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Cited by 15Bone Metabolism and DiseasesWnt/β-catenin signaling in development and cancerMesenchymal stem cell research

The DOI matches its Crossref record; 39 of the 39 references it cites were found in a registry or an index; 1 of the 22 sources its claims cite were checked for support.

N-myc downstream regulated gene 1 suppresses osteoblast differentiation through inactivating Wnt/β-catenin signaling

Open access
Shi, Xiaoli, Cen, Yunzhu, Shan, Liying, Tian, Lijie, Zhu, Endong, Yuan, Hairui, Li, Xiaoxia, Liu, Ying, Wang, Baoli·2022·Stem Cell Research & Therapy
DOI 10.1186/s13287-022-02714-5PMC PMC8817551
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Abstract

Background: N-myc downstream regulated gene 1 (NDRG1) plays a role in a variety of biological processes including differentiation of osteoclasts. However, it is not known if and how NDRG1 regulates osteogenic differentiation of marrow stromal progenitor cells. Methods: Gene expression profiling analysis was performed to study the expression level of Ndrg1 during osteogenic and adipogenic differentiation. Gain-of-function and/or loss-of function experiments were carried out to study the role of NDRG1 in the proliferation and differentiation of marrow stromal progenitor cells and the mechanism underlying the function was investigated. Finally, in vivo transfection of Ndrg1 siRNA was done and its effect on osteogenic and adipogenic differentiation in mice was explored. Results: Gene expression profiling analysis revealed that NDRG1 level was regulated during osteogenic and adipogenic differentiation of progenitor cells. The functional experiments demonstrated that NDRG1 negatively regulated the cell growth, and reciprocally modulated the osteogenic and adipogenic commitment of marrow stromal progenitor cells, driving the cells to differentiate toward adipocytes at the expense of osteoblast differentiation. Moreover, NDRG1 interacted with low-density lipoprotein receptor-related protein 6 (LRP6) in the stromal progenitor cells and inactivated the canonical Wnt/β-catenin signaling cascade. Furthermore, the impaired differentiation of progenitor cells induced by Ndrg1 siRNA could be attenuated when β-catenin was simultaneously silenced. Finally, in vivo transfection of Ndrg1 siRNA to the marrow of mice prevented the inactivation of canonical Wnt signaling in the BMSCs of ovariectomized mice, and ameliorated the reduction of osteoblasts on the trabeculae and increase of fat accumulation in the marrow observed in the ovariectomized mice. Conclusion: This study has provided evidences that NDRG1 plays a role in reciprocally modulating osteogenic and adipogenic commitment of marrow stromal progenitor cells through inactivating canonical Wnt signaling.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

NDRG1 is regulated during osteogenic/adipogenic differentiation and acts as an anti-osteogenic, pro-adipogenic factor in marrow stromal progenitor cells. It does so by interacting with LRP6, destabilizing it, and inactivating canonical Wnt/β-catenin signaling.

Key findings

  • NDRG1 expression was high in kidney and bone, changed during osteogenic and adipogenic differentiation, and was increased in BMSCs from ovariectomized mice and in calvaria from 18-month-old versus 3-month-old mice.
  • NDRG1 overexpression blocked osteogenic differentiation, decreasing osteogenic factor mRNAs by 45–68% and proteins by 42–56%; Ndrg1 knockdown enhanced osteogenesis, increasing mRNAs by 61–83% and proteins by 49–65%.
  • NDRG1 overexpression promoted adipogenic differentiation, increasing oil-red O retention by 49%, adipogenic mRNAs by 1.7–2.4 fold, and proteins by 45–72%; knockdown reduced oil-red O retention by 33%, mRNAs by 37–75%, and proteins by 36–61%.
  • NDRG1 interacted with LRP6, reduced phospho-LRP6(S1490), phospho-GSK3β(S9), non-phospho-β-catenin, and TCF7L2, and destabilized LRP6; β-catenin silencing attenuated the pro-osteogenic/anti-adipogenic effects of Ndrg1 siRNA. In vivo, Ndrg1 siRNA in ovariectomized mice increased osteocalcin-positive osteoblasts and reduced marrow adipocyte number and area.

Methods

  • Gain- and loss-of-function studies used primary mouse BMSCs and ST2 cells with Ndrg1 overexpression constructs, siRNA, or shRNA lentiviruses; differentiation was assessed by ALP/alizarin red staining, oil-red O staining, qRT-PCR, and Western blotting.
  • Mechanistic assays included cellular immunofluorescence for β-catenin, co-immunoprecipitation for NDRG1–LRP6 interaction, and cycloheximide treatment for LRP6 stability.
  • In vivo, C57BL/6 mice received intra-tibial 2′-Ome Ndrg1 siRNA or control siRNA; sham and ovariectomized groups were analyzed for BMSC Wnt/β-catenin proteins, osteogenic/adipogenic differentiation, and tibial histology/immunohistochemistry. Statistics used mean ± SD, t tests, and ANOVA with p < 0.05.

Limitations

  • Not stated in the paper.
  • Evident from the text: findings rely on mouse BMSCs, ST2 cells, and female ovariectomized mice, with no human validation.
  • Evident from the text: in vivo NDRG1 loss-of-function uses siRNA knockdown rather than genetic deletion, and the LRP6 degradation mechanism is inferred from stability assays without direct ubiquitination/proteasome experiments.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1186/s13287-022-02714-5

Cites

39 of 39 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

1 of 22 cited sources checked · 1 supported · 0 flagged

Verify all 9 references — free account
  • Findingown data

    Ndrg1 expression is upregulated at days 5 through 14 in ST2 cells during osteogenic differentiation.

    “For ST2 cells, Ndrg1 was upregulated at day 5 through day 14 (Fig. 1C).”
    Quote in the paperNot a substantial claim
  • Findingown data

    Ndrg1 depletion promotes osteogenic differentiation of ST2 cells.

    “Moreover, Ndrg1 depletion also promoted osteogenic differentiation of ST2 cells (Additional file 2: Fig. S1E–H).”
    Quote in the paperSubstantial claim
  • Findingown data

    Overexpressing Ndrg1 in ST2 cells promotes adipogenic differentiation.

    “Additionally, overexpressing Ndrg1 in ST2 cells promoted the adipogenic differentiation as well (Additional file 2: Fig. S2A–D).”
    Quote in the paperNot a substantial claim
  • Findingown data

    Ndrg1 overexpression alleviates osteogenic differentiation of ST2 cells.

    “Moreover, Ndrg1 overexpression also alleviated osteogenic differentiation of ST2 cells (Additional file 2: Fig. S1A–D).”
    Quote in the paperSubstantial claim
  • Findingown data

    Ndrg1 expression is increased in BMSCs from ovariectomized mice versus sham-operated mice.

    “Moreover, mRNA expression assay revealed that Ndrg1 was increased in the BMSCs from ovariectomized (OVX) mice versus those from sham-operated mice (Fig. 1F) or in the calvaria from 18-month-old aged mice versus 3-month-old young mice (Fig. 1G).”
    Quote in the paperNot a substantial claim
  • Probably not a substantial claim. 12
  • It may state two separate claims as one. 19

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    NDRG1 expression is changed in osteocytes of rats after treatment with sclerostin antibody.

    “A recent study reported the changed expression of NDRG1 in osteocytes of rats after treatment with sclerostin antibody [25].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [25] Taylor S, Hu R, Pacheco E, Locher K, Pyrah I, Ominsky MS, et al. Differential time-dependent transcriptional changes in the osteoblast lineage in cortical bone associated with sclerostin antibody treatment in ovariectomized rats. Bone Rep. 2018;8:95–103. source

  • It may state two separate claims as one.

    Findingown data

    Ndrg1 expression is downregulated at day 2 and upregulated at days 8 through 14 in primary BMSCs during osteogenic differentiation.

    “It was shown by using qRT-PCR that the expression of Ndrg1 was downregulated at day 2, but upregulated at day 8 through 14 in primary BMSCs during osteogenic differentiation (Fig. 1B).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Knockdown of Ndrg1 stimulates osteogenic differentiation in primary BMSCs, enhancing ALP and alizarin red staining and increasing mRNA/protein levels of osteogenic factors (mRNAs increased by 61-83%, proteins by 49-65%).

    “Following osteogenic treatment, knockdown of Ndrg1 stimulated osteogenic differentiation, as indicated by the enhanced ALP staining and alizarin red staining (Fig. 2F), and the increased mRNA and protein levels of the osteogenic factors as compared to the infection of the control virus (mRNAs increased by 61–83% and proteins increased by 49–65%) (Fig. 2G, H).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Ndrg1 expression is downregulated at day 1 and upregulated at days 3 and/or 5 during adipogenic differentiation.

    “By contrast, Ndrg1 was downregulated at the early stage (day 1) and upregulated at later stage (day 3 and/or day 5) of induction during adipogenic differentiation (Fig. 1D, E).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Findingown data

    Ndrg1 overexpression suppresses the cell growth rate of ST2 cells by 17%.

    “Furthermore, Ndrg1 overexpression suppressed the cell growth rate of ST2 cells by 17% (Fig. S1I).”
    Quote in the paperNot a substantial claim

  • It may state two separate claims as one.

    Findingown data

    Overexpressing Ndrg1 in primary BMSCs stimulates adipogenic differentiation, increasing oil-red O retention by 49% and mRNA/protein levels of PPARγ, C/EBPα, FABP4, and adipsin (mRNAs increased by 1.7-2.4 fold, proteins by 45-72%).

    “For the gain-of-function experiments, after adipogenic treatment, overexpressing Ndrg1 in primary BMSCs stimulated adipogenic differentiation, which was evidenced by the enhanced oil-red O staining (49% increase of oil-red O retention in the cells) (Fig. 3A, B) and the induced mRNA and protein levels of the adipogenic factors such as PPARγ, C/EBPα, FABP4 and adipsin as compared to the infection of”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    NDRG1 directly interacts with LRP6 and reduces phosphorylation of LRP6 in stromal cells.

    “Consistent to this, we also demonstrated that NDRG1 directly interacted with LRP6 and reduced the phosphorylation of LRP6 in stromal cells.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    In ST2 cells, NDRG1 overexpression reduces the protein levels of phospho-LRP6(S1490), phospho-GSK3β(S9), non-phospho-β-catenin, and TCF7L2.

    “Western blotting analysis showed that the overexpression of NDRG1 significantly reduced the protein levels of phospho-LRP6(S1490), phospho-GSK3β(S9), non-phospho-β-catenin and transcription factor 7 like 2 (TCF7L2).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Downregulation of NDRG1 in the marrow of mice increases osteoblasts and decreases adipocytes in either wild-type or ovariectomized mice.

    “Of more importance, the physiological role of NDRG1 was validated as the downregulation of NDRG1 in the marrow of mice was able to increase osteoblasts and decrease adipocytes in either the wild-type mice or the ovariectomized mice.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Ndrg1 siRNA specifically knocks down Ndrg1 without off-target effects.

  • It may state two separate claims as one.

    Findingown data

    Under adipogenic conditions, Ndrg1 siRNA reduces oil-red O staining and decreases levels of adipogenic factors compared to control siRNA.

    “By contrast, under adipogenic condition, the cells cotransfected with Ndrg1 siRNA and control siRNA displayed alleviated oil-red O staining and decreased levels of adipogenic factors as compared to the cells transfected with control siRNA only.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Ndrg1 siRNA specifically knocks down Ndrg1 without off-target effects.

  • It may state two separate claims as one.

    Findingown data

    Under osteogenic conditions, Ndrg1 siRNA enhances ALP staining and increases levels of osteogenic factors compared to control siRNA in marrow stromal cells.

    “Under osteogenic condition, the cells cotransfected with Ndrg1 siRNA and control siRNA displayed enhanced ALP staining and increased levels of osteogenic factors as compared to the cells transfected with control siRNA only.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Ndrg1 siRNA specifically knocks down Ndrg1 without off-target effects.

  • It may state two separate claims as one.

    Findingown data

    In ST2 cells, Ndrg1 overexpression inhibits and Ndrg1 depletion promotes the translocation of β-catenin from the cytoplasm into the nucleus.

    “It revealed that the dysregulation of NDRG1 changed the intracellular distribution of β-catenin. Briefly, Ndrg1 overexpression inhibited, while Ndrg1 depletion promoted the translocation of β-catenin from the cytoplasm into the nucleus (Fig. 4A, B).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    In ST2 cells, Ndrg1 depletion increases the protein levels of phospho-LRP6(S1490), phospho-GSK3β(S9), non-phospho-β-catenin, and TCF7L2.

    “Conversely, these proteins were increased following depletion of Ndrg1 (Fig. 4C, D).”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Findingown data

    Downregulating Ndrg1 in primary BMSCs blunts adipogenic differentiation, decreasing oil-red O retention by 33% and mRNA/protein levels of adipogenic factors (mRNAs decreased by 37-75%, proteins by 36-61%).

    “For the loss-of-function experiments, after adipogenic treatment, downregulating Ndrg1 in primary BMSCs blunted adipogenic differentiation, which was evidenced by the attenuated oil-red O staining (33% decrease of oil-red O retention in the cells) (Fig. 3E, F) and the decreased mRNA and protein levels of the adipogenic factors as compared to the infection of the control virus (mRNAs decreased by 3”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Phosphorylation of NDRG1 results in increased levels of N-cadherin, α-catenin, and β-catenin.

    “The phosphorylation of NDRG1 results in increased levels of the main adhesion molecules such as N-cadherin, α-catenin, and β-catenin [13].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [13] Schonkeren SL, Massen M, van der Horst R, Koch A, Vaes N, Melotte V. Nervous NDRGs: the N-myc downstream-regulated gene family in the central and peripheral nervous system. Neurogenetics. 2019;20:173–86. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    The best characterized regulators of osteogenesis include Wnt/β-catenin pathway, BMPs, Runx2, and osterix.

    “The best characterized regulators of osteogenesis include Wnt/β-catenin pathway, bone morphogenetic proteins (BMPs), runt related transcription factor-2 (Runx2), and osterix (OSX), etc. [3–7].”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [3] Wu M, Chen G, Li YP. TGF-beta and BMP signaling in osteoblast, skeletal development, and bone formation, homeostasis and disease. Bone Res. 2016;4:16009. source
    • cites [4] Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR, et al. The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell. 2002;108:17–29. source
    • cites [5] Alman BA. The role of hedgehog signalling in skeletal health and disease. Nat Rev Rheumatol. 2015;11:552–60. source
    • cites [6] Salazar VS, Gamer LW, Rosen V. BMP signalling in skeletal development, disease and repair. Nat Rev Endocrinol. 2016;12:203–21. source
    • cites [7] Lerner UH, Ohlsson C. The WNT system: background and its role in bone. J Intern Med. 2015;277:630–49. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    NDRG1 is implicated in the differentiation of osteoclasts and bone remodeling.

    “Watari et al. have recently demonstrated the implication of NDRG1 in the differentiation of osteoclasts and bone remodeling [24].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [24] Watari K, Shibata T, Nabeshima H, Shinoda A, Fukunaga Y, Kawahara A, et al. Impaired differentiation of macrophage lineage cells attenuates bone remodeling and inflammatory angiogenesis in Ndrg1 deficient mice. Sci Rep. 2016;6:19470. source

  • It may state two separate claims as one.

    Interpretationown data

    NDRG1 has an anti-osteogenic and pro-adipogenic function in mesenchymal progenitor cells.

    “The data uncovered the anti-osteogenic and pro-adipogenic function of NDRG1 in mesenchymal progenitor cells.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationown data

    NDRG1 may play a role in osteogenic and/or adipogenic differentiation of progenitor cells.

    “These data suggest that NDRG1 may play a role in osteogenic and/or adipogenic differentiation of progenitor cells.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    A reciprocal and competitive fate decision is frequently observed between adipogenesis and osteogenesis.

    “A reciprocal and competitive fate decision is frequently observed between adipogenesis and osteogenesis [1, 2].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [1] Chen Q, Shou P, Zheng C, Jiang M, Cao G, Yang Q, et al. Fate decision of mesenchymal stem cells: Adipocytes or osteoblasts? Cell Death Differ. 2016;23:1128–39. source
    • cites [2] Veldhuis-Vlug AG, Rosen CJ. Mechanisms of marrow adiposity and its implications for skeletal health. Metabolism. 2017;67:106–14. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    PPARγ and C/EBPs are the major transcription factors controlling adipocyte differentiation.

    “In contrast, peroxisome proliferator activated receptor γ (PPARγ) and CCAAT/enhancer binding proteins (C/EBPs) are the major transcription factors controlling adipocyte differentiation [8, 9].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [8] de Mota SP, Richard AJ, Hang H, Stephens JM. Transcriptional regulation of adipogenesis. Compr Physiol. 2017;7:635–74.

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites [9] Sadie-Van Gijsen H, Hough FS, Ferris WF. Determinants of bone marrow adiposity: the modulation of peroxisome proliferator-activated receptor-gamma2 activity as a central mechanism. Bone. 2013;56:255–65. source

  • It may state two separate claims as one.

    Premisecitation

    NDRG1 is capable of interacting with LRP6 and preventing its phosphorylation by Wnt ligands, thereby blocking de-phosphorylated inactivation of GSK3β and subsequent activation of Wnt signaling.

    “It was reported that NDRG1 is capable of interacting with LRP6 and preventing its phosphorylation by Wnt ligands, thereby blocking de-phosphorylated inactivation of GSK3β and subsequent activation of Wnt signaling [16].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [16] Liu W, Xing F, Iiizumi-Gairani M, Okuda H, Watabe M, Pai SK, et al. N-myc downstream regulated gene 1 modulates Wnt-beta-catenin signalling and pleiotropically suppresses metastasis. EMBO Mol Med. 2012;4:93–108. source

  • It may state two separate claims as one.

    Interpretationown data

    NDRG1 suppresses β-catenin translocation from the cytoplasm to the nucleus and downregulates active non-phospho-β-catenin and TCF7L2, demonstrating its inhibitory effect on the canonical Wnt pathway.

    “In line with our hypothesis, NDRG1 suppressed the translocation of β-catenin from the cytoplasm to the nucleus, and downregulated the protein levels of active non-phospho-β-catenin and TCF7L2, demonstrating its inhibitory effect on canonical Wnt pathway.”
    Quote in the paperSubstantial claim

  • It may state two separate claims as one.

    Interpretationown data

    NDRG1 suppresses osteogenesis and promotes adipogenesis via interacting with LRP6 to inactivate the Wnt/β-catenin signaling cascade.

    “This supports the hypothesis that NDRG1 suppresses osteogenesis and conversely promotes adipogenesis via interacting with LRP6 to inactivate Wnt/β-catenin signaling cascade.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: β-catenin siRNA effectively depletes β-catenin in the cotransfection experiments.
    • Rests on an assumption: Ndrg1 siRNA specifically knocks down Ndrg1 without off-target effects.

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Study design and conductnone given

    Female mice aged 8 weeks were randomly allocated into four groups.

    “the female mice aged 8 weeks were randomly allocated into four groups”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim.

    Premisecitation

    Wnt-induced phosphorylation of LRP6 at five reiterated PPPSPxS motifs and adjacent Ser/Thr clusters is a key step in Wnt signaling.

    “A key step in this procedure is the Wnt-induced phosphorylation of of LRP6 at five reiterated PPPSPxS motifs and adjacent Ser/Thr clusters [38].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [38] Tamai K, Zeng X, Liu C, Zhang X, Harada Y, Chang Z, et al. A mechanism for Wnt coreceptor activation. Mol Cell. 2004;13:149–56. source

  • Probably not a substantial claim. The paper’s conclusions may not depend on it.

    Premisecitation

    Canonical Wnt signaling is involved as a major mechanism in the control of osteogenesis and bone mass accrual by many regulators such as BMPs, NFI family members, and microRNAs.

    “Canonical Wnt signaling is involved as a major mechanism in the control of osteogenesis and bone mass accrual by many regulators such as BMPs, nuclear factor-I (NFI) family members and microRNAs [31–36].”
    Quote in the paperNot a substantial claim0 of 6 sources checked
    • cites [31] Kamiya N, Kobayashi T, Mochida Y, Yu PB, Yamauchi M, Kronenberg HM, et al. Wnt inhibitors Dkk1 and Sost are downstream targets of BMP signaling through the type IA receptor (BMPRIA) in osteoblasts. J Bone Miner Res. 2010;25:200–10. source
    • cites [32] Zhou J, Wang S, Qi Q, Yang X, Zhu E, Yuan H, et al. Nuclear factor I–C reciprocally regulates adipocyte and osteoblast differentiation via control of canonical Wnt signaling. FASEB J. 2017;31:1939–52. source
    • cites [33] Wu X, Wang X, Shan L, Zhou J, Zhang X, Zhu E, et al. High-mobility group AT-Hook 1 mediates the role of nuclear factor I/X in osteogenic differentiation through activating canonical Wnt signaling. Stem Cells. 2021;38:499–513. source
    • cites [34] Feng L, Zhang JF, Shi L, Yang ZM, Wu TY, Wang HX, et al. MicroRNA-378 suppressed Osteogenesis of MSCs and impaired bone formation via inactivating Wnt/beta-Catenin signaling. Mol Ther Nucleic Acids. 2020;21:1017–28. source
    • cites [35] Zhang L, Tang Y, Zhu X, Tu T, Sui L, Han Q, et al. Overexpression of MiR-335-5p promotes bone formation and regeneration in mice. J Bone Miner Res. 2017;32:2466–75. source
    • cites [36] Garcia J, Smith SS, Karki S, Drissi H, Hrdlicka HH, Youngstrom DW, et al. miR-433-3p suppresses bone formation and mRNAs critical for osteoblast function in mice. J Bone Miner Res. 2021. source

  • Probably not a substantial claim.

    Premisecitation

    LRP6, one of the co-receptors of Wnt ligands, is required for the transduction of the Wnt/β-catenin signaling cascade.

    “It is commonly known that LRP6, one of the co-receptors of Wnt ligands, is required for the transduction of Wnt/β-catenin signaling cascade.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [37] Schupbach D, Comeau-Gauthier M, Harvey E, Merle G. Wnt modulation in bone healing. Bone. 2020;138:115491. source

  • It may state two separate claims as one.

    Interpretationreasoning

    NDRG1 interacts with LRP6, blocks canonical Wnt signaling, and impairs the differentiation of marrow stromal progenitor cells.

    “Mechanistic investigations suggest that NDRG1 interacts with the Wnt co-receptor, LRP6, thereby blocks the canonical Wnt signaling, and thus orchestrates a cellular network that impairs the differentiation of marrow stromal progenitor cells.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim. It may state two separate claims as one.

    Premisecitation

    Activation of canonical Wnt signaling increases bone mass and strength, while inhibition of the pathway decreases bone mass and strength.

    “The activation of the canonical Wnt signaling usually increases, while the inhibition of the pathway decreases bone mass and strength [30].”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [30] Baron R, Gori F. Targeting WNT signaling in the treatment of osteoporosis. Curr Opin Pharmacol. 2018;40:134–41. source
30 of 30

  • The paper relies on this without support.

    The exact effect of NDRG1 on osteoblast differentiation has not been reported.

    “However, the exact effect of NDRG1 on osteoblast differentiation has not been reported.”

  • The authors state this as an assumption.

    It is not known if and how NDRG1 plays a role in the cell fate decision of BMSCs.

    “However, it is not known if and how NDRG1 plays a role in the cell fate decision of BMSCs.”

  • The paper relies on this without support.

    β-catenin siRNA effectively depletes β-catenin in the cotransfection experiments.

    “This pro-osteogenic effect of Ndrg1 siRNA was attenuated when the cells were cotransfected with Ndrg1 siRNA and β-catenin siRNA (Fig. 5A–C).”

  • The paper relies on this without support.

    Ndrg1 siRNA specifically knocks down Ndrg1 without off-target effects.

    “qRT-PCR analysis validated the efficacy of Ndrg1 siRNA in knocking down the expression of endogenous Ndrg1 in the BMSCs of tibiae (Fig. 6A).”
4 of 4

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Figure 5 citation omits the adipogenic protein panel

    Minor

    The results cite Fig. 5D-F for the decreased levels of adipogenic factors, but the figure caption assigns their protein measurements to panel G. The cited range therefore omits one of the relevant panels.

1 of 1

Cited works that were retracted

Works this paper cites that were later retracted, from the retraction notices publishers deposit. Major when the notice came out before this paper was published.

  • Cites a retracted work: 10.1016/j.bbrc.2018.10.054

    Info

    The cited work was retracted on 2022-06-13 (notice 10.1016/j.bbrc.2022.05.097), not before this paper was published (2022-02-04).

    Cited work 10.1016/j.bbrc.2018.10.054 · retracted 2022-06-13 · notice 10.1016/j.bbrc.2022.05.097 · this paper published 2022-02-04

1 of 1

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Metadata

Authors
Shi, X., Cen, Y., Shan, L., Tian, L., Zhu, E., Yuan, H., Li, X., Liu, Y., & Wang, B.
Affiliations
Tianjin Medical University (CN)
Corresponding author
  • Li, Xiaoxia
  • Liu, Ying
  • Wang, Baoli
ORCID
Venue
Stem Cell Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2022-02-04
Volume
13
Issue
1
Pages
53-53
ISSN
1757-6512 (ISSN-L)
DOI
10.1186/s13287-022-02714-5
PMC
PMC8817551
OpenAlex
W4210336749
Cited by
15
References
39
Citations by year
2026: 3 · 2025: 4 · 2024: 6 · 2023: 2
FWCI
0.88
Type
journal article
Language
en
Open access
Licence
Locations
  • Stem Cell Research & Therapy (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • national natural science foundation of china (82072389, 81871741, 82002264, 81972033, 81972031)
  • natural science foundation of tianjin city (18JCZDJC32200)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Mesenchymal stem cell research
Keywords
NDRG1, osteogenic differentiation, Wnt/β-catenin signaling, adipogenesis, LRP6, ovariectomy, bone loss, osteoblasts
MeSH terms
N-myc Downstream-Regulated Gene 1 Protein, Animals, Cell Differentiation, Osteoblasts, Osteogenesis, Cell Cycle Proteins, Intracellular Signaling Peptides and Proteins, beta Catenin, Mice, Wnt Signaling Pathway

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A language model drew the claims from the paper on 2026-11-10, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-11-10 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

Cited sources are checked with a language model’s help (gpt-6-sol).

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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