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Detecting cathepsin activity in human osteoarthritis via activity-based probes
INTRODUCTION: Lysosomal cathepsins have been reported to contribute to Osteoarthritis (OA) pathophysiology due to their increase in pro-inflammatory conditions. Given the causal role of cathepsins in OA, monitoring their specific activity could provide means for assessing OA severity. To this end, w...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4415352/ https://www.ncbi.nlm.nih.gov/pubmed/25889265 http://dx.doi.org/10.1186/s13075-015-0586-5 |
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author | Ben-Aderet, Louisa Merquiol, Emmanuelle Fahham, Duha Kumar, Ashok Reich, Eli Ben-Nun, Yael Kandel, Leonid Haze, Amir Liebergall, Meir Kosińska, Marta K Steinmeyer, Juergen Turk, Boris Blum, Galia Dvir-Ginzberg, Mona |
author_facet | Ben-Aderet, Louisa Merquiol, Emmanuelle Fahham, Duha Kumar, Ashok Reich, Eli Ben-Nun, Yael Kandel, Leonid Haze, Amir Liebergall, Meir Kosińska, Marta K Steinmeyer, Juergen Turk, Boris Blum, Galia Dvir-Ginzberg, Mona |
author_sort | Ben-Aderet, Louisa |
collection | PubMed |
description | INTRODUCTION: Lysosomal cathepsins have been reported to contribute to Osteoarthritis (OA) pathophysiology due to their increase in pro-inflammatory conditions. Given the causal role of cathepsins in OA, monitoring their specific activity could provide means for assessing OA severity. To this end, we herein sought to assess a cathepsin activity-based probe (ABP), GB123, in vitro and in vivo. METHODS: Protein levels and activity of cathepsins B and S were monitored by immunoblot analysis and GB123 labeling in cultured primary chondrocytes and conditioned media, following stimuli with tumor necrosis factor alpha (TNFα) and/or Interleukin 1 beta (IL-1β). Similarly, cathepsin activity was examined in sections of intact cartilage (IC) and degraded cartilage (DC) regions of OA. Finally, synovial fluid (SF) and serum from donors with no signs of diseases, early OA, late OA and rheumatoid arthritis (RA) patients were analyzed with GB123 to detect distinct activity levels of cathepsin B and S. RESULTS: Cathepsin activity in cell lysates, conditioned media explants and DC sections showed enhanced enzymatic activity of cathepsins B and S. Further histological analysis revealed that cathepsin activity was found higher in superficial zones of DC than in IC. Examining serum and SF revealed that cathepsin B is significantly elevated with OA severity in serum and SF, yet levels of cathepsin S are more correlated with synovitis and RA. CONCLUSIONS: Based on our data, cathepsin activity monitored by ABPs correlated well with OA severity and joint inflammation, directing towards a novel etiological target for OA, which possesses significant translational potential in developing means for non-invasive detection of early signs of OA. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13075-015-0586-5) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4415352 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-44153522015-05-01 Detecting cathepsin activity in human osteoarthritis via activity-based probes Ben-Aderet, Louisa Merquiol, Emmanuelle Fahham, Duha Kumar, Ashok Reich, Eli Ben-Nun, Yael Kandel, Leonid Haze, Amir Liebergall, Meir Kosińska, Marta K Steinmeyer, Juergen Turk, Boris Blum, Galia Dvir-Ginzberg, Mona Arthritis Res Ther Research Article INTRODUCTION: Lysosomal cathepsins have been reported to contribute to Osteoarthritis (OA) pathophysiology due to their increase in pro-inflammatory conditions. Given the causal role of cathepsins in OA, monitoring their specific activity could provide means for assessing OA severity. To this end, we herein sought to assess a cathepsin activity-based probe (ABP), GB123, in vitro and in vivo. METHODS: Protein levels and activity of cathepsins B and S were monitored by immunoblot analysis and GB123 labeling in cultured primary chondrocytes and conditioned media, following stimuli with tumor necrosis factor alpha (TNFα) and/or Interleukin 1 beta (IL-1β). Similarly, cathepsin activity was examined in sections of intact cartilage (IC) and degraded cartilage (DC) regions of OA. Finally, synovial fluid (SF) and serum from donors with no signs of diseases, early OA, late OA and rheumatoid arthritis (RA) patients were analyzed with GB123 to detect distinct activity levels of cathepsin B and S. RESULTS: Cathepsin activity in cell lysates, conditioned media explants and DC sections showed enhanced enzymatic activity of cathepsins B and S. Further histological analysis revealed that cathepsin activity was found higher in superficial zones of DC than in IC. Examining serum and SF revealed that cathepsin B is significantly elevated with OA severity in serum and SF, yet levels of cathepsin S are more correlated with synovitis and RA. CONCLUSIONS: Based on our data, cathepsin activity monitored by ABPs correlated well with OA severity and joint inflammation, directing towards a novel etiological target for OA, which possesses significant translational potential in developing means for non-invasive detection of early signs of OA. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13075-015-0586-5) contains supplementary material, which is available to authorized users. BioMed Central 2015-03-20 2015 /pmc/articles/PMC4415352/ /pubmed/25889265 http://dx.doi.org/10.1186/s13075-015-0586-5 Text en © Ben-Aderet et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Ben-Aderet, Louisa Merquiol, Emmanuelle Fahham, Duha Kumar, Ashok Reich, Eli Ben-Nun, Yael Kandel, Leonid Haze, Amir Liebergall, Meir Kosińska, Marta K Steinmeyer, Juergen Turk, Boris Blum, Galia Dvir-Ginzberg, Mona Detecting cathepsin activity in human osteoarthritis via activity-based probes |
title | Detecting cathepsin activity in human osteoarthritis via activity-based probes |
title_full | Detecting cathepsin activity in human osteoarthritis via activity-based probes |
title_fullStr | Detecting cathepsin activity in human osteoarthritis via activity-based probes |
title_full_unstemmed | Detecting cathepsin activity in human osteoarthritis via activity-based probes |
title_short | Detecting cathepsin activity in human osteoarthritis via activity-based probes |
title_sort | detecting cathepsin activity in human osteoarthritis via activity-based probes |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4415352/ https://www.ncbi.nlm.nih.gov/pubmed/25889265 http://dx.doi.org/10.1186/s13075-015-0586-5 |
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