer Lett. 2013;335(1):201–4.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Liu Y, Li D, Liu Z, Zhou Y, Chu D, Li X, et al. Targeted exosome-mediated delivery of opioid receptor Mu siRNA for the treatment of morphine relapse. Sci Rep. 2015;5:17543.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Munoz JL, Bliss SA, Greco SJ, Ramkissoon SH, Ligon KL, Rameshwar P. Delivery of functional anti-miR-9 by mesenchymal stem cell-derived exosomes to glioblastoma multiforme cells conferred chemosensitivity. Mol Ther Nucleic Acids. 2013;2:e126.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Ohno S, Takanashi M, Sudo K, Ueda S, Ishikawa A, Matsuyama N, et al. Systemically injected exosomes targeted to EGFR deliver antitumor microRNA to breast cancer cells. Mol Ther. 2013;21(1):185–91.CrossRefPubMedGoogle Scholar
  15. 15.
    Kim MS, Haney MJ, Zhao Y, Mahajan V, Deygen I, Klyachko NL, et al. Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells. Nanomedicine. 2016;12(3):655–64.CrossRefPubMedGoogle Scholar
  16. 16.
    Batrakova EV, Kim MS. Development and regulation of exosome-based therapy products. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2016;8(5):744–57.CrossRefPubMedGoogle Scholar
  17. 17.
    Harris DA, Patel SH, Gucek M, Hendrix A, Westbroek W, Taraska JW. Exosomes released from breast cancer carcinomas stimulate cell movement. PLoS One. 2015;10(3):e0117495.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Liu Y, Gu Y, Han Y, Zhang Q, Jiang Z, Zhang X, et al. Tumor exosomal RNAs promote lung pre-metastatic niche formation by activating alveolar epithelial TLR3 to recruit neutrophils. Cancer Cell. 2016;30(2):243–56.CrossRefPubMedGoogle Scholar
  19. 19.
    Ye SB, Li ZL, Luo DH, Huang BJ, Chen YS, Zhang XS, et al. Tumor-derived exosomes promote tumor progression and T-cell dysfunction through the regulation of enriched exosomal microRNAs in human nasopharyngeal carcinoma. Oncotarget. 2014;5(14):5439–52.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Zhuang G, Wu X, Jiang Z, Kasman I, Yao J, Guan Y, et al. Tumour-secreted miR-9 promotes endothelial cell migration and angiogenesis by activating the JAK-STAT pathway. EMBO J. 2012;31(17):3513–23.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Tadokoro H, Umezu T, Ohyashiki K, Hirano T, Ohyashiki JH. Exosomes derived from hypoxic leukemia cells enhance tube formation in endothelial cells. J Biol Chem. 2013;288(48):34343–51.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Fong MY, Zhou W, Liu L, Alontaga AY, Chandra M, Ashby J, et al. Breast-cancer-secreted miR-122 reprograms glucose metabolism in premetastatic niche to promote metastasis. Nat Cell Biol. 2015;17(2):183–94.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Zhou W, Fong MY, Min Y, Somlo G, Liu L, Palomares MR, et al. Cancer-secreted miR-105 destroys vascular endothelial barriers to promote metastasis. Cancer Cell. 2014;25(4):501–15.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Azmi AS, Bao B, Sarkar FH. Exosomes in cancer development, metastasis, and drug resistance: a comprehensive review. Cancer Metastasis Rev. 2013;32(3–4):623–42.CrossRefPubMedGoogle Scholar
  25. 25.
    Zhang X, Yuan X, Shi H, Wu L, Qian H, Xu W. Exosomes in cancer: small particle, big player. J Hematol Oncol. 2015;8:83.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Camussi G, Cantaluppi V, Deregibus MC, Gatti E, Tetta C. Role of microvesicles in acute kidney injury. Contrib Nephrol. 2011;174:191–9.CrossRefPubMedGoogle Scholar
  27. 27.
    Barile L, Lionetti V, Cervio E, Matteucci M, Gherghiceanu M, Popescu LM, et al. Extracellular vesicles from human cardiac progenitor cells inhibit cardiomyocyte apoptosis and improve cardiac function after myocardial infarction. Cardiovasc Res. 2014;103(4):530–41.CrossRefPubMedGoogle Scholar
  28. 28.
    Lai RC, Arslan F, Lee MM, Sze NS, Choo A, Chen TS, et al. Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury. Stem Cell Res. 2010;4(3):214–22.CrossRefPubMedGoogle Scholar
  29. 29.
    Zhang Z, Yang J, Yan W, Li Y, Shen Z, Asahara T. Pretreatment of cardiac stem cells with exosomes derived from mesenchymal stem cells enhances myocardial repair. J Am Heart Assoc. 2016;5(1).Google Scholar
  30. 30.
    Zhao Y, Sun X, Cao W, Ma J, Sun L, Qian H, et al. Exosomes derived from human umbilical cord mesenchymal stem cells relieve acute myocardial ischemic injury. Stem Cells Int. 2015;2015:761643.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Katsuda T, Ochiya T. Molecular signatures of mesenchymal stem cell-derived extracellular vesicle-mediated tissue repair. Stem Cell Res Ther. 2015;6:212.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Ferguson SW, Wang J, Lee CJ, Liu M, Neelamegham S, Canty JM, et al. The microRNA regulatory landscape of MSC-derived exosomes: a systems view. Sci Rep. 2018;8(1):1419.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Graham FL, Smiley J, Russell WC, Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977;36(1):59–74.CrossRefPubMedGoogle Scholar
  34. 34.
    Feng D, Zhao WL, Ye YY, Bai XC, Liu RQ, Chang LF, et al. Cellular internalization of exosomes occurs through phagocytosis. Traffic. 2010;11(5):675–87.CrossRefPubMedGoogle Scholar
  35. 35.
    Tominaga N, Hagiwara K, Kosaka N, Honma K, Nakagama H, Ochiya T. RPN2-mediated glycosylation of tetraspanin CD63 regulates breast cancer cell malignancy. Mol Cancer. 2014;13.Google Scholar
  36. 36.
    Kowal J, Arras G, Colombo M, Jouve M, Morath JP, Primdal-Bengtson B, et al. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proc Natl Acad Sci. 2016;113(8):E968–E77.CrossRefPubMedGoogle Scholar
  37. 37.
    Sokolova V, Ludwig AK, Hornung S, Rotan O, Horn PA, Epple M, et al. Characterisation of exosomes derived from human cells by nanoparticle tracking analysis and scanning electron microscopy. Colloids Surf B Biointerfaces. 2011;87(1):146–50.CrossRefPubMedGoogle Scholar
  38. 38.
    Ohshima H. Encyclopedia of biocolloid and biointerface science. Hoboken, New Jersey: John Wiley & Sons, Inc.; 2016. volumes cm p.Google Scholar
  39. 39.
    Jablonski KA, Amici SA, Webb LM, Ruiz-Rosado Jde D, Popovich PG, Partida-Sanchez S, et al. Novel markers to delineate murine M1 and M2 macrophages. PLoS One. 2015;10(12):e0145342.CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Li C, Levin M, Kaplan DL. Bioelectric modulation of macrophage polarization. Sci Rep. 2016;6:21044.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Banerjee S, Xie N, Cui H, Tan Z, Yang S, Icyuz M, et al. MicroRNA let-7c regulates macrophage polarization. J Immunol. 2013;190(12):6542–9.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Hazan RB, Phillips GR, Qiao RF, Norton L, Aaronson SA. Exogenous expression of N-cadherin in breast cancer cells induces cell migration, invasion, and metastasis. J Cell Biol. 2000;148(4):779–90.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Cai J, Guan H, Fang L, Yang Y, Zhu X, Yuan J, et al. MicroRNA-374a activates Wnt/beta-catenin signaling to promote breast cancer metastasis. J Clin Invest. 2013;123(2):566–79.PubMedPubMedCentralGoogle Scholar
  44. 44.
    Cha BH, Shin SR, Leijten J, Li YC, Singh S, Liu JC, et al. Integrin-mediated interactions control macrophage polarization in 3D hydrogels. Adv Healthc Mater. 2017;6(21).  https://doi.org/10.1002/adhm.201700289.
  45. 45.
    Su XM, Esser AK, Amend SR, Xiang JY, Xu YL, Ross MH, et al. Antagonizing integrin beta 3 increases immunosuppression in cancer. Cancer Res. 2016;76(12):3484–95.CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Zhang LP, Dong YJ, Dong YL, Cheng JZ, Du J. Role of integrin-beta 3 protein in macrophage polarization and regeneration of injured muscle. J Biol Chem. 2012;287(9):6177–86.CrossRefPubMedGoogle Scholar
  47. 47.
    Muller DW, Bosserhoff AK. Integrin beta(3) expression is regulated by let-7a miRNA in malignant melanoma. Oncogene. 2008;27(52):6698–706.CrossRefPubMedGoogle Scholar
  48. 48.
    Rath M, Muller I, Kropf P, Closs EI, Munder M. Metabolism via arginase or nitric oxide synthase: two competing arginine pathways in macrophages. Front Immunol. 2014;5:532.CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Mantovani A, Sozzani S, Locati M, Allavena P, Sica A. Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes. Trends Immunol. 2002;23(11):549–55.CrossRefPubMedGoogle Scholar
  50. 50.
    Soki FN, Koh AJ, Jones JD, Kim YW, Dai JL, Keller ET, et al. Polarization of prostate cancer-associated macrophages is induced by milk fat globule-EGF factor 8 (MFG-E8)-mediated efferocytosis. J Biol Chem. 2014;289(35):24560–72.CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Chen PC, Cheng HC, Wang J, Wang SW, Tai HC, Lin CW, et al. Prostate cancer-derived CCN3 induces M2 macrophage infiltration and contributes to angiogenesis in prostate cancer microenvironment. Oncotarget. 2014;5(6):1595–608.CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Su MJ, Aldawsari H, Amiji M. Pancreatic cancer cell exosome-mediated macrophage reprogramming and the role of microRNAs 155 and 125b2 transfection using nanoparticle delivery systems. Sci Rep 2016;6:30110.  https://doi.org/10.1038/srep30110.
  53. 53.
    Saha B, Momen-Heravi F, Kodys K, Szabo G. MicroRNA cargo of extracellular vesicles from alcohol-exposed monocytes signals naive monocytes to differentiate into M2 macrophages with increased phagocytic activity. Eur J Clin Investig. 2016;46:82–3.Google Scholar
  54. 54.
    Liu CY, Lin HH, Tang MJ, Wang YK. Vimentin contributes to epithelial-mesenchymal transition cancer cell mechanics by mediating cytoskeletal organization and focal adhesion maturation. Oncotarget. 2015;6(18):15966–83.PubMedPubMedCentralGoogle Scholar
  55. 55.
    Nieman MT, Prudoff RS, Johnson KR, Wheelock MJ. N-cadherin promotes motility in human breast cancer cells regardless of their E-cadherin expression. J Cell Biol. 1999;147(3):631–44.CrossRefPubMedPubMedCentralGoogle Scholar
  56. 56.
    Hiraguri S, Godfrey T, Nakamura H, Graff J, Collins C, Shayesteh L, et al. Mechanisms of inactivation of E-cadherin in breast cancer cell lines. Cancer Res. 1998;58(9):1972–7.PubMedGoogle Scholar
  57. 57.
    Taherian A, Li X, Liu Y, Haas TA. Differences in integrin expression and signaling within human breast cancer cells. BMC Cancer. 2011;11:293.CrossRefPubMedPubMedCentralGoogle Scholar
  58. 58.
    Hurst NJ Jr, Najy AJ, Ustach CV, Movilla L, Kim HR. Platelet-derived growth factor-C (PDGF-C) activation by serine proteases: implications for breast cancer progression. Biochem J. 2012;441(3):909–18.CrossRefPubMedPubMedCentralGoogle Scholar
  59. 59.
    Wang C, Bian Z, Wei D, Zhang JG. miR-29b regulates migration of human breast cancer cells. Mol Cell Biochem. 2011;352(1–2):197–207.CrossRefPubMedGoogle Scholar
  60. 60.
    Tang YT, Huang YY, Zheng L, Qin SH, Xu XP, An TX, et al. Comparison of isolation methods of exosomes and exosomal RNA from cell culture medium and serum. Int J Mol Med. 2017;40(3):834–44.CrossRefPubMedPubMedCentralGoogle Scholar
  61. 61.
    Haraszti RA, Didiot MC, Sapp E, Leszyk J, Shaffer SA, Rockwell HE, et al. High-resolution proteomic and lipidomic analysis of exosomes and microvesicles from different cell sources. J Extracell Vesicles. 2016;5:32570.CrossRefPubMedGoogle Scholar
  62. 62.
    Hoshino A, Costa-Silva B, Shen TL, Rodrigues G, Hashimoto A, Tesic Mark M, et al. Tumour exosome integrins determine organotropic metastasis. Nature. 2015;527(7578):329–35.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2018

Authors and Affiliations

  • Scott Ferguson
    • 1
  • Sera Kim
    • 1
  • Christine Lee
    • 1
  • Michael Deci
    • 1
  • Juliane Nguyen
    • 1
  1. 1.Department of Pharmaceutical Sciences, School of PharmacyUniversity at Buffalo, The State University of New YorkBuffaloUSA

Personalised recommendations