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Arthritis Res Ther
2013 Sep 19;155:R126. doi: 10.1186/ar4306.
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GREM1, FRZB and DKK1 mRNA levels correlate with osteoarthritis and are regulated by osteoarthritis-associated factors.
Leijten JC
,
Bos SD
,
Landman EB
,
Georgi N
,
Jahr H
,
Meulenbelt I
,
Post JN
,
van Blitterswijk CA
,
Karperien M
.
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INTRODUCTION: Osteoarthritis is, at least in a subset of patients, associated with hypertrophic differentiation of articular chondrocytes. Recently, we identified the bone morphogenetic protein (BMP) and wingless-type MMTV integration site (WNT) signaling antagonists Gremlin 1 (GREM1), frizzled-related protein (FRZB) and dickkopf 1 homolog (Xenopus laevis) (DKK1) as articular cartilage's natural brakes of hypertrophic differentiation. In this study, we investigated whether factors implicated in osteoarthritis or regulation of chondrocyte hypertrophy influence GREM1, FRZB and DKK1 expression levels.
METHODS: GREM1, FRZB and DKK1 mRNA levels were studied in articular cartilage from healthy preadolescents and healthy adults as well as in preserved and degrading osteoarthritic cartilage from the same osteoarthritic joint by quantitative PCR. Subsequently, we exposed human articular chondrocytes to WNT, BMP, IL-1β, Indian hedgehog, parathyroid hormone-related peptide, mechanical loading, different medium tonicities or distinct oxygen levels and investigated GREM1, FRZB and DKK1 expression levels using a time-course analysis.
RESULTS: GREM1, FRZB and DKK1 mRNA expression were strongly decreased in osteoarthritis. Moreover, this downregulation is stronger in degrading cartilage compared with macroscopically preserved cartilage from the same osteoarthritic joint. WNT, BMP, IL-1β signaling and mechanical loading regulated GREM1, FRZB and DKK1 mRNA levels. Indian hedgehog, parathyroid hormone-related peptide and tonicity influenced the mRNA levels of at least one antagonist, while oxygen levels did not demonstrate any statistically significant effect. Interestingly, BMP and WNT signaling upregulated the expression of each other's antagonists.
CONCLUSIONS: Together, the current study demonstrates an inverse correlation between osteoarthritis and GREM1, FRZB and DKK1 gene expression in cartilage and provides insight into the underlying transcriptional regulation. Furthermore, we show that BMP and WNT signaling are linked in a negative feedback loop, which might prove essential in articular cartilage homeostasis by balancing BMP and WNT activity.
Figure 1. GREM1, FRZB and DKK1 expression in healthy cartilage and osteoarthritic cartilage. Relative GREM1, FRZB and DKK1 mRNA expression levels of paired specimens of macroscopically preserved and degenerating osteoarthritic cartilage from a single osteoarthritic joint (n = 23) were assessed by quantitative PCR and were compared with healthy preadolescent (n = 4) and healthy adult (n = 3) articular cartilage specimen. Data expressed as fold-change relative to the specimen with the mRNA expression level on a log scale. *P <0.05. DKK1, dickkopf 1 homolog (Xenopus laevis); FRZB, frizzled-related protein; GADPH, glyceraldehyde 3-phosphate dehydrogenase; GREM1, Gremlin 1; OA, osteoarthritis.
Figure 2. Effects of bone morphogenetic protein signaling on the mRNA expression of GREM1, FRZB and DKK1. Chondrocytes were stimulated for 48 hours with different concentrations of BMP2 ranging from 0 to 200 ng/μl. Effects on gene expression of GREM1, FRZB, DKK1, AXIN2 and DKK1 were analyzed using quantitative PCR. Data expressed as fold-change relative to control and represents the mean of three donors ± standard deviation. *P <0.05 compared with 0 ng/ml BMP2. BMP2, bone morphogenetic protein 2; DKK1, dickkopf 1 homolog (Xenopus laevis); FRZB, frizzled-related protein; GADPH, glyceraldehyde 3-phosphate dehydrogenase; GREM1, Gremlin 1.
Figure 3. Effect of canonical WNT signaling on the mRNA expression of GREM1, FRZB and DKK1. (A) Primary human chondrocytes were exposed to 100 ng/ml WNT3A or three different concentrations of GIN. After 48 hours, AXIN2 mRNA expression was analyzed by quantitative PCR. (B) to (F) Chondrocytes were exposed to a single dose of 10 nM GIN or 100 ng/ml WNT3A. At indicated time points, mRNA expression was analyzed by quantitative PCR of AXIN2(B), FRZB(C), DKK1(D), GREM1(E) and ID1(F). Data expressed as fold-change relative to untreated time-point-matched control and represents the mean of three donors ± standard deviation. *P <0.05 compared with unstimulated cells (A) or 0 hours of stimulation (B) to (F). DKK1, dickkopf 1 homolog (Xenopus laevis); FRZB, frizzled-related protein; GADPH, glyceraldehyde 3-phosphate dehydrogenase; GREM1, Gremlin 1; WNT, wingless-type MMTV integration site.
Figure 4. Effects of IL-1β-mediated signaling on mRNA expression of GREM1, FRZB and DKK1. Chondrocytes received a single dose of 10 or 100 ng/ml IL-1β, or a daily medium refreshment containing 10 ng/ml IL-1β. Chondrocytes were stimulated up to 96 hours and mRNA expression was analyzed by quantitative PCR at the indicated time points for GREM1(A), FRZB(B), and DKK1(C). Data expressed as fold-change relative to untreated time point-matched control and represent the mean of three donors ± standard deviation. *P <0.05 compared with unstimulated cells. DKK1, dickkopf 1 homolog (Xenopus laevis); FRZB, frizzled-related protein; GADPH, glyceraldehyde 3-phosphate dehydrogenase; GREM1, Gremlin 1.
Figure 5. Effects of physiological factor-mediated signaling on mRNA expression of GREM1, FRZB and DKK1. Effects of mechanical loading, oxygen level and medium tonicity mediated signaling on mRNA expression of GREM1, FRZB and DKK1. Chondrocytes encapsulated in a hydrogel received no, intermittent or constant cyclical mechanical loading of 0.5 MPa with a frequency of 0.33 Hz and a loading phase of 50% (A). Chondrocytes were exposed to normoxic or hypoxic culture conditions (B), or were cultured in media with different tonicity with or without FK506 (C). All conditions were analyzed for the mRNA expression of GREM1, FRZB and DKK1 by quantitative PCR after 48 hours. Data expressed as fold-change relative to control and represent the mean of three donors ± standard deviation. *P <0.05 compared with unloaded samples or samples containing medium of 280 mOsm. DKK1, dickkopf 1 homolog (Xenopus laevis); FRZB, frizzled-related protein; GADPH, glyceraldehyde 3-phosphate dehydrogenase; GREM1, Gremlin 1.
Figure 6. Preliminary working model of WNT and BMP signaling feedback loop and perturbation by osteoarthritis factors. (A) WNT and BMP signaling reciprocally regulates the transcription of other antagonists. Exposure of WNT agonists leads to activation of WNT signaling [1]. This activation results in the downregulation of WNT antagonists (for example, FRZB and DKK1), leading to less inhibition of WNT signaling [2]. Additionally, BMP antagonists (for example, GREM1) are downregulated, leading to less inhibition of BMP signaling [3]. Stronger BMP signaling results in the upregulation of WNT antagonists [4], establishing a negative feedback mitigating WNT signaling [5]. This feedback loop allows for tight control of both BMP and WNT signaling in articular cartilage contributing to homeostasis. (B) Established factors that influence cartilage homeostasis also perturb this feedback loop. IL-1β, lack of mechanical stimulation and tonicity all decrease the mRNA levels of WNT and BMP antagonists, possibly resulting in a reset of the feedback loop, and contributing to the loss of cartilage homeostasis. BMP, bone morphogenetic protein; DKK1, dickkopf 1 homolog (Xenopus laevis); FRZB, frizzled-related protein; GADPH, glyceraldehyde 3-phosphate dehydrogenase; GREM1, Gremlin 1; WNT, wingless-type MMTV integration site.
Adams,
Integration of signaling pathways regulating chondrocyte differentiation during endochondral bone formation.
2007, Pubmed
Adams,
Integration of signaling pathways regulating chondrocyte differentiation during endochondral bone formation.
2007,
Pubmed
Blanke,
Transplanted chondrocytes inhibit endochondral ossification within cartilage repair tissue.
2009,
Pubmed
Bos,
Increased type II deiodinase protein in OA-affected cartilage and allelic imbalance of OA risk polymorphism rs225014 at DIO2 in human OA joint tissues.
2012,
Pubmed
Chan,
Increased chondrocyte sclerostin may protect against cartilage degradation in osteoarthritis.
2011,
Pubmed
Corallini,
Circulating levels of frizzled-related protein (FRZB) are increased in patients with early rheumatoid arthritis and decrease in response to disease-modifying antirheumatic drugs.
2010,
Pubmed
Daheshia,
The interleukin 1beta pathway in the pathogenesis of osteoarthritis.
2008,
Pubmed
Dreier,
Hypertrophic differentiation of chondrocytes in osteoarthritis: the developmental aspect of degenerative joint disorders.
2010,
Pubmed
Engler,
Substituted 3-imidazo[1,2-a]pyridin-3-yl- 4-(1,2,3,4-tetrahydro-[1,4]diazepino-[6,7,1-hi]indol-7-yl)pyrrole-2,5-diones as highly selective and potent inhibitors of glycogen synthase kinase-3.
2004,
Pubmed
Fischer,
Human articular chondrocytes secrete parathyroid hormone-related protein and inhibit hypertrophy of mesenchymal stem cells in coculture during chondrogenesis.
2010,
Pubmed
Gazzerro,
Conditional deletion of gremlin causes a transient increase in bone formation and bone mass.
2007,
Pubmed
Gelse,
Molecular differentiation between osteophytic and articular cartilage--clues for a transient and permanent chondrocyte phenotype.
2012,
Pubmed
Gómez-Barrena,
Sequential changes of parathyroid hormone related protein (PTHrP) in articular cartilage during progression of inflammatory and degenerative arthritis.
2004,
Pubmed
Haapala,
Remobilization does not fully restore immobilization induced articular cartilage atrophy.
1999,
Pubmed
He,
BMP signaling inhibits intestinal stem cell self-renewal through suppression of Wnt-beta-catenin signaling.
2004,
Pubmed
Honsawek,
Dickkopf-1 (Dkk-1) in plasma and synovial fluid is inversely correlated with radiographic severity of knee osteoarthritis patients.
2010,
Pubmed
Jikko,
Inhibition of chondrocyte terminal differentiation and matrix calcification by soluble factors released by articular chondrocytes.
1999,
Pubmed
Kaler,
Macrophage-derived IL-1beta stimulates Wnt signaling and growth of colon cancer cells: a crosstalk interrupted by vitamin D3.
2009,
Pubmed
Kamekura,
Contribution of runt-related transcription factor 2 to the pathogenesis of osteoarthritis in mice after induction of knee joint instability.
2006,
Pubmed
Kawaguchi,
Regulation of osteoarthritis development by Wnt-beta-catenin signaling through the endochondral ossification process.
2009,
Pubmed
Leijten,
Gremlin 1, frizzled-related protein, and Dkk-1 are key regulators of human articular cartilage homeostasis.
2012,
Pubmed
Leijten,
Hypoxia inhibits hypertrophic differentiation and endochondral ossification in explanted tibiae.
2012,
Pubmed
Lepourcelet,
Small-molecule antagonists of the oncogenic Tcf/beta-catenin protein complex.
2004,
Pubmed
,
Xenbase
Lin,
Modulating hedgehog signaling can attenuate the severity of osteoarthritis.
2009,
Pubmed
Luyten,
Wnt signaling and osteoarthritis.
2009,
Pubmed
Ma,
A Wnt/β-catenin negative feedback loop inhibits interleukin-1-induced matrix metalloproteinase expression in human articular chondrocytes.
2012,
Pubmed
Madhavan,
Biomechanical signals exert sustained attenuation of proinflammatory gene induction in articular chondrocytes.
2006,
Pubmed
Martel-Pelletier,
Cytokines and their role in the pathophysiology of osteoarthritis.
1999,
Pubmed
Miclea,
Apc bridges Wnt/β-catenin and BMP signaling during osteoblast differentiation of KS483 cells.
2011,
Pubmed
Minear,
rBMP represses Wnt signaling and influences skeletal progenitor cell fate specification during bone repair.
2010,
Pubmed
Minina,
BMP and Ihh/PTHrP signaling interact to coordinate chondrocyte proliferation and differentiation.
2001,
Pubmed
Moreira Teixeira,
The effect of platelet lysate supplementation of a dextran-based hydrogel on cartilage formation.
2012,
Pubmed
Moreira Teixeira,
High throughput generated micro-aggregates of chondrocytes stimulate cartilage formation in vitro and in vivo.
2012,
Pubmed
Nakayama,
A novel chordin-like BMP inhibitor, CHL2, expressed preferentially in chondrocytes of developing cartilage and osteoarthritic joint cartilage.
2004,
Pubmed
,
Xenbase
Nilsson,
Gradients in bone morphogenetic protein-related gene expression across the growth plate.
2007,
Pubmed
Oh,
Dkk-1 expression in chondrocytes inhibits experimental osteoarthritic cartilage destruction in mice.
2012,
Pubmed
Pan,
TRPV1 activation is required for hypertonicity-stimulated inflammatory cytokine release in human corneal epithelial cells.
2011,
Pubmed
Radin,
Role of mechanical factors in pathogenesis of primary osteoarthritis.
1972,
Pubmed
Reddi,
Interplay between bone morphogenetic proteins and cognate binding proteins in bone and cartilage development: noggin, chordin and DAN.
2001,
Pubmed
Rountree,
BMP receptor signaling is required for postnatal maintenance of articular cartilage.
2004,
Pubmed
Saito,
Transcriptional regulation of endochondral ossification by HIF-2alpha during skeletal growth and osteoarthritis development.
2010,
Pubmed
Saxon,
Sports participation, sports injuries and osteoarthritis: implications for prevention.
1999,
Pubmed
Sen,
Mechanical loading regulates NFATc1 and beta-catenin signaling through a GSK3beta control node.
2009,
Pubmed
Shanfield,
Synovial fluid osmolality in osteoarthritis and rheumatoid arthritis.
1988,
Pubmed
Spitters,
A dual flow bioreactor with controlled mechanical stimulation for cartilage tissue engineering.
2013,
Pubmed
Stewart,
Elevated expression of hypoxia inducible factor-2alpha in terminally differentiating growth plate chondrocytes.
2006,
Pubmed
Thomas,
Effects of Wnt3A and mechanical load on cartilage chondrocyte homeostasis.
2011,
Pubmed
Tuli,
Transforming growth factor-beta-mediated chondrogenesis of human mesenchymal progenitor cells involves N-cadherin and mitogen-activated protein kinase and Wnt signaling cross-talk.
2003,
Pubmed
van de Loo,
Effects of murine recombinant interleukin 1 on synovial joints in mice: measurement of patellar cartilage metabolism and joint inflammation.
1990,
Pubmed
van der Kraan,
Bone morphogenetic proteins and articular cartilage: To serve and protect or a wolf in sheep clothing's?
2010,
Pubmed
van der Windt,
Physiological tonicity improves human chondrogenic marker expression through nuclear factor of activated T-cells 5 in vitro.
2010,
Pubmed
Wang,
Thyroid hormone interacts with the Wnt/beta-catenin signaling pathway in the terminal differentiation of growth plate chondrocytes.
2007,
Pubmed
Wilson,
The status of Wnt signalling regulates neural and epidermal fates in the chick embryo.
2001,
Pubmed
,
Xenbase
Zhang,
A role for the BMP antagonist chordin in endochondral ossification.
2002,
Pubmed
Zheng,
Type X collagen gene regulation by Runx2 contributes directly to its hypertrophic chondrocyte-specific expression in vivo.
2003,
Pubmed
Zhu,
Inhibition of beta-catenin signaling in articular chondrocytes results in articular cartilage destruction.
2008,
Pubmed
Zhu,
Activation of beta-catenin signaling in articular chondrocytes leads to osteoarthritis-like phenotype in adult beta-catenin conditional activation mice.
2009,
Pubmed
