Analyze Diet
International journal of molecular sciences2025; 26(4); 1784; doi: 10.3390/ijms26041784

Role of Glycans in Equine Endometrial Cell Uptake of Extracellular Vesicles Derived from Amniotic Mesenchymal Stromal Cells.

Abstract: Extracellular vesicles (EVs) are important mediators of cell-cell communication thanks to their ability to transfer their bioactive cargo, thus regulating a variety of physiological contexts. EVs derived from amniotic mesenchymal/stromal cells (eAMC-EVs) are internalized by equine endometrial cells (eECs) with positive effects on regenerative medicine treatments. As the cellular uptake of EVs is influenced by the glycan profile of both EVs and target cells, this study is focused on the role of surface glycans in the uptake of eAMC-EVs by recipient eECs. Equine ECs were obtained by enzymatic digestion of uteri from healthy mares. Equine AMC-EVs were isolated from amniotic cell cultures according to a standardized protocol. The glycan pattern was studied using a panel of lectins in combination with fucosidase and neuraminidase treatment. Both eECs and eAMC-EVs expressed N-linked high mannose glycans, as well as fucosylated and sialylated glycans. All these glycans were involved in the uptake of eAMC-EVs by eECs. The internalization of eAMC-EVs was strongly reduced after cleavage of α1,2-linked fucose and α2,3/α2,6-linked sialic acids. These results demonstrate that surface glycans are involved in the internalization of eAMC-EVs by eECs and that fucosylated and sialylated glycans are highly relevant in the transfer of bioactive molecules with effects on regenerative medicine treatments.
Publication Date: 2025-02-19 PubMed ID: 40004247PubMed Central: PMC11855797DOI: 10.3390/ijms26041784Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates how surface sugars, or glycans, play a role in the absorption of extracellular vesicles (EVs) from amniotic mesenchymal stromal cells by equine endometrial cells. The study finds that certain types of glycans strongly influence this process and thus may impact treatments in regenerative medicine.

Study Objective and Methodology

  • The purpose of the study was to understand how surface sugars or glycans are involved in the absorption of extracellular vesicles (eAMC-EVs) from amniotic mesenchymal/stromal cells by equine endometrial cells (eECs). Glycan profiles can influence cellular uptake of EVs so this study focused on the role of glycans in this interaction.
  • The researchers collected equine endometrial cells from healthy horse wombs using an enzymatic digestion process. They isolated eAMC-EVs from horse amniotic cell cultures following a standard protocol.
  • They then studied the sugar pattern using a panel of lectins, sugar-binding proteins, combined with fucosidase and neuraminidase treatment. This method enabled them to identify the types of glycans present on both the eECs and the eAMC-EVs.

Results and Findings

  • The researchers discovered that N-linked high mannose glycans, fucosylated and sialylated glycans were expressed by both the equine endometrial cells (eECs) and amniotic mesenchymal/stromal cell-derived extracellular vesicles (eAMC-EVs).
  • All these glycans were involved in the uptake of eAMC-EVs by eECs, confirming the vital role glycans play in cellular uptake of EVs.
  • Further, the internalization process of eAMC-EVs was significantly reduced after the cleavage of α1,2-linked fucose and α2,3/α2,6-linked sialic acids, thereby confirming the importance of fucosylated and sialylated glycans in the process.

Conclusion and Implications

  • The study concludes that surface glycans, specifically fucosylated and sialylated glycans, are crucial for the internalization of eAMC-EVs by eECs.
  • This finding is highly significant for regenerative medicine treatments, as extracellular vesicles are known to carry bioactive molecules that can regulate physiological processes. Thus, understanding the role of glycans in the uptake of these vesicles can help improve the effectiveness of such treatments.

Cite This Article

APA
Gaspari G, Lange-Consiglio A, Cremonesi F, Desantis S. (2025). Role of Glycans in Equine Endometrial Cell Uptake of Extracellular Vesicles Derived from Amniotic Mesenchymal Stromal Cells. Int J Mol Sci, 26(4), 1784. https://doi.org/10.3390/ijms26041784

Publication

ISSN: 1422-0067
NlmUniqueID: 101092791
Country: Switzerland
Language: English
Volume: 26
Issue: 4
PII: 1784

Researcher Affiliations

Gaspari, Giulia
  • Laboratory of Reproduction and Regenerative Medicine, Department of Veterinary Medicine and Animal Science (DIVAS), University of Milan, 26900 Lodi, Italy.
Lange-Consiglio, Anna
  • Laboratory of Reproduction and Regenerative Medicine, Department of Veterinary Medicine and Animal Science (DIVAS), University of Milan, 26900 Lodi, Italy.
Cremonesi, Fausto
  • Laboratory of Reproduction and Regenerative Medicine, Department of Veterinary Medicine and Animal Science (DIVAS), University of Milan, 26900 Lodi, Italy.
Desantis, Salvatore
  • Department of Precision and Regenerative Medicine and Ionian Area, University of Bari Aldo Moro, 70010 Valenzano, Italy.

MeSH Terms

  • Animals
  • Extracellular Vesicles / metabolism
  • Horses
  • Female
  • Endometrium / metabolism
  • Endometrium / cytology
  • Mesenchymal Stem Cells / metabolism
  • Mesenchymal Stem Cells / cytology
  • Polysaccharides / metabolism
  • Amnion / metabolism
  • Amnion / cytology
  • Cells, Cultured
  • Fucose / metabolism

Grant Funding

  • PRIN202223FCREM_01 / Ministero dell'Istruzione, dell'Universitu00e0 e della Ricerca (MUR), Bando PRIN 2022 Settore LS9

Conflict of Interest Statement

The authors declare no conflicts of interest.

References

This article includes 49 references
  1. Arévalo-Turrubiarte M, Baratta M, Ponti G, Chiaradia E, Martignani E. Extracellular vesicles from equine mesenchymal stem cells decrease inflammation markers in chondrocytes in vitro.. Equine Vet. J. 2022;54:1133–1143.
    doi: 10.1111/evj.13537pmc: PMC9787580pubmed: 34741769google scholar: lookup
  2. Soukup R, Gerner I, Gültekin S, Baik H, Oesterreicher J, Grillari J, Jenner F. Characterisation of extracellular vesicles from equine mesenchymal stem cells.. Int. J. Mol. Sci. 2022;23:5858.
    doi: 10.3390/ijms23105858pmc: PMC9145091pubmed: 35628667google scholar: lookup
  3. Lange Consiglio A, Perrini C, Tasquier R, Deregibus M.C, Camussi G, Pascucci L, Marini M.G, Corradetti B, Bizzaro D, De Vita B. Microvesicles secreted from equine amniotic-derived cells and their potential role in reducing inflammation in a tendon in vitro model.. Stem Cells Dev. 2016;25:610–621.
    doi: 10.1089/scd.2015.0348pubmed: 26914245google scholar: lookup
  4. Perrini C, Strillacci M.G, Bagnato A, Esposti P, Marini M.G, Corradetti B, Bizzarro D, Idda A, Ledda S, Capra E. Microvesicles secreted from equine amniotic-derived cells and their potential role in reducing inflammation in endometrial cells in an in-vitro model.. Stem Cell Res. Ther. 2016;7:169.
    doi: 10.1186/s13287-016-0429-6pmc: PMC5114748pubmed: 27863532google scholar: lookup
  5. Lange-Consiglio A, Funghi F, Cantile C, Idda A, Cremonesi F, Riccaboni P. Case Report: Use of amniotic microvesicles for regenerative medicine treatment of a mare with chronic endometritis.. Front. Vet. Sci. 2020;7:347.
    doi: 10.3389/fvets.2020.00347pmc: PMC7311574pubmed: 32626730google scholar: lookup
  6. Varki A, Gagneux P. Biological functions of glycans.. In: Varki A., editor. Essentials of Glycobiology. 3rd ed. Cold Spring Harbor Laboratory Press; New York, NY, USA: 2017. pp. 77–88.
  7. Walker S.A, Aguilar Diaz De Leon J.S, Busatto S, Wurtz G, Zubair A.C, Borges C.R, Wolfram J. Glycan node analysis of plasma-derived extracellular vesicles.. Cells 2020;9:1946.
    doi: 10.3390/cells9091946pmc: PMC7563425pubmed: 32842648google scholar: lookup
  8. Macedo-da-Silva J, Santiago V.F, Rosa-Fernandes L, Marinho C.R.F, Palmisano G. Protein glycosylation in extracellular vesicles: Structural characterization and biological functions.. Mol. Immunol. 2021;135:226–246.
    doi: 10.1016/j.molimm.2021.04.017pubmed: 33933815google scholar: lookup
  9. Islam K, Khan M, Gidwani K, Witwer K.W, Lamminmaki U, Leivo J. Lectins as potential tools for cancer biomarker discovery from extracellular vesicles.. Biomark. Res. 2023;11:85.
    doi: 10.1186/s40364-023-00520-6pmc: PMC10540341pubmed: 37773167google scholar: lookup
  10. Williams C, Royo F, Aizpurua-Olaizola O, Pazos R, Boons G.J, Reichardt N.-C, Falcon-Perez J.M. Glycosylation of extracellular vesicles: Current knowledge, tools and clinical perspectives.. J. Extracell. Vesicles. 2018;7:1442985.
  11. Williams C, Pazos R, Royo F, González E, Meritxell R.-F, Aitor M, Gamiz J, Christian N, Reichardt N.-C, Falcón-Pérez J.M. Assessing the role of surface glycans of extracellular vesicles on cellular uptake.. Sci. Rep. 2019;9:11920.
    doi: 10.1038/s41598-019-48499-1pmc: PMC6695415pubmed: 31417177google scholar: lookup
  12. Li B, Hao K, Ma C, Li Z, Li H, Du W, Sun L, Jia T, Liu A, Li Y. Isolation and characterization of fucosylated extracellular vesicles based on a novel high-throughput GlyExo-Capture technique.. BioRxiv 2021.
    doi: 10.1101/2021.12.09.471505google scholar: lookup
  13. Zheng W, He R, Liang X, Roudi S, Bost J, Coly P.-M, van Guillaume N, Andaloussi S.E.L. Cell-specific targeting of extracellular vesicles though engineering the glycocalyx.. J. Extracell. Vesicles. 2022;11:12290.
    doi: 10.1002/jev2.12290pmc: PMC9719568pubmed: 36463392google scholar: lookup
  14. Shimoda A, Miura R, Tateno H, Seo N, Shiku H, Sawada S.I, Sasaki Y, Akiyoshi K. Assessment of surface glycan diversity on extracellular vesicles by lectin microarray and glycoengineering strategies for drug delivery applications.. Small Methods 2022;6:2100785.
    doi: 10.1002/smtd.202100785pubmed: 35174988google scholar: lookup
  15. Shimoda A, Tahara Y, Sawada S.-I, Sasaki Y, Akiyoshi K. Glycan profiling analysis using evanescent-field fluorescence assisted lectin array: Importance of sugar recognition for cellular uptake of exosomes from mesenchymal stem cells.. Biochem. Biophys. Res. Commun. 2017;491:701–707.
    doi: 10.1016/j.bbrc.2017.07.126pubmed: 28751214google scholar: lookup
  16. Royo F, Cossío U, Ruiz De Angulo A, Llop J, Falcon-Perez J.M. Modification of the glycosylation of extracellular vesicles alters their biodistribution in mice.. Nanoscale 2019;11:1531–1537.
    doi: 10.1039/C8NR03900Cpubmed: 30623961google scholar: lookup
  17. Sharon N, Lis H. History of lectins: From hemagglutinins to biological recognition molecules.. Glycobiology 2004;14:53R–62R.
    doi: 10.1093/glycob/cwh122pubmed: 15229195google scholar: lookup
  18. Brooks S.A. Lectins as versatile tools to explore cellular glycosylation.. Eur. J. Histochem. 2024;68:3959.
    doi: 10.4081/ejh.2024.3959pmc: PMC11059468pubmed: 38285057google scholar: lookup
  19. Krishnamoorthy L, Bess Jr J.W, Preston A.B, Nagashima K, Mahal L.K. HIV-1 and microvesicles from T cells share a common glycome, arguing for a common origin.. Nat. Chem. Biol. 2009;5:244–250.
    doi: 10.1038/nchembio.151pmc: PMC2713040pubmed: 19234452google scholar: lookup
  20. Batista B.S, Eng W.S, Pilobello K.T, Hendricks-Munoz K.D, Mahal L.K. Identification of a conserved glycan signature for microvesicles.. J. Proteome Res. 2011;10:4624–4633.
    doi: 10.1021/pr200434ypmc: PMC3443565pubmed: 21859146google scholar: lookup
  21. Desantis S, Accogli G, Albrizio M, Rossi R, Cremonesi F, Lange-Consiglio A. Glycan Profiling Analysis of equine amniotic progenitor mesenchymal cells and their derived extracellular microvesicles.. Stem Cells Dev. 2019;28:812–821.
    doi: 10.1089/scd.2019.0013pubmed: 30900531google scholar: lookup
  22. Slivka E.V, Shilova N.V, Obraztsova E.A, Kapustkina D.S, Khaidukov S.V, Nokel A.Y, Ryzhov I.M, Henry S.M, Bovin N.V, Rapoport E.M. Surface glycans of microvesicles derived from endothelial cells, as probed using plant lectins.. Int. J. Mol. Sci. 2024;25:5725.
    doi: 10.3390/ijms25115725pmc: PMC11171894pubmed: 38891913google scholar: lookup
  23. Lange-Consiglio A, Gaspari G, Funghi F, Capra E, Cretich M, Frigerio R, Bosi G, Cremonesi F. Amniotic mesenchymal-derived extracellular vesicles and their role in the prevention of persistent post-breeding induced endometritis.. Int. J. Mol. Sci. 2023;24:5166.
    doi: 10.3390/ijms24065166pmc: PMC10049450pubmed: 36982240google scholar: lookup
  24. Théry C, Witwer K.W, Aikawa E, Alcaraz M.J, Anderson J.D, Andriantsitohaina R, Antoniou A, Arab T, Archer F, Atkin-Smith G.K. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): A position statement of the international society for extracellular vesicles and update of the MISEV2014 Guidelines.. J. Extracell. Vesicles. 2018;7:1535750.
  25. Lange Consiglio A, Lazzari B, Perrini C, Pizzi F, Stella A, Cremonesi F, Capra E. MicroRNAs of equine amniotic mesenchymal cells-derived microvesicles and their involvement in anti-inflammatory process.. Cell Transplant. 2018;27:45–54.
    doi: 10.1177/0963689717724796pmc: PMC6434479pubmed: 29562776google scholar: lookup
  26. Lis H, Sharon N. The Antigens.. Volume IV. Academic Press; Cambridge, MA, USA: 1977. Lectins: Their chemistry and application to immunology; pp. 429–529.
  27. Goldstein I.J, Hayes C.E. The Lectins: Carbohydrate-binding proteins of plants and animals.. Adv. Carbohydr. Chem. Biochem. 1978;35:127–340.
    doi: 10.1016/s0065-2318(08)60220-6pubmed: 356549google scholar: lookup
  28. Kennedy J.F, Palva P.M.G, Corella M.T.S, Cavalcanti M.S.M, Coelho L.C.B.B. Lectins, versatile proteins of recognition: A review.. Carbohydr. Polym. 1995;26:219–230.
  29. Bojar D, Meche L, Meng G, Eng W, Smith D, Cummings R.D, Mahal L.K. A Useful guide to lectin binding: Machine-learning directed annotation of 57 unique lectin specificities.. ACS Chem. Biol. 2022;17:2993–3012.
    doi: 10.1021/acschembio.1c00689pmc: PMC9679999pubmed: 35084820google scholar: lookup
  30. Escrevente C, Keller S, Altevogt P, Costa J. Interaction and uptake of exosomes by ovarian cancer cells.. BMC Cancer 2011;11:108.
    doi: 10.1186/1471-2407-11-108pmc: PMC3072949pubmed: 21439085google scholar: lookup
  31. Terävä J, Verhassel A, Botti O, Islam M.K, Leivo J, Wittfooth S, Härkönen P, Pettersson K, Gidwani K. Primary breast cancer biomarkers based on glycosylation and extracellular vesicles detected from human serum.. Cancer Rep. 2021;5:1540.
    doi: 10.1002/cnr2.1540pmc: PMC9351655pubmed: 34423573google scholar: lookup
  32. Schwarz F, Aebi M. Mechanisms and principles of N-linked protein glycosylation.. Curr. Opin. Struct. Biol. 2011;21:576–582.
    doi: 10.1016/j.sbi.2011.08.005pubmed: 21978957google scholar: lookup
  33. Mathivanan S, Simpson R.J. ExoCarta: A compendium of exosomal proteins and RNA.. Proteomics 2009;9:4997–5000.
    doi: 10.1002/pmic.200900351pubmed: 19810033google scholar: lookup
  34. Paolini L, Orizio F, Busatto S, Radeghieri A, Bresciani R, Bergese P, Monti E. Exosomes secreted by Hela cells shuttle on their surface the plasma membrane-associated sialidase NEU3.. Biochemistry 2017;56:6401–6408.
    doi: 10.1021/acs.biochem.7b00665pubmed: 29039925google scholar: lookup
  35. Varki A, Schnaar R.L, Schauer R. Sialic acids and other nonulosonic acids.. In: Varki A., editor. Essentials of Glycobiology. 3rd ed. Cold Spring Harbor Laboratory Press; New York, NY, USA: 2017. pp. 179–195.
  36. Hamouda H, Ullah M, Berger M, Sittinger M, Tauber R, Ringe J, Blanchard V. N-glycosylation profile of undifferentiated and adipogenically differentiated human bone marrow mesenchymal stem cells: Towards a next generation of stem cell markers.. Stem Cells Dev. 2013;2:3100–3113.
    doi: 10.1089/scd.2013.0108pmc: PMC3856714pubmed: 23829188google scholar: lookup
  37. Lange-Consiglio A, Accogli G, Cremonesi F, Desantis S. Cell surface glycan changes in the spontaneous epithelial-mesenchymal transition of equine amniotic multipotent progenitor cells.. Cells Tissues Organs 2015;200:212–226.
    doi: 10.1159/000433420pubmed: 26337136google scholar: lookup
  38. Schauer R. Achievements and challenges of sialic acid research.. Glycoconj. J. 2000;17:485–499.
    doi: 10.1023/A:1011062223612pmc: PMC7087979pubmed: 11421344google scholar: lookup
  39. Zhao Y.Y, Takahashi M, Gu J.G, Miyoshi E, Matsumoto A, Kitazume S, Taniguchi N. Functional roles of N-glycans in cell signaling and cell adhesion in cancer.. Cancer Sci. 2008;99:1304–1310.
  40. Weidemann W, Klukas C, Klein A, Simm A, Schreiber F, Horstkorte R. Lessons from GNE-deficient embryonic stem cells: Sialic acid biosynthesis is involved in proliferation and gene expression.. Glycobiology 2010;20:107–117.
    doi: 10.1093/glycob/cwp153pubmed: 19797319google scholar: lookup
  41. Kang J, Park H.M, Kim Y.W, Kim Y.H, Varghese S, Seok H.K, Kim Y.G, Kim S.H. Control of mesenchymal stem cell phenotype and differentiation depending on cell adhesion mechanism.. Eur. Cell Mater. 2014;28:387–403.
    doi: 10.22203/eCM.v028a27pubmed: 25422949google scholar: lookup
  42. Staudacher E, Altmann F, Wilson I.B, März L. Fucose in N-glycans: From plant to man.. Biochem. Biophys. Acta. 1999;1473:216–236.
    doi: 10.1016/S0304-4165(99)00181-6pubmed: 10580141google scholar: lookup
  43. Becker D.J, Lowe J.B. Fucose: Biosynthesis and biological function in mammals.. Glycobiology 2003;13:41R–53R.
    doi: 10.1093/glycob/cwg054pubmed: 12651883google scholar: lookup
  44. Dai Z, Liu Y.K, Cui J.F, Shen H.L, Chen J, Sun R.X, Zhang Y, Zhou X.W, Yang P.Y, Tang Z.Y. Identification and analysis of altered α1, 6-fucosylated glycoproteins associated with hepatocellular carcinoma metastasis.. Proteomics 2006;6:5857–5867.
    doi: 10.1002/pmic.200500707pubmed: 17068759google scholar: lookup
  45. Aldi S, Della Giovampaola C, Focarelli R, Armini A, Ziche M, Finetti F, Rosati F. A fucose-containing O-glycoepitope on bovine and human nucleolin.. Glycobiology 2009;19:337–343.
    doi: 10.1093/glycob/cwn126pubmed: 19023103google scholar: lookup
  46. Zentella R, Wang Y, Zahn E, Hu J, Jiang L, Hunt D.F, Sun T.-P. Spindly O-fucosylates nuclear and cytoplasmic proteins involved in diverse cellular processes in plants.. Plant Physiol. 2023;191:1546–1560.
    doi: 10.1093/plphys/kiad011pmc: PMC10022643pubmed: 36740243google scholar: lookup
  47. Zhu W, Zhou Y, Guo L, Feng S. Biological function of sialic acid and sialylation in human health and disease.. Cell Death Discov. 2024;10:415.
    doi: 10.1038/s41420-024-02180-3pmc: PMC11442784pubmed: 39349440google scholar: lookup
  48. Chen X, Xiao S, Wu J, Yao J. Effect of sialic acid on mammalian cell culture and protein expression: A potential productivity enhancer for biopharmaceutical cell culture processes.. Processes 2020;8:1449.
    doi: 10.3390/pr8111449google scholar: lookup
  49. Alisson-Silva F, de Carvalho Rodrigues D, Vairo L, Asensi K.D, Vasconcelos-dos-Santos A, Mantuano N.R, Dias W.B, Rondinelli E, Goldenberg R.C, Urmenyi T.P. Evidences for the involvement of cell surface glycans in stem cell pluripotency and differentiation.. Glycobiology 2014;24:458–468.
    doi: 10.1093/glycob/cwu012pubmed: 24578376google scholar: lookup

Citations

This article has been cited 0 times.