Online first
Original paper
Published online: 2025-03-03

open access

Page views 393
Article views/downloads 172
Get Citation

Connect on Social Media

Connect on Social Media

Expression of O-GlcNAcylation in pulp tissue and DPSCs of healthy dental organs

María Cristina Franco-Arellanes1, Perla Xóchitl Toledo-Valdes2, Cynthia Díaz-Hernández1, Risk Díaz-Castillejos2, Eunice Daysi García-Reyes2, Saira Karina Ramírez-Thomé2, Beatriz Xóchitl Ávila-Curiel2, María Cristina Castañeda-Patlán1, Edgar Zenteno1, Carlos Josué Solórzano-Mata12

Abstract

Introduction. O-GlcNAcylation is a post-translational modification in which a single N-Acetyl-D-Glucosamine (GlcNAc) molecule is added to Ser or Thr residues of proteins. The O-N-acetylglucosaminyl transferase (OGT) enzyme is responsible for adding GlcNAc to the target proteins and N-acetyl-β-D-glucosaminidase (OGA) that removes the GlcNAc residue. O-GlcNAcylation has been described in the pathophysiology of several diseases; however, little has been studied in dental tissue. The aim of the present work is to characterise the product of O-GlcNAcylation and its enzymes at the tissue level in the dental pulp, as well as its expression in dental pulp stem cells (DPSCs) both in situ and in vitro. This enables the recognition of the behaviour of O-GlcNAcylation in pulp tissue without pathology.

Materials and methods. Pulp tissue was obtained from 10 healthy donors, and the expression of O-GlcNAc, OGT, and OGA was analysed using immunofluorescence with specific antibodies in different regions of the dental pulp. DPSCs were isolated, cultured, and identified with anti-STRO1 (antibody specific for human CD34+ cells, useful for DPSC identification). The expression of O-GlcNAc in DPSCs was confirmed in vitro through Western blot.

Results. Different regions of the dental pulp and DPSCs express O-GlcNAc and the enzymes OGT and OGA. O-GlcNAc and OGT expression was more prominent in the odontoblastic layer, cell-rich zone, and in the central core. OGA was distributed throughout the pulp tissue with lower immunoreactivity compared to OGT.

Conclusions. Our results suggest that O-GlcNAcylation may play a relevant role in human dental pulp homeostasis and in physiology of DPSCs.

Article available in PDF format

View PDF Download PDF file

References

  1. Yang X, Qian K. Protein O-GlcNAcylation: emerging mechanisms and functions. Nat Rev Mol Cell Biol. 2017; 18(7): 452–465.
  2. Lefebvre T, Issad T. 30 years old: O-GlcNAc reaches the age of reason — regulation of cell signaling and metabolism by o-glcnacylation. Front Endocrinol (Lausanne). 2015; 6: 17.
  3. Lazarus BD, Love DC, Hanover JA. Recombinant O-GlcNAc transferase isoforms: identification of O-GlcNAcase, yes tyrosine kinase, and tau as isoform-specific substrates. Glycobiology. 2006; 16(5): 415–421.
  4. Lewis BA. The role of O-GlcNAcylation in RNA polymerase II transcription. J Biol Chem. 2024; 300(3): 105705.
  5. Dupas T, Lauzier B, McGraw S. O-GlcNAcylation: the sweet side of epigenetics. Epigenetics Chromatin. 2023; 16(1): 49.
  6. Ciraku L, Esquea EM, Reginato MJ. O-GlcNAcylation regulation of cellular signaling in cancer. Cell Signal. 2022; 90: 110201.
  7. Issad T, Al-Mukh H, Bouaboud A, et al. Protein O-GlcNAcylation and the regulation of energy homeostasis: lessons from knock-out mouse models. J Biomed Sci. 2022; 29(1): 64.
  8. González UG, Castillo VJ, Ramírez JC, et al. O-GlcNAcilación y moléculas de adhesión en carcinomas. TIP Rev Esp Cienc Quím-Biol. 2024; 27: 1–13.
  9. Jiménez-Castillo V, Illescas-Barbosa D, Zenteno E, et al. Increased O-GlcNAcylation promotes IGF-1 receptor/PhosphatidyI Inositol-3 kinase/Akt pathway in cervical cancer cells. Sci Rep. 2022; 12(1): 4464.
  10. Xu Z, Isaji T, Fukuda T, et al. -GlcNAcylation regulates integrin-mediated cell adhesion and migration via formation of focal adhesion complexes. J Biol Chem. 2019; 294(9): 3117–3124.
  11. Seyrek K, Ivanisenko NV, König C, et al. Modulation of extrinsic apoptotic pathway by intracellular glycosylation. Trends Cell Biol. 2024; 34(9): 728–741.
  12. Loison I, Pioger A, Paget S, et al. OrgaRES Consortium. O-GlcNAcylation inhibition redirects the response of colon cancer cells to chemotherapy from senescence to apoptosis. Cell Death Dis. 2024; 15(10): 762.
  13. Wani WY, Chatham JC, Darley-Usmar V, et al. O-GlcNAcylation and neurodegeneration. Brain Res Bull. 2017; 133: 80–87.
  14. Lozano L, Guevara J, Lefebvre T, et al. Effect of amyloid-Β (25-35) in hyperglycemic and hyperinsulinemic rats, effects on phosphorylation and O-GlcNAcylation of tau protein. Neuropeptides. 2017; 63: 18–27.
  15. Chatham JC, Zhang J, Wende AR. Role of O-linked N-acetylglucosamine protein modification in cellular (patho)physiology. Physiol Rev. 2021; 101(2): 427–493.
  16. He XF, Hu X, Wen GJ, et al. O-GlcNAcylation in cancer development and immunotherapy. Cancer Lett. 2023; 566: 216258.
  17. Wu D, Cai Y, Jin J. Potential coordination role between O-GlcNAcylation and epigenetics. Protein Cell. 2017; 8(10): 713–723.
  18. Burt RA, Alghusen IM, John Ephrame S, et al. Mapping the O-GlcNAc modified proteome: applications for health and disease. Front Mol Biosci. 2022; 9: 920727.
  19. Yu C, Abbott PV. An overview of the dental pulp: its functions and responses to injury. Aust Dent J. 2007; 52(1 Suppl): S4–16.
  20. Gómez de Ferraris M, Campos A. Histología, embriología e ingeniería tisular bucodental. Médica Panamericana, Buenos Aires 2009.
  21. Fu X, Kim HS. Dentin mechanobiology: bridging the gap between architecture and function. Int J Mol Sci. 2024; 25(11).
  22. Yang Y, Zhao Y, Liu X, et al. Effect of SOX2 on odontoblast differentiation of dental pulp stem cells. Mol Med Rep. 2017; 16(6): 9659–9663.
  23. Sato M, Kawase-Koga Y, Yamakawa D, et al. Bone regeneration potential of human dental pulp stem cells derived from elderly patients and osteo-induced by a helioxanthin derivative. Int J Mol Sci. 2020; 21(20).
  24. Le Blanc K, Mougiakakos D. Multipotent mesenchymal stromal cells and the innate immune system. Nat Rev Immunol. 2012; 12(5): 383–396.
  25. Fernandes TL, Cortez de SantAnna JP, Frisene I, et al. Systematic review of human dental pulp stem cells for cartilage regeneration. Tissue Eng Part B Rev. 2020; 26(1): 1–12.
  26. Kim BC, Bae H, Kwon IK, et al. Osteoblastic/cementoblastic and neural differentiation of dental stem cells and their applications to tissue engineering and regenerative medicine. Tissue Eng Part B Rev. 2012; 18(3): 235–244.
  27. Lepperdinger G. Inflammation and mesenchymal stem cell aging. Curr Opin Immunol. 2011; 23(4): 518–524.
  28. Shi S, Gronthos S, Chen S, et al. Bone formation by human postnatal bone marrow stromal stem cells is enhanced by telomerase expression. Nat Biotechnol. 2002; 20(6): 587–591.
  29. Ning H, Lin G, Lue TF, et al. Mesenchymal stem cell marker Stro-1 is a 75 kd endothelial antigen. Biochem Biophys Res Commun. 2011; 413(2): 353–357.
  30. Yu J, He H, Tang C, et al. Differentiation potential of STRO-1+ dental pulp stem cells changes during cell passaging. BMC Cell Biol. 2010; 11: 32.
  31. Gross T, Dieterle MP, Vach K, et al. Biomechanical modulation of dental pulp stem cell (DPSC) properties for soft tissue engineering. Bioengineering (Basel). 2023; 10(3).
  32. Hu Y, You C, Song C, et al. The beneficial effect of global O-GlcNAcylation on odontogenic differentiation of human dental pulp cells via mTORC1 pathway. Arch Oral Biol. 2022; 138: 105427.
  33. Karamzadeh R, Eslaminejad MB, Aflatoonian R. Isolation, characterization and comparative differentiation of human dental pulp stem cells derived from permanent teeth by using two different methods. J Vis Exp. 2012(69).
  34. Shi S, Gronthos S. Perivascular niche of postnatal mesenchymal stem cells in human bone marrow and dental pulp. J Bone Miner Res. 2003; 18(4): 696–704.
  35. Baudoin L, Issad T. O-GlcNAcylation and inflammation: a vast territory to explore. Front Endocrinol (Lausanne). 2014; 5: 235.
  36. Ouchi T, Nakagawa T. Cellular signaling for dental physiological functions. Biomolecules. 2023; 13(8).
  37. Mamaladze MT, Ustiashvili MG. Theoretical and practical principles of dentinogenesis: hypotheses and confirmed clinically reality. Georgian Med News. 2010(186): 22–28.
  38. Hart G, Akimoto Y. The O-GlcNAc modification. In: Varki A, Cummings RD, Esko JD. ed. Essentials of glycobiology. 2nd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor (NY) 2009.
  39. Su C, Schwarz TL. O-GlcNAc transferase is essential for sensory neuron survival and maintenance. J Neurosci. 2017; 37(8): 2125–2136.
  40. Durand SH, Flacher V, Roméas A, et al. Lipoteichoic acid increases TLR and functional chemokine expression while reducing dentin formation in in vitro differentiated human odontoblasts. J Immunol. 2006; 176(5): 2880–2887.
  41. Pääkkönen V, Rusanen P, Hagström J, et al. Mature human odontoblasts express virus-recognizing toll-like receptors. Int Endod J. 2014; 47(10): 934–941.
  42. He W, Zhang Y, Zhang J, et al. Cytidine-phosphate-guanosine oligonucleotides induce interleukin-8 production through activation of TLR9, MyD88, NF-κB, and ERK pathways in odontoblast cells. J Endod. 2012; 38(6): 780–785.
  43. Dommisch H, Winter J, Açil Y, et al. Human beta-defensin (hBD-1, -2) expression in dental pulp. Oral Microbiol Immunol. 2005; 20(3): 163–166.
  44. Yang WHo, Park SY, Nam HW, et al. NFkappaB activation is associated with its O-GlcNAcylation state under hyperglycemic conditions. Proc Natl Acad Sci U S A. 2008; 105(45): 17345–17350.
  45. Very N, Boulet C, Gheeraert C, et al. O-GlcNAcylation controls pro-fibrotic transcriptional regulatory signaling in myofibroblasts. Cell Death Dis. 2024; 15(6): 391.
  46. Vang S, Helton ES, Guo Y, et al. O-GlcNAc transferase regulates collagen deposition and fibrosis resolution in idiopathic pulmonary fibrosis. Front Immunol. 2024; 15: 1387197.
  47. Masaki N, Feng B, Bretón-Romero R, et al. O-GlcNAcylation mediates glucose-induced alterations in endothelial cell phenotype in human diabetes mellitus. J Am Heart Assoc. 2020; 9(12): e014046.
  48. Lima VV, Giachini FR, Carneiro FS, et al. O-GlcNAcylation contributes to augmented vascular reactivity induced by endothelin 1. Hypertension. 2010; 55(1): 180–188.
  49. Wang Z, Park K, Comer F, et al. Site-specific GlcNAcylation of human erythrocyte proteins: potential biomarker(s) for diabetes. Diabetes. 2009; 58(2): 309–317.
  50. Park K, Saudek CD, Hart GW. Increased expression of beta-N-acetylglucosaminidase in erythrocytes from individuals with pre-diabetes and diabetes. Diabetes. 2010; 59(7): 1845–1850.
  51. Park K, Saudek CD, Hart GW. Increased expression of beta-N-acetylglucosaminidase in erythrocytes from individuals with pre-diabetes and diabetes. Diabetes. 2010; 59(7): 1845–1850.
  52. Oh M, Zhang Z, Mantesso A, et al. Endothelial-initiated crosstalk regulates dental pulp stem cell self-renewal. J Dent Res. 2020; 99(9): 1102–1111.
  53. Gronthos S, Mankani M, Brahim J, et al. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A. 2000; 97(25): 13625–13630.
  54. Shafi R, Iyer SP, Ellies LG, et al. The O-GlcNAc transferase gene resides on the X chromosome and is essential for embryonic stem cell viability and mouse ontogeny. Proc Natl Acad Sci U S A. 2000; 97(11): 5735–5739.
  55. Mitsiadis TA, Feki A, Papaccio G, et al. Dental pulp stem cells, niches, and notch signaling in tooth injury. Adv Dent Res. 2011; 23(3): 275–279.
  56. Lizier NF, Kerkis A, Gomes CM, et al. Scaling-up of dental pulp stem cells isolated from multiple niches. PLoS One. 2012; 7(6): e39885.
  57. Téclès O, Laurent P, Zygouritsas S, et al. Activation of human dental pulp progenitor/stem cells in response to odontoblast injury. Arch Oral Biol. 2005; 50(2): 103–108.