The Inorganic Pyrophosphate Transporter ANK Preserves the Differentiated Phenotype of Articular Chondrocyte

Cailotto, F.; Sébillaud, Sylvie; Netter, Patrick; Jouzeau, Jean‐Yves; Bianchi, Arnaud

doi:10.1074/jbc.m109.050534

Cited by 26 publications

(30 citation statements)

References 55 publications

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“…Therefore, it is possible that extracellular PPi resulting from ANK transport directly or via its hydrolysis to extracellular Pi may affect Wnt/β-catenin signaling in osteogenic precursor cells. This possibility is supported by previous findings showing that extracellular PPi affects Wnt ligand expression and Wnt/β-catenin signaling in articular chondrocytes [44]. …”

Section: Discussionsupporting

confidence: 84%

The role of the progressive ankylosis protein (ANK) in adipogenic/osteogenic fate decision of precursor cells

Minashima

Quirno

Lee

et al. 2017

Bone

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The progressive ankylosis protein (ANK) is a transmembrane protein that transports intracellular pyrophosphate (PPi) to the extracellular milieu. In this study we show increased fatty degeneration of the bone marrow of adult ank/ank mice, which lack a functional ANK protein. In addition, isolated bone marrow stromal cells (BMSCs) isolated from ank/ank mice showed a decreased proliferation rate and osteogenic differentiation potential, and an increased adipogenic differentiation potential compared to BMSCs isolated from wild type (WT) littermates. Wnt signaling pathway PCR array analysis revealed that Wnt ligands, Wnt receptors and Wnt signaling proteins that stimulate osteoblast differentiation were expressed at markedly lower levels in ank/ank BMSCs than in WT BMSCs. Lack of ANK function also resulted in impaired bone fracture healing, as indicated by a smaller callus formed and delayed bone formation in the callus site. Whereas 5 weeks after fracture, the fractured bone in WT mice was further remodeled and restored to original shape, the fractured bone in ank/ank mice was not fully restored and remodeled to original shape. In conclusion, our study provides evidence that ANK plays a critical role in the adipogenic/osteogenic fate decision of adult mesenchymal precursor cells. ANK functions in precursor cells are required for osteogenic differentiation of these cells during adult bone homeostasis and repair, whereas lack of ANK functions favors adipogenic differentiation.

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Section: Discussionsupporting

confidence: 84%

The role of the progressive ankylosis protein (ANK) in adipogenic/osteogenic fate decision of precursor cells

Minashima

Quirno

Lee

et al. 2017

Bone

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“…In rat NP cells, hypoxia regulates Ank gene expression [12] and ANKH expression is increased in degenerated human IVDs [12], leading to the hypothesis that loss of homeostatic regulation of PPi is involved in degeneration and calcification in the IVD. In another study, IL-1β-treated chondrocytes in vitro resulted in a loss of differentiated chondrocyte phenotype (diminished Sox2, aggrecan and collagen II transcripts) and diminished Ank expression [16]. We noted a similar effect upon bovine NP cells treated with IL-1β with or without FasL in which reduced Ank gene expression occurred and this down-regulated Ank expression was attenuated by NCCM.…”

Section: Discussionsupporting

confidence: 57%

Notochordal cells protect nucleus pulposus cells from degradation and apoptosis: implications for the mechanisms of intervertebral disc degeneration

et al. 2011

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IntroductionThe relative resistance of non-chondrodystrophic (NCD) canines to degenerative disc disease (DDD) may be due to a combination of anabolic and anti-catabolic factors secreted by notochordal cells within the intervertebral disc (IVD) nucleus pulposus (NP). Factors known to induce DDD include interleukin-1 beta (IL-1ß) and/or Fas-Ligand (Fas-L). Therefore we evaluated the ability of notochordal cell conditioned medium (NCCM) to protect NP cells from IL-1ß and IL-1ß +FasL-mediated cell death and degeneration.MethodsWe cultured bovine NP cells with IL-1ß or IL-1ß+FasL under hypoxic serum-free conditions (3.5% O2) and treated the cells with either serum-free NCCM or basal medium (Advanced DMEM/F-12). We used flow cytometry to evaluate cell death and real-time (RT-)PCR to determine the gene expression of aggrecan, collagen 2, and link protein, mediators of matrix degradation ADAMTS-4 and MMP3, the matrix protection molecule TIMP1, the cluster of differentiation (CD)44 receptor, the inflammatory cytokine IL-6 and Ank. We then determined the expression of specific apoptotic pathways in bovine NP cells by characterizing the expression of activated caspases-3, -8 and -9 in the presence of IL-1ß+FasL when cultured with NCCM, conditioned medium obtained using bovine NP cells (BCCM), and basal medium all supplemented with 2% FBS.ResultsNCCM inhibits bovine NP cell death and apoptosis via suppression of activated caspase-9 and caspase-3/7. Furthermore, NCCM protects NP cells from the degradative effects of IL-1ß and IL-1ß+Fas-L by up-regulating the expression of anabolic/matrix protective genes (aggrecan, collagen type 2, CD44, link protein and TIMP-1) and down-regulating matrix degrading genes such as MMP-3. Expression of ADAMTS-4, which encodes a protein for aggrecan remodeling, is increased. NCCM also protects against IL-1+FasL-mediated down-regulation of Ank expression. Furthermore, NP cells treated with NCCM in the presence of IL-1ß+Fas-L down-regulate the expression of IL-6 by almost 50%. BCCM does not mediate cell death/apoptosis in target bovine NP cells.ConclusionsNotochordal cell-secreted factors suppress NP cell death by inhibition of activated caspase-9 and -3/7 activity and by up-regulating genes contributing anabolic activity and matrix protection of the IVD NP. Harnessing the restorative powers of the notochordal cell could lead to novel cellular and molecular strategies in the treatment of DDD.

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“…Moreover, eP i was previously shown to enhance the expression of matrix ␥-carboxyglutamic acid (Gla) protein in growth plate chondrocytes, thus inhibiting the mineralization process (33). This suggests that eP i alone could orient the chondrocyte phenotype toward a hypertrophic state, whereas we demonstrated previously that ePP i contributed to the maintenance of the differentiated chondrocyte phenotype (18).…”

Section: Discussioncontrasting

confidence: 48%

“…After 2 h in blocking buffer (TBS, 0.1% Tween (TBST) supplemented with 5% nonfat dry milk), membranes were washed three times with TBST and incubated overnight at 4°C with primary antibodies. The antibodies against ANK and PC-1 (Eurogentec, Angers, France) were used at a 1:500 dilution as we described previously (5,18). Antibodies directed against native and phosphorylated ERK1/2 and PKC␣ (Cell Signaling Technology) as well as antibodies against native (Santa Cruz Biotechnology) and phosphorylated ELK-1 (Ets-like protein-1) (Millipore) were used at 1:500.…”

Section: Methodsmentioning

confidence: 99%

Calcium Input Potentiates the Transforming Growth Factor (TGF)-β1-dependent Signaling to Promote the Export of Inorganic Pyrophosphate by Articular Chondrocyte

Cailotto

Reboul

Sébillaud

et al. 2011

Journal of Biological Chemistry

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Transforming growth factor (TGF)-␤1 stimulates extracellular PP i (ePP i ) generation and promotes chondrocalcinosis, which also occurs secondary to hyperparathyroidism-induced hypercalcemia. We previously demonstrated that ANK was upregulated by TGF-␤1 activation of ERK1/2 and Ca 2؉ -dependent protein kinase C (PKC␣). Thus, we investigated mechanisms by which calcium could affect ePP i metabolism, especially its main regulating proteins ANK and PC-1 (plasma cell membrane glycoprotein-1). We stimulated articular chondrocytes with TGF-␤1 under extracellular (eCa 2؉ ) or cytosolic Ca 2؉ (cCa 2؉ ) modulations. We studied ANK, PC-1 expression (quantitative RT-PCR, Western blotting), ePP i levels (radiometric assay), and cCa 2؉ input (fluorescent probe). Voltage-operated Ca 2؉ -channels (VOC) and signaling pathways involved were investigated with selective inhibitors. Finally, Ank promoter activity was evaluated (gene reporter). TGF-␤1 elevated cCa 2؉ and ePP i levels (by up-regulating Ank and PC-1 mRNA/proteins) in an eCa 2؉ dose-dependent manner. TGF-␤1 effects were suppressed by cCa 2؉ chelation or L-and T-VOC blockade while being mostly reproduced by ionomycin. In the same experimental conditions, the activation of Ras, the phosphorylation of ERK1/2 and PKC␣, and the stimulation of Ank promoter activity were affected similarly. Activation of SP1 (specific protein 1) and ELK-1 (Ets-like protein-1) transcription factors supported the regulatory role of Ca 2؉ . SP1 or ELK-1 overexpression or blockade experiments demonstrated a major contribution of ELK-1, which acted synergistically with SP1 to activate Ank promoter in response to TGF-␤1. TGF-␤1 promotes input of eCa 2؉ through opening of L-and T-VOCs, to potentiate ERK1/2 and PKC␣ signaling cascades, resulting in an enhanced activation of Ank promoter and ePP i production in chondrocyte.The balance between extracellular inorganic pyrophosphate (ePP i ) 3 and extracellular inorganic phosphate (eP i ) is critical for homeostasis of articular cartilage. Indeed, an increase in ePP i supports calcium pyrophosphate dihydrate crystal (CPPD) depositions responsible for articular chondrocalcinosis (ACC) (1). Production of ePP i by articular chondrocytes is dependent on PC-1 (plasma cell membrane glycoprotein-1; also known as ENPP1), an ectoenzyme that produces ePP i from nucleotide triphosphates (2), and on the transporter ANK, which exports PP i across the cell membrane (3). Although ePP i level can be controlled by tissue-nonspecific alkaline phosphatase, which hydrolyzes ePP i into eP i , no expression of this enzyme was detected in healthy articular cartilage compared with osteoarthritic cartilage (4). Therefore, CPPD formation only depends on the expression level of PC-1 and ANK in mature articular cartilage. Transforming growth factor (TGF)-␤1 is a well known inducer of ePP i production by articular chondrocytes (1). We demonstrated previously that ANK was the major contributor of this TGF-␤1-induced ePP i production; meanwhile, PC-1 played a minor role (5). Furthermo...

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The Inorganic Pyrophosphate Transporter ANK Preserves the Differentiated Phenotype of Articular Chondrocyte

Cited by 26 publications

References 55 publications

The role of the progressive ankylosis protein (ANK) in adipogenic/osteogenic fate decision of precursor cells

The role of the progressive ankylosis protein (ANK) in adipogenic/osteogenic fate decision of precursor cells

Notochordal cells protect nucleus pulposus cells from degradation and apoptosis: implications for the mechanisms of intervertebral disc degeneration

Calcium Input Potentiates the Transforming Growth Factor (TGF)-β1-dependent Signaling to Promote the Export of Inorganic Pyrophosphate by Articular Chondrocyte

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