Therapeutic MSC exosomes are derived from lipid raft microdomains in the plasma membrane

Abstract

Background Mesenchymal stem cell (MSC) was previously shown to secrete lipid vesicles that when purified by high performance liquid chromatography as a population of homogenously sized particles with a hydrodynamic radius of 55–65 nm reduce infarct size in a mouse model of myocardial ischemia/reperfusion injury. As these vesicles exhibit many biophysical and biochemical properties of exosomes, they were identified as exosomes. Here we investigated if these lipid vesicles were indeed exosomes that have an endosomal biogenesis. Method In most cells, endocytosis is thought to occur at specialized microdomains known as lipid rafts. To demonstrate an endosomal origin for MSC exosomes, MSCs were pulsed with ligands e.g. transferrin (Tfs) and Cholera Toxin B (CTB) that bind receptors in lipid rafts. The endocytosed ligands were then chased to determine if they were incorporated into the exosomes. Results A fraction of exogenous Tfs was found to recycle into MSC exosomes. When MSCs were pulsed with labelled Tfs in the presence of chlorpromazine, an inhibitor of clathrin‐mediated endocytosis, Tf incorporation in CD81‐immunoprecipitate was reduced during the chase. CTB which binds GM1 gangliosides that are enriched in lipid rafts extracted exosome‐associated proteins, CD81, CD9, Alix and Tsg101 from MSC‐conditioned medium. Exogenous CTBs were pulse‐chased into secreted vesicles. Extraction of Tf‐ or CTB‐binding vesicles in an exosome preparation mutually depleted each other. Inhibition of sphingomyelinases reduced CTB‐binding vesicles. Conclusion Together, our data demonstrated that MSC exosomes are derived from endocytosed lipid rafts and that their protein cargo includes exosome‐associated proteins CD81, CD9, Alix and Tsg101.

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References 41

1. 1.

   Herzog EL, Chai L, Krause DS. Plasticity of marrow-derived stem cells. Blood. 2003;102:3483-93.

   Not in our library10.1182/blood-2003-05-1664

2. 2.

   Gronthos S, Franklin DM, Leddy HA, Robey PG, Storms RW, Gimble JM. Surface protein characterization of human adipose tissue-derived stromal cells. J Cell Physiol. 2001;189:54-63.

   Not in our library10.1002/jcp.1138

3. 3.

   Erices A, Conget P, Minguell JJ. Mesenchymal progenitor cells in human umbilical cord blood. Br J Haematol. 2000;109:235-42.

   Not in our library10.1046/j.1365-2141.2000.01986.x

4. 4.

   In't Anker PS, Scherjon SA, Kleijburg-van der Keur C, Noort WA, Claas FH, Willemze R, et al. Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation. Blood. 2003;102:1548-9.

   Not in our library10.1182/blood-2003-04-1291

5. 5.

   Kim J, Breunig MJ, Escalante LE, Bhatia N, Denu RA, Dollar BA. Biologic and immunomodulatory properties of mesenchymal stromal cells derived from human pancreatic islets. Cytotherapy. 2012;14:925-35.

   Not in our library10.3109/14653249.2012.684376

6. 6.

   Ricciardi M, Malpeli G, Bifari F, Bassi G, Pacelli L, Nwabo Kamdje AH, et al. Comparison of epithelial differentiation and immune regulatory properties of mesenchymal stromal cells derived from human lung and bone marrow. PLoS One. 2012;7:e35639.

   Not in our library10.1371/journal.pone.0035639

7. 7.

   Lian Q, Lye E, Suan Yeo K, Khia Way Tan E, Salto-Tellez M, Liu TM, et al. Derivation of clinically compliant MSCs from CD105+, CD24-differentiated human ESCs. Stem Cells. 2007;25:425-36.

   Not in our library10.1634/stemcells.2006-0420

8. 8.

   Lai RC, Choo A, Lim SK. Derivation and characterization of human ESC-derived mesenchymal stem cells. Methods Mol Biol. 2011;698:141-50.

   Not in our library10.1007/978-1-60761-999-4_11

9. 9.

   Jung Y, Bauer G, Nolta JA. Concise review: induced pluripotent stem cell-derived mesenchymal stem cells: progress toward safe clinical products. Stem Cells. 2012;30:42-7.

   Not in our library10.1002/stem.727

10. 10.

    Giordano A, Galderisi U, Marino IR. From the laboratory bench to the patient's bedside: an update on clinical trials with mesenchymal stem cells. J Cell Physiol. 2007;211:27-35.

    Not in our library10.1002/jcp.20959

11. 11.

    Bruder SP, Kurth AA, Shea M, Hayes WC, Jaiswal N, Kadiyala S. Bone regeneration by implantation of purified, culture-expanded human mesenchymal stem cells. J Orthop Res. 1998;16:155-62.

    Not in our library10.1002/jor.1100160202

12. 12.

    Johnstone B, Hering TM, Caplan AI, Goldberg VM, Yoo JU. In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res. 1998;238:265-72.

    Not in our library10.1006/excr.1997.3858

13. 13.

    Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284:143-7.

    Not in our library10.1126/science.284.5411.143

14. 14.

    Haynesworth SE, Goshima J, Goldberg VM, Caplan AI. Characterization of cells with osteogenic potential from human marrow. Bone. 1992;13:81-8.

    Not in our library10.1016/8756-3282(92)90364-3

15. 15.

    Yoo JU, Barthel TS, Nishimura K, Solchaga L, Caplan AI, Goldberg VM, et al. The chondrogenic potential of human bone-marrow-derived mesenchymal progenitor cells. J Bone Joint Surg Am. 1998;80:1745-57.

    Not in our library10.2106/00004623-199812000-00004

16. 16.

    Dennis JE, Merriam A, Awadallah A, Yoo JU, Johnstone B, Caplan AI. A quadripotential mesenchymal progenitor cell isolated from the marrow of an adult mouse. J Bone Miner Res. 1999;14:700-9.

    Not in our library10.1359/jbmr.1999.14.5.700

17. 17.

    Gojo S, Gojo N, Takeda Y, Mori T, Abe H, Kyo S, et al. In vivo cardiovasculogenesis by direct injection of isolated adult mesenchymal stem cells. Exp Cell Res. 2003;288:51-9.

    Not in our library10.1016/s0014-4827(03)00132-0

18. 18.

    Sanchez-Ramos J, Song S, Cardozo-Pelaez F, Hazzi C, Stedeford T, Willing A, et al. Adult bone marrow stromal cells differentiate into neural cells in vitro. Exp Neurol. 2000;164:247-56.

    Not in our library10.1006/exnr.2000.7389

19. 19.

    Woodbury D, Schwarz EJ, Prockop DJ, Black IB. Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res. 2000;61:364-70.

    Not in our library10.1002/1097-4547(20000815)61:4<364::aid-jnr2>3.0.co;2-c

20. 20.

    Kohyama J, Abe H, Shimazaki T, Koizumi A, Okano H, Hata J, et al. Brain from bone: efficient “meta-differentiation” of marrow stroma-derived mature osteoblasts to neurons with Noggin or a demethylating agent. Differentiation. 2001;68:235-44.

    Not in our library10.1046/j.1432-0436.2001.680411.x

21. 21.

    Kobayashi T, Hamano K, Li TS, Katoh T, Kobayashi S, Matsuzaki M, et al. Enhancement of angiogenesis by the implantation of self bone marrow cells in a rat ischemic heart model. J Surg Res. 2000;89:189-95.

    Not in our library10.1006/jsre.2000.5828

22. 22.

    Tomita S, Li RK, Weisel RD, Mickle DA, Kim EJ, Sakai T. Autologous transplantation of bone marrow cells improves damaged heart function. Circulation. 1999;100(19 Suppl):247-56.

    Not in our library10.1161/01.cir.100.suppl_2.ii-247

23. 23.

    Sato T, Iso Y, Uyama T, Kawachi K, Wakabayashi K, Omori Y, et al. Coronary vein infusion of multipotent stromal cells from bone marrow preserves cardiac function in swine ischemic cardiomyopathy via enhanced neovascularization. Lab Invest. 2011;91:553-64.

    Not in our library10.1038/labinvest.2010.202

24. 24.

    Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem. 2006;98:1076-84.

    Not in our library10.1002/jcb.20886

25. 25.

    Timmers L, Lim S-K, Arslan F, Armstrong JS, Hoefler IE, Doevendans PA, et al. Reduction of myocardial infarct size by human mesenchymal stem cell conditioned medium. Stem Cell Res. 2008;1:129-37.

    Not in our library10.1016/j.scr.2008.02.002

26. 26.

    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;4:214-22.

    Not in our library10.1016/j.scr.2009.12.003

27. 27.

    Chen TS, Lai RC, Lee MM, Choo AB, Lee CN, Lim SK. Mesenchymal stem cell secretes microparticles enriched in pre-microRNAs. Nucleic Acids Res. 2010;38:215-24.

    Not in our library10.1093/nar/gkp857

28. 28.

    Simons M, Raposo G. Exosomes – vesicular carriers for intercellular communication. Curr Opin Cell Biol. 2009;21:575-81.

    Not in our library10.1016/j.ceb.2009.03.007

29. 29.

    Zoller M. Tetraspanins: push and pull in suppressing and promoting metastasis. Nat Rev Cancer. 2009;9:40-55.

    Not in our library10.1038/nrc2543

30. 30.

    Fishman PH, Atikkan EE. Induction of cholera toxin receptors in cultured cells by butyric acid. J Biol Chem. 1979;254:4342-4.

    Not in our library

31. 31.

    Fujinaga Y, Wolf AA, Rodighiero C, Wheeler H, Tsai B, Allen L, et al. Gangliosides that associate with lipid rafts mediate transport of cholera and related toxins from the plasma membrane to endoplasmic reticulm. Mol Biol Cell. 2003;14:4783-93.

    Not in our library10.1091/mbc.e03-06-0354

32. 32.

    Laude AJ, Prior IA. Plasma membrane microdomains: organization, function and trafficking (Review). Mol Membr Biol. 2004;21:193-205.

    Not in our library10.1080/09687680410001700517

33. 33.

    Rajendran L, Simons K. Lipid rafts and membrane dynamics. J Cell Sci. 2005;118:1099-102.

    Not in our library10.1242/jcs.01681

34. 34.

    Trajkovic K, Hsu C, Chiantia S, Rajendran L, Wenzel D, Wieland F, et al. Ceramide triggers budding of exosome vesicles into multivesicular endosomes. Science. 2008;319:1244-7.

    Not in our library10.1126/science.1153124

35. 35.
    Enabling a robust scalable manufacturing process for therapeutic exosomes through oncogenic immortalization of human ESC-derived MSCs · 2011

    No notice
36. 36.

    Lai RC, Arslan F, Tan SS, Tan B, Choo A, Lee MM, et al. Derivation and characterization of human fetal MSCs: an alternative cell source for large-scale production of cardioprotective microparticles. J Mol Cell Cardiol. 2010;48:1215-24.

    Not in our library10.1016/j.yjmcc.2009.12.021

37. 37.

    Sze SK, de Kleijn DP, Lai RC, Tan EKW, Zhao H, Yeo KS, et al. Elucidating the secretion proteome of human embryonic stem cell-derived mesenchymal stem cells. Mol Cell Proteomics. 2007;6:1680-9.

    Not in our library10.1074/mcp.m600393-mcp200

38. 38.
    Proteolytic Potential of the MSC Exosome Proteome: Implications for an Exosome-Mediated Delivery of Therapeutic Proteasome · 2012

    No notice
39. 39.

    Lai R, Yeo R, Tan S, Zhang B, Yin Y, Sze S, LGV Chase, Mohan C. Mesenchymal stem cell exosomes: the future MSC-based therapy? Mesenchymal stem cell therapy. 2013;39-62.

    Not in our library

40. 40.

    Holmgren J, Lonnroth I, Mansson J, Svennerholm L. Interaction of cholera toxin and membrane GM1 ganglioside of small intestine. Proc Natl Acad Sci U S A. 1975;72:2520-4.

    Not in our library10.1073/pnas.72.7.2520

41. 41.

    Trajkovic K, Hsu C, Chiantia S, Rajendran L, Wenzel D, Wieland F, et al. Ceramide triggers budding of exosome vesicles into multivesicular endosomes. Science. 2008;319:1244-7.

    Not in our library10.1126/science.1153124

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