Scleraxis Is Essential for Tendon Differentiation by Equine Embryonic Stem Cells and in Equine Fetal Tenocytes.

Bavin, EP
·
Atkinson, F
·
Barsby, T
·
Guest, DJ

Abstract: The transcription factor scleraxis is required for tendon development and is upregulated during embryonic stem cell (ESC) differentiation into tenocytes. However, its role beyond early embryonic development is not defined. We utilized a short hairpin RNA to knock down scleraxis expression in ESCs and adult and fetal tenocytes. No effect on growth or morphology was observed in two-dimensional cultures. However, scleraxis knockdown in fetal tenocytes significantly reduced COL1A1, COMP, and SOX9 gene expression. Scleraxis knockdown in adult tenocytes had no effect on the expression of these genes. Strikingly, differentiating ESCs and fetal tenocytes without scleraxis failed to reorganize a three-dimensional (3D) matrix and generate artificial tendons. This was associated with a significantly reduced survival. In contrast, there was no effect on the survival and remodeling capacity of adult tenocytes following scleraxis knockdown. Overexpression of scleraxis in fetal tenocytes rescued gene expression, cell survival in 3D, and subsequent matrix contraction. Together, these results demonstrate that scleraxis is not only essential for ESC differentiation into tenocytes but that it also has an active role in maintaining fetal tenocytes, which is then redundant in adult tenocytes.

Get Full Text

Publication Date: 2017-01-24 PubMed ID: 27899062DOI: 10.1089/scd.2016.0279Google 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.

Facebook X LinkedIn Email

Summary
Cite This
Publication
Affiliations
MeSH
Citations

Journal Article
Research Support
Non-U.S. Gov't

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 shows that a transcription factor named scleraxis, which is known to be necessary for tendon development and increases during embryonic stem cell differentiation into tenocytes, also plays an important role in maintaining fetal tendon cells. The knockdown of scleraxis results in the inability to generate artificial tendons, even though it does not impact the growth or shape of cells in standard cultures.

Objective, Design, and Methodology of the Research

The researchers used a technique known as short hairpin RNA (shRNA) to suppress the expression of the scleraxis transcription factor in embryonic stem cells (ESCs), and in adult and fetal tenocytes (tendon cells). This technique, known as silencing or ‘knockdown’, lets scientists explore the function of particular genes by reducing their activity.
The experiments were conducted in both two-dimensional cultures and three-dimensional (3D) matrices to evaluate the impact of scleraxis knockdown on cell behavior in both simple and complex environments. In the 3D matrix, the cells responded to the knockdown by reorganizing themselves and generating synthetic tendons.

Findings of the Research

Silencing scleraxis in ESCs and fetal tenocytes led to a significant reduction of specific gene expressions (COL1A1, COMP, and SOX9), essential for tendon development. But, silencing scleraxis in adult tenocytes did not affect the expression of these genes.
Remarkably, ESCs and fetal tenocytes without scleraxis were incapable of reorganizing in a 3D matrix and creating artificial tendons. They also exhibited a significantly reduced survival rate. However, adult tenocytes didn’t show any change in their survival and remodeling capacity post the scleraxis knockdown.
The researchers were able to reverse the effects of scleraxis suppression in fetal tenocytes by reintroducing scleraxis. This ‘rescue’ restored gene expression, cell survival in 3D, and subsequent matrix contraction.

Implications of the Research

The study thus implies that the scleraxis transcription factor is not only crucial for the differentiation of ESCs into tenocytes (tendon cells) but also plays a key role in maintaining fetal tenocytes.
Scleraxis’s role seems to become redundant in adult tenocytes, who can maintain themselves and remodel their structure even when its expression is inhibited.

Cite This Article

APA

Bavin EP, Atkinson F, Barsby T, Guest DJ. (2017). Scleraxis Is Essential for Tendon Differentiation by Equine Embryonic Stem Cells and in Equine Fetal Tenocytes. Stem Cells Dev, 26(6), 441-450. https://doi.org/10.1089/scd.2016.0279

Publication

Stem cells and development

ISSN: 1557-8534

NlmUniqueID: 101197107

Country: United States

Language: English

Volume: 26

Issue: 6

Pages: 441-450

Researcher Affiliations

Bavin, Emma P

1 Centre for Preventive Medicine , Animal Health Trust, Newmarket, United Kingdom .

Atkinson, Francesca

1 Centre for Preventive Medicine , Animal Health Trust, Newmarket, United Kingdom .

Barsby, Tom

1 Centre for Preventive Medicine , Animal Health Trust, Newmarket, United Kingdom .

Guest, Debbie J

1 Centre for Preventive Medicine , Animal Health Trust, Newmarket, United Kingdom .

MeSH Terms

Aging / physiology
Animals
Basic Helix-Loop-Helix Transcription Factors / genetics
Basic Helix-Loop-Helix Transcription Factors / metabolism
Cartilage Oligomeric Matrix Protein / metabolism
Cell Differentiation / drug effects
Cell Differentiation / genetics
Cell Proliferation / drug effects
Cell Survival / drug effects
Cells, Cultured
Collagen Type I / genetics
Collagen Type I / metabolism
DNA, Complementary / metabolism
Embryonic Stem Cells / cytology
Embryonic Stem Cells / drug effects
Embryonic Stem Cells / metabolism
Fetus / cytology
Gels / pharmacology
Gene Expression Regulation / drug effects
Gene Knockdown Techniques
Horses
RNA, Small Interfering / metabolism
SOX9 Transcription Factor / genetics
SOX9 Transcription Factor / metabolism
Tendons / cytology
Tenocytes / cytology
Tenocytes / drug effects
Tenocytes / metabolism

Citations

This article has been cited 14 times.

Weeratunga P, Harman RM, Van de Walle GR. Induced pluripotent stem cells from domesticated ruminants and their potential for enhancing livestock production.. Front Vet Sci 2023;10:1129287.
doi: 10.3389/fvets.2023.1129287pubmed: 36891466google scholar: lookup
Smith EJ, Beaumont RE, McClellan A, Sze C, Palomino Lago E, Hazelgrove L, Dudhia J, Smith RKW, Guest DJ. Tumour necrosis factor alpha, interleukin 1 beta and interferon gamma have detrimental effects on equine tenocytes that cannot be rescued by IL-1RA or mesenchymal stromal cell-derived factors.. Cell Tissue Res 2023 Mar;391(3):523-544.
doi: 10.1007/s00441-022-03726-6pubmed: 36543895google scholar: lookup
Papalamprou A, Yu V, Chen A, Stefanovic T, Kaneda G, Salehi K, Castaneda CM, Gertych A, Glaeser JD, Sheyn D. Directing iPSC differentiation into iTenocytes using combined scleraxis overexpression and cyclic loading.. J Orthop Res 2023 Jun;41(6):1148-1161.
doi: 10.1002/jor.25459pubmed: 36203346google scholar: lookup
Yang Q, Li J, Su W, Yu L, Li T, Wang Y, Zhang K, Wu Y, Wang L. Electrospun aligned poly(u03b5-caprolactone) nanofiber yarns guiding 3D organization of tendon stem/progenitor cells in tenogenic differentiation and tendon repair.. Front Bioeng Biotechnol 2022;10:960694.
doi: 10.3389/fbioe.2022.960694pubmed: 36110313google scholar: lookup
Shojaee A, Ejeian F, Parham A, Nasr Esfahani MH. Optimizing Tenogenic Differentiation of Equine Adipose-Derived Mesenchymal Stem Cells (eq-ASC) Using TGFB3 Along with BMP Antagonists.. Cell J 2022 Jul 27;24(7):370-379.
doi: 10.22074/cellj.2022.7892pubmed: 36043405google scholar: lookup
Best KT, Korcari A, Mora KE, Nichols AE, Muscat SN, Knapp E, Buckley MR, Loiselle AE. Scleraxis-lineage cell depletion improves tendon healing and disrupts adult tendon homeostasis.. Elife 2021 Jan 22;10.
doi: 10.7554/eLife.62203pubmed: 33480357google scholar: lookup
Scarfone RA, Pena SM, Russell KA, Betts DH, Koch TG. The use of induced pluripotent stem cells in domestic animals: a narrative review.. BMC Vet Res 2020 Dec 8;16(1):477.
doi: 10.1186/s12917-020-02696-7pubmed: 33292200google scholar: lookup
Paterson YZ, Cribbs A, Espenel M, Smith EJ, Henson FMD, Guest DJ. Genome-wide transcriptome analysis reveals equine embryonic stem cell-derived tenocytes resemble fetal, not adult tenocytes.. Stem Cell Res Ther 2020 May 19;11(1):184.
doi: 10.1186/s13287-020-01692-wpubmed: 32430075google scholar: lookup
Theodossiou SK, Schiele NR. Models of tendon development and injury.. BMC Biomed Eng 2019;1.
doi: 10.1186/s42490-019-0029-5pubmed: 32095779google scholar: lookup
Liao X, Falcon ND, Mohammed AA, Paterson YZ, Mayes AG, Guest DJ, Saeed A. Synthesis and Formulation of Four-Arm PolyDMAEA-siRNA Polyplex for Transient Downregulation of Collagen Type III Gene Expression in TGF-u03b21 Stimulated Tenocyte Culture.. ACS Omega 2020 Jan 28;5(3):1496-1505.
doi: 10.1021/acsomega.9b03216pubmed: 32010823google scholar: lookup
Shojaee A, Parham A. Strategies of tenogenic differentiation of equine stem cells for tendon repair: current status and challenges.. Stem Cell Res Ther 2019 Jun 18;10(1):181.
doi: 10.1186/s13287-019-1291-0pubmed: 31215490google scholar: lookup
Plachel F, Moroder P, Gehwolf R, Tempfer H, Wagner A, Auffarth A, Matis N, Pauly S, Tauber M, Traweger A. Risk Factors for Rotator Cuff Disease: An Experimental Study on Intact Human Subscapularis Tendons.. J Orthop Res 2020 Jan;38(1):182-191.
doi: 10.1002/jor.24385pubmed: 31161610google scholar: lookup
McClellan A, Evans R, Sze C, Kan S, Paterson Y, Guest D. A novel mechanism for the protection of embryonic stem cell derived tenocytes from inflammatory cytokine interleukin 1 beta.. Sci Rep 2019 Feb 26;9(1):2755.
doi: 10.1038/s41598-019-39370-4pubmed: 30808942google scholar: lookup
Nichols AEC, Settlage RE, Werre SR, Dahlgren LA. Novel roles for scleraxis in regulating adult tenocyte function.. BMC Cell Biol 2018 Aug 7;19(1):14.
doi: 10.1186/s12860-018-0166-zpubmed: 30086712google scholar: lookup