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Cell and Tissue Research

, Volume 368, Issue 1, pp 93–104 | Cite as

Downregulation of CITED2 contributes to TGFβ-mediated senescence of tendon-derived stem cells

  • Chao Hu
  • Yan Zhang
  • Kanglai Tang
  • Yuchun LuoEmail author
  • Yunpeng Liu
  • Wan Chen
Regular Article

Abstract

Tendon-derived stem cells (TDSCs) are multipotent adult stem cells with potential applications in tendon and tendon–bone junction repair. However, cellular characteristics change during in vitro passaging. Therefore, elucidation of the molecular and cellular mechanisms of tendon aging will be essential for the development of TDSC-based therapies. The aim of this study is to investigate the effect of CITED2, a nuclear regulator and transforming growth factor β2 (TGFβ2) on TDSC proliferation and senescence by comparing cells derived from Achilles tendon biopsies of young individuals (Y-TDSC) with those of older patients (O-TDSC). Our results showed that CITED2 mRNA and protein expression levels were significantly higher in Y-TDSCs than in O-TDSCs and O-TDSCs displayed decreased proliferation and increased senescence compared with Y-TDSCs. Furthermore, high levels of CITED2 protein expression in Y-TDSCs correlated with the downregulation of SP1 and p21 and the upregulation of MYC, potentially indicating the mechanism by which CITED2 upregulates TDSC proliferation. TGFβ2 was found to downregulate the expression of the CITED2 gene and knockdown of CITED2 abolished the effect of TGFβ2 on TDSC proliferation and senescence. Thus, the downregulation of CITED2 contributes to TGFβ-mediated senescence providing an insight into the molecular and cellular mechanisms that contribute to tendon aging and degeneration. Our findings may aid the development of cell-based therapies for tendon repair.

Keywords

Tendon-derived stem cells Senescence Cell proliferation Transforming growth factor β2 CITED2 

Notes

Acknowledgement

This study was supported by the Fund of Medical Research Foundation of Nanjing Military (ZD04).

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interest.

References

  1. Aggarwal S, Pittenger MF (2005) Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood 105:1815–1822CrossRefPubMedGoogle Scholar
  2. Ahmad Z, Wardale J, Brooks R, Henson F, Noorani A, Rushton N (2012) Exploring the application of stem cells in tendon repair and regeneration. Arthroscopy 28:1018–1029CrossRefPubMedGoogle Scholar
  3. Alberton P, Popov C, Pragert M, Kohler J, Shukunami C, Schieker M, Docheva D (2012) Conversion of human bone marrow-derived mesenchymal stem cells into tendon progenitor cells by ectopic expression of scleraxis. Stem Cells Dev 21:846–858CrossRefPubMedGoogle Scholar
  4. Alhadlaq A, Mao JJ (2004) Mesenchymal stem cells: isolation and therapeutics. Stem Cells Dev 13:436–448CrossRefPubMedGoogle Scholar
  5. Benjamin M, Kaiser E, Milz S (2008) Structure-function relationships in tendons: a review. J Anat 212:211–228CrossRefPubMedPubMedCentralGoogle Scholar
  6. Bi Y, Stuelten CH, Kilts T, Wadhwa S, Iozzo RV, Robey PG, Chen XD, Young MF (2005) Extracellular matrix proteoglycans control the fate of bone marrow stromal cells. J Biol Chem 280:30481–30489CrossRefPubMedGoogle Scholar
  7. Bi Y, Ehirchiou D, Kilts TM, Inkson CA, Embree MC, Sonoyama W, Li L, Leet AI, Seo BM, Zhang L, Shi S, Young MF (2007) Identification of tendon stem/progenitor cells and the role of the extracellular matrix in their niche. Nat Med 13:1219–1227CrossRefPubMedGoogle Scholar
  8. Bierie B, Moses HL (2006) Tumour microenvironment: TGFbeta: the molecular Jekyll and Hyde of cancer. Nat Rev Cancer 6:506–520CrossRefPubMedGoogle Scholar
  9. Bocker W, Rossmann O, Docheva D, Malterer G, Mutschler W, Schieker M (2007) Quantitative polymerase chain reaction as a reliable method to determine functional lentiviral titer after ex vivo gene transfer in human mesenchymal stem cells. J Gene Med 9:585–595CrossRefPubMedGoogle Scholar
  10. Brent AE, Schweitzer R, Tabin CJ (2003) A somitic compartment of tendon progenitors. Cell 113:235–248CrossRefPubMedGoogle Scholar
  11. Cashman JD, Eaves AC, Raines EW, Ross R, Eaves CJ (1990) Mechanisms that regulate the cell cycle status of very primitive hematopoietic cells in long-term human marrow cultures. I. Stimulatory role of a variety of mesenchymal cell activators and inhibitory role of TGF-beta. Blood 75:96–101PubMedGoogle Scholar
  12. Chou YT, Hsieh CH, Chiou SH, Hsu CF, Kao YR, Lee CC, Chung CH, Wang YH, Hsu HS, Pang ST, Shieh YS, Wu CW (2012) CITED2 functions as a molecular switch of cytokine-induced proliferation and quiescence. Cell Death Differ 19:2015–2028CrossRefPubMedPubMedCentralGoogle Scholar
  13. Docheva D, Padula D, Popov C, Weishaupt P, Pragert M, Miosge N, Hickel R, Bocker W, Clausen-Schaumann H, Schieker M (2010) Establishment of immortalized periodontal ligament progenitor cell line and its behavioural analysis on smooth and rough titanium surface. Eur Cell Mater 19:228–241CrossRefPubMedGoogle Scholar
  14. Finkel T, Serrano M, Blasco MA (2007) The common biology of cancer and ageing. Nature 448:767–774CrossRefPubMedGoogle Scholar
  15. Guerquin MJ, Charvet B, Nourissat G, Havis E, Ronsin O, Bonnin MA, Ruggiu M, Olivera-Martinez I, Robert N, Lu Y, Kadler KE, Baumberger T, Doursounian L, Berenbaum F, Duprez D (2013) Transcription factor EGR1 directs tendon differentiation and promotes tendon repair. J Clin Invest 123:3564–3576CrossRefPubMedPubMedCentralGoogle Scholar
  16. Hagedorn L, Floris J, Suter U, Sommer L (2000) Autonomic neurogenesis and apoptosis are alternative fates of progenitor cell communities induced by TGFbeta. Dev Biol 228:57–72CrossRefPubMedGoogle Scholar
  17. Huang GT, Shagramanova K, Chan SW (2006) Formation of odontoblast-like cells from cultured human dental pulp cells on dentin in vitro. J Endod 32:1066–1073CrossRefPubMedGoogle Scholar
  18. Ikushima H, Miyazono K (2012) TGF-beta signal transduction spreading to a wider field: a broad variety of mechanisms for context-dependent effects of TGF-beta. Cell Tissue Res 347:37–49CrossRefPubMedGoogle Scholar
  19. Kasper G, Mao L, Geissler S, Draycheva A, Trippens J, Kuhnisch J, Tschirschmann M, Kaspar K, Perka C, Duda GN, Klose J (2009) Insights into mesenchymal stem cell aging: involvement of antioxidant defense and actin cytoskeleton. Stem Cells 27:1288–1297CrossRefPubMedGoogle Scholar
  20. Kleczko EK, Kim J, Keysar SB, Heasley LR, Eagles JR, Simon M, Marshall ME, Singleton KR, Jimeno A, Tan AC, Heasley LE (2015) An inducible TGF-beta2-TGFbetaR pathway modulates the sensitivity of HNSCC cells to tyrosine kinase inhibitors targeting dominant receptor tyrosine kinases. PLoS One 10:e0123600CrossRefPubMedPubMedCentralGoogle Scholar
  21. Kohler J, Popov C, Klotz B, Alberton P, Prall WC, Haasters F, Muller-Deubert S, Ebert R, Klein-Hitpass L, Jakob F, Schieker M, Docheva D (2013) Uncovering the cellular and molecular changes in tendon stem/progenitor cells attributed to tendon aging and degeneration. Aging Cell 12:988–999CrossRefPubMedPubMedCentralGoogle Scholar
  22. Kranc KR, Oliveira DV, Armesilla-Diaz A, Pacheco-Leyva I, Catarina Matias A, Luisa Escapa A, Subramani C, Wheadon H, Trindade M, Nichols J, Kaji K, Enver T, Braganca J (2015) Acute loss of Cited2 impairs nanog expression and decreases self-renewal of mouse embryonic stem cells. Stem Cells 33:699–712CrossRefPubMedGoogle Scholar
  23. Laping NJ, Everitt JI, Frazier KS, Burgert M, Portis MJ, Cadacio C, Gold LI, Walker CL (2007) Tumor-specific efficacy of transforming growth factor-beta RI inhibition in eker rats. Clin Cancer Res 13:3087–3099CrossRefPubMedGoogle Scholar
  24. Liu L, Rando TA (2011) Manifestations and mechanisms of stem cell aging. J Cell Biol 193:257–266CrossRefPubMedPubMedCentralGoogle Scholar
  25. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−delta delta C(T)) method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  26. Mienaltowski MJ, Adams SM, Birk DE (2013) Regional differences in stem cell/progenitor cell populations from the mouse Achilles tendon. Tissue Eng Part A 19:199–210CrossRefPubMedGoogle Scholar
  27. Moses HL, Serra R (1996) Regulation of differentiation by TGF-beta. Curr Opin Genet Dev 6:581–586CrossRefPubMedGoogle Scholar
  28. Moses HL, Yang EY, Pietenpol JA (1990) TGF-beta stimulation and inhibition of cell proliferation: new mechanistic insights. Cell 63:245–247CrossRefPubMedGoogle Scholar
  29. Mueller MB, Fischer M, Zellner J, Berner A, Dienstknecht T, Prantl L, Kujat R, Nerlich M, Tuan RS, Angele P (2010) Hypertrophy in mesenchymal stem cell chondrogenesis: effect of TGF-beta isoforms and chondrogenic conditioning. Cells Tissues Organs 192:158–166CrossRefPubMedPubMedCentralGoogle Scholar
  30. Ni M, Lui PP, Rui YF, Lee YW, Tan Q, Wong YM, Kong SK, Lau PM, Li G, Chan KM (2012) Tendon-derived stem cells (TDSCs) promote tendon repair in a rat patellar tendon window defect model. J Orthop Res 30:613–619CrossRefPubMedGoogle Scholar
  31. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147CrossRefPubMedGoogle Scholar
  32. Qiu Y, Wang X, Zhang Y, Carr AJ, Zhu L, Xia Z, Sabokbar A (2014) Development of a refined tenocyte expansion culture technique for tendon tissue engineering. J Tissue Eng Regen Med 8:955–962CrossRefPubMedGoogle Scholar
  33. Rando TA (2006) Stem cells, ageing and the quest for immortality. Nature 441:1080–1086CrossRefPubMedGoogle Scholar
  34. Rees JD, Wilson AM, Wolman RL (2006) Current concepts in the management of tendon disorders. Rheumatology (Oxford) 45:508–521CrossRefGoogle Scholar
  35. Ryan JM, Barry FP, Murphy JM, Mahon BP (2005) Mesenchymal stem cells avoid allogeneic rejection. J Inflamm (Lond) 2:8CrossRefGoogle Scholar
  36. Sahin E, Depinho RA (2010) Linking functional decline of telomeres, mitochondria and stem cells during ageing. Nature 464:520–528CrossRefPubMedPubMedCentralGoogle Scholar
  37. Schuster N, Krieglstein K (2002) Mechanisms of TGF-beta-mediated apoptosis. Cell Tissue Res 307:1–14CrossRefPubMedGoogle Scholar
  38. Schweitzer R, Chyung JH, Murtaugh LC, Brent AE, Rosen V, Olson EN, Lassar A, Tabin CJ (2001) Analysis of the tendon cell fate using Scleraxis, a specific marker for tendons and ligaments. Development 128:3855–3866PubMedGoogle Scholar
  39. Sharpless NE, DePinho RA (2007) How stem cells age and why this makes us grow old. Nat Rev Mol Cell Biol 8:703–713CrossRefPubMedGoogle Scholar
  40. da Silva ML, Caplan AI, Nardi NB (2008) In search of the in vivo identity of mesenchymal stem cells. Stem Cells 26:2287–2299CrossRefGoogle Scholar
  41. Smith RK, Birch HL, Goodman S, Heinegard D, Goodship AE (2002) The influence of ageing and exercise on tendon growth and degeneration—hypotheses for the initiation and prevention of strain-induced tendinopathies. Comp Biochem Physiol A Mol Integr Physiol 133:1039–1050CrossRefPubMedGoogle Scholar
  42. Tempfer H, Wagner A, Gehwolf R, Lehner C, Tauber M, Resch H, Bauer HC (2009) Perivascular cells of the supraspinatus tendon express both tendon- and stem cell-related markers. Histochem Cell Biol 131:733–741CrossRefPubMedGoogle Scholar
  43. Tuite DJ, Renstrom PA, O’Brien M (1997) The aging tendon. Scand J Med Sci Sports 7:72–77CrossRefPubMedGoogle Scholar
  44. Zhou Z, Akinbiyi T, Xu L, Ramcharan M, Leong DJ, Ros SJ, Colvin AC, Schaffler MB, Majeska RJ, Flatow EL, Sun HB (2010) Tendon-derived stem/progenitor cell aging: defective self-renewal and altered fate. Aging Cell 9:911–915CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Chao Hu
    • 1
  • Yan Zhang
    • 1
  • Kanglai Tang
    • 2
  • Yuchun Luo
    • 1
    Email author
  • Yunpeng Liu
    • 1
  • Wan Chen
    • 2
  1. 1.Department of Orthopedics101st Hospital of PLAWuxiChina
  2. 2.Department of Orthopedics, Southwest HospitalThird Military Medical UniversityChongqingChina

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