Sulfated glycosaminoglycans mediate the effects of FGF2 on the osteogenic potential of rat calvarial osteoprogenitor cells

Ling, Ling; Murali, Sadasivam; Dombrowski, Christian; Haupt, Larisa M.; Stein, Gary S.; Wijnen, André J. van; Nurcombe, Victor; Cool, Simon M.

doi:10.1002/jcp.20760

Cited by 56 publications

(59 citation statements)

References 53 publications

(60 reference statements)

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“…The fact that cell surface CS/DS sulfation is a prerequisite for Fgf2 signaling (Ling et al, 2006;Ramachandra et al, 2014) is supported by the finding that there was a reduced Chondroitin-ACI-lyase-digested highly sulfated CHO-K1 and CHO-K1/Ust CS/DS GAGs were fractioned on a Superdex Peptide column. The column was calibrated with cytochrome C to determine void volume (V 0 ) and glycin to determine total column volume (V t ) (n52).…”

Section: Increased Cs/ds 2-o Sulfation In Cho Cells Affects Proliferamentioning

confidence: 94%

Chondroitin/dermatan 2-O sulfotransferase potentiates Fgf2 induced cell migration

Nikolovska

Spillmann

Seidler

2014

Journal of Cell Science

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Fibroblast growth factor 2 (Fgf2) is involved in several biological functions. Fgf2 requires glycosaminoglycans, like chondroitin and dermatan sulfates (hereafter denoted CS/DS) as co-receptors. CS/ DS are linear polysaccharides composed of repeating disaccharide units and , which can be sulfated. Uronyl 2-O-sulfotransferase (Ust) introduces sulfation at the C2 of IdoUA and GlcUA resulting in over-sulfated units. Here, we investigated the role of Ust-mediated CS/DS 2-O sulfation in Fgf2-induced cell migration. We found that CHO-K1 cells overexpressing Ust contain significantly more CS/DS 2-O sulfated units, whereas Ust knockdown abolished CS/DS 2-O sulfation. These structural differences in CS/DS resulted in altered Fgf2 binding and increased phosphorylation of ERK1/2 (also known as MAPK3 and MAPK1, respectively). As a functional consequence of CS/DS 2-O sulfation and altered Fgf2 binding, cell migration and paxillin activation were increased. Inhibition of sulfation, knockdown of Ust and inhibition of FgfR resulted in reduced migration. Similarly, in 3T3 cells Fgf2 treatment increased migration, which was abolished by Ust knockdown. The proteoglycan controlling the CHO migration was syndecan 1. Knockdown of Sdc1 in CHO-K1 cells overexpressing Ust abolished cell migration. We conclude that the presence of distinctly sulfated CS/DS can tune the Fgf2 effect on cell migration.

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Section: Increased Cs/ds 2-o Sulfation In Cho Cells Affects Proliferamentioning

confidence: 94%

Chondroitin/dermatan 2-O sulfotransferase potentiates Fgf2 induced cell migration

Nikolovska

Spillmann

Seidler

2014

Journal of Cell Science

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“…For assays, sodium pyruvate was removed from the medium for MC3T3-E1 and RUNX2-null cells and the fetal calf serum was reduced to 5% for C2C12 cells. Primary rat calvarial osteoprogenitor cells (rOB) were isolated by collagenase/EDTA digestion as described previously (5). The fraction enriched with osteoprogenitors was maintained in the same medium as that of MC3T3-E1 cells except for additional supplementation with 0.25 mg/ml amphoterin B.…”

Section: Methodsmentioning

confidence: 99%

“…Heparin-growth factor interactions control the activities of susceptible ligands and in particular their interactions with cognate receptors, so dictating mesenchymal lineage progression and cell behavior. Heparin (and the less sulfated heparan sulfates) facilitates the binding of FGF2 to FGFRs, so augmenting osteoblast proliferation and differentiation (1)(2)(3)(4)(5). These GAGs also enhance the activity of BMPs, osteoinductive growth factors that have been clinically approved for spinal fusion, in part by protecting them from enzymatic attack or structural antagonists (6,7).…”

mentioning

confidence: 99%

Synergism between Wnt3a and Heparin Enhances Osteogenesis via a Phosphoinositide 3-Kinase/Akt/RUNX2 Pathway

Ling

Dombrowski

Foong

et al. 2010

Journal of Biological Chemistry

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A new strategy has emerged to improve healing of bone defects using exogenous glycosaminoglycans by increasing the effectiveness of bone-anabolic growth factors. Wnt ligands play an important role in bone formation. However, their functional interactions with heparan sulfate/heparin have only been investigated in non-osseous tissues. Our study now shows that the osteogenic activity of Wnt3a is cooperatively stimulated through physical interactions with exogenous heparin. N-Sulfation and to a lesser extent O-sulfation of heparin contribute to the physical binding and optimal co-stimulation of Wnt3a. Wnt3a-heparin signaling synergistically increases osteoblast differentiation with minimal effects on cell proliferation. Thus, heparin selectively reduces the effective dose of Wnt3a needed to elicit osteogenic, but not mitogenic responses. Mechanistically, Wnt3a-heparin signaling strongly activates the phosphoinositide 3-kinase/Akt pathway and requires the bone-related transcription factor RUNX2 to stimulate alkaline phosphatase activity, which parallels canonical ␤-catenin signaling. Collectively, our findings establish the osteo-inductive potential of a heparinmediated Wnt3a-phosphoinositide 3-kinase/Akt-RUNX2 signaling network and suggest that heparan sulfate supplementation may selectively reduce the therapeutic doses of peptide factors required to promote bone formation.

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“…The important role of FGFR signaling in bone formation is highlighted by the finding that gain-of-function mutations in FGFRs results in premature cranial osteogenesis (33,34). FGFR1 was recently shown to be an important transducer of FGF signals in proliferating osteoblasts (35). In contrast, activated FGFR2 was shown to enhance osteoblast differentiation in Apert syndromic craniosynostosis (36 -41).…”

mentioning

confidence: 99%

Fibroblast Growth Factor Receptor 2 Promotes Osteogenic Differentiation in Mesenchymal Cells via ERK1/2 and Protein Kinase C Signaling

Miraoui

Oudina

Petite

et al. 2009

Journal of Biological Chemistry

135

134

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Mesenchymal stem cells (MSCs)3 have the potential to differentiate into different lineages, including osteoblasts, chondroblasts, and adipocytes (1-7). The osteoblast differentiation program of MSCs is characterized by cell recruitment, which is followed by timely expressed genes including Runx2, alkaline phosphatase (ALP), type I collagen (ColA1), and osteocalcin (OC), which is associated with extracellular matrix mineralization (8 -10). The program of MSC osteogenic differentiation can be induced by soluble molecules such as bone morphogenetic proteins (BMPs) or Wnt proteins that activate several signaling pathways to trigger osteoblast differentiation (11-15). Although various downstream signals are known to promote osteogenic differentiation (16 -20), the molecular mechanisms that control the early stages of MSC osteoblast differentiation are not fully elucidated.Fibroblast growth factors (FGFs) play an important major role in the control of cell proliferation, differentiation, and survival in several tissues including bone (21-24). Notably, FGF2 was found to promote cell growth and osteoblast differentiation in bone marrow-derived mesenchymal cells (25,26). Consistent with an important role of FGF signaling in the control of osteoprogenitor cells, deletion of FGF2 in mice results in decreased bone marrow stromal cell osteogenic differentiation and altered bone formation (27). The actions of FGFs are highly dependent on high affinity FGF receptors (FGFRs) (28). FGF binding to FGFRs leads to receptor dimerization and phosphorylation of intrinsic tyrosine residues, which leads to activation of several signal transduction pathways including phospholipase C␥ (PLC␥), mitogen-activated protein kinases (MAPK), and phosphatidylinositol 3-kinase (PI3K) (29,30). In bone, activation of extracellular-related kinase (ERK1/2) MAPK and protein kinase C (PKC) was found to enhance osteoblast gene expression (31, 32). The important role of FGFR signaling in bone formation is highlighted by the finding that gain-of-function mutations in FGFRs results in premature cranial osteogenesis (33, 34). FGFR1 was recently shown to be an important transducer of FGF signals in proliferating osteoblasts (35). In contrast, activated FGFR2 was shown to enhance osteoblast differentiation in Apert syndromic craniosynostosis (36 -41). However, the role of FGFR2 signaling in osteogenic differentiation of mesenchymal stem cells is yet to be elucidated.In the present study, we investigated the specific role of FGFR2 signaling on osteoblast commitment and differentiation

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Sulfated glycosaminoglycans mediate the effects of FGF2 on the osteogenic potential of rat calvarial osteoprogenitor cells

Cited by 56 publications

References 53 publications

Chondroitin/dermatan 2-O sulfotransferase potentiates Fgf2 induced cell migration

Chondroitin/dermatan 2-O sulfotransferase potentiates Fgf2 induced cell migration

Synergism between Wnt3a and Heparin Enhances Osteogenesis via a Phosphoinositide 3-Kinase/Akt/RUNX2 Pathway

Fibroblast Growth Factor Receptor 2 Promotes Osteogenic Differentiation in Mesenchymal Cells via ERK1/2 and Protein Kinase C Signaling

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