Inhibition of nucleotide pyrophosphatase/ phosphodiesterase 1: Implications for developing a calcium pyrophosphate deposition disease modifying drug

Ortal Danino, Shuli Svetitsky, Sarah Kenigsberg, Asaf Levin, Shani Journo, Aviram Gold, Michael Drexler, Nimrod Snir, Ori Elkayam, Bilha Fischer, Uri Arad*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Objectives. Calcium pyrophosphate deposition (CPPD) is associated with osteoarthritis and is the cause of a common inflammatory articular disease. Ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (eNPP1) is the major ecto-pyrophosphatase in chondrocytes and cartilage-derived matrix vesicles (MVs). Thus, eNPP1 is a principle contributor to extracellular pyrophosphate levels and a potential target for interventions aimed at preventing CPPD. Recently, we synthesized and described a novel eNPP1-specific inhibitor, SK4A, and we set out to evaluate whether this inhibitor attenuates nucleotide pyrophosphatase activity in human OA cartilage. Methods. Cartilage tissue, chondrocytes and cartilage-derived MVs were obtained from donors with OA undergoing arthroplasty. The effect of SK4A on cell viability was assayed by the XTT method. eNPP1 expression was evaluated by western blot. Nucleotide pyrophosphatase activity was measured by a colorimetric assay and by HPLC analysis of adenosine triphosphate (ATP) levels. ATP-induced calcium deposition in cultured chondrocytes was visualized and quantified with Alizarin red S staining. Results. OA chondrocytes expressed eNPP1 in early passages, but this expression was subsequently lost upon further passaging. Similarly, significant nucleotide pyrophosphatase activity was only detected in early-passage chondrocytes. The eNPP1 inhibitor, SK4A, was not toxic to chondrocytes and stable in culture medium and human plasma. SK4A effectively inhibited nucleotide pyrophosphatase activity in whole cartilage tissue, in chondrocytes and in cartilage-derived MVs and reduced ATP-induced CPPD. Conclusion. Nucleotide analogues such as SK4A may be developed as potent and specific inhibitors of eNPP1 for the purpose of lowering extracellular pyrophosphate levels in human cartilage with the aim of preventing and treating CPPD disease.

Original languageEnglish
Pages (from-to)1472-1480
Number of pages9
JournalRheumatology
Volume57
Issue number8
DOIs
StatePublished - 1 Aug 2018
Externally publishedYes

Funding

FundersFunder number
Kamin Fund of the Israel Ministry of Trade and Industry56136

    Keywords

    • CPPD
    • Cartilage
    • ENPP1
    • Inhibitor
    • Osteoarthritis

    Fingerprint

    Dive into the research topics of 'Inhibition of nucleotide pyrophosphatase/ phosphodiesterase 1: Implications for developing a calcium pyrophosphate deposition disease modifying drug'. Together they form a unique fingerprint.

    Cite this