Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation
Abstract Background Autologous chondrocyte implantation (ACI) has a failure rate of approximately 20%, but it is yet to be fully understood why. Biomarkers are needed that can pre-operatively predict in which patients it is likely to fail, so that alternative or individualised therapies can be offer...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-05-01
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| Series: | Arthritis Research & Therapy |
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| Online Access: | http://link.springer.com/article/10.1186/s13075-018-1573-4 |
| _version_ | 1818907904782106624 |
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| author | Charlotte H. Hulme Emma L. Wilson Heidi R. Fuller Sally Roberts James B. Richardson Pete Gallacher Mandy J. Peffers Sally L. Shirran Catherine H. Botting Karina T. Wright |
| author_facet | Charlotte H. Hulme Emma L. Wilson Heidi R. Fuller Sally Roberts James B. Richardson Pete Gallacher Mandy J. Peffers Sally L. Shirran Catherine H. Botting Karina T. Wright |
| author_sort | Charlotte H. Hulme |
| collection | DOAJ |
| description | Abstract Background Autologous chondrocyte implantation (ACI) has a failure rate of approximately 20%, but it is yet to be fully understood why. Biomarkers are needed that can pre-operatively predict in which patients it is likely to fail, so that alternative or individualised therapies can be offered. We previously used label-free quantitation (LF) with a dynamic range compression proteomic approach to assess the synovial fluid (SF) of ACI responders and non-responders. However, we were able to identify only a few differentially abundant proteins at baseline. In the present study, we built upon these previous findings by assessing higher-abundance proteins within this SF, providing a more global proteomic analysis on the basis of which more of the biology underlying ACI success or failure can be understood. Methods Isobaric tagging for relative and absolute quantitation (iTRAQ) proteomic analysis was used to assess SF from ACI responders (mean Lysholm improvement of 33; n = 14) and non-responders (mean Lysholm decrease of 14; n = 13) at the two stages of surgery (cartilage harvest and chondrocyte implantation). Differentially abundant proteins in iTRAQ and combined iTRAQ and LF datasets were investigated using pathway and network analyses. Results iTRAQ proteomic analysis confirmed our previous finding that there is a marked proteomic shift in response to cartilage harvest (70 and 54 proteins demonstrating ≥ 2.0-fold change and p < 0.05 between stages I and II in responders and non-responders, respectively). Further, it highlighted 28 proteins that were differentially abundant between responders and non-responders to ACI, which were not found in the LF study, 16 of which were altered at baseline. The differential expression of two proteins (complement C1s subcomponent and matrix metalloproteinase 3) was confirmed biochemically. Combination of the iTRAQ and LF proteomic datasets generated in-depth SF proteome information that was used to generate interactome networks representing ACI success or failure. Functional pathways that are dysregulated in ACI non-responders were identified, including acute-phase response signalling. Conclusions Several candidate biomarkers for baseline prediction of ACI outcome were identified. A holistic overview of the SF proteome in responders and non-responders to ACI has been profiled, providing a better understanding of the biological pathways underlying clinical outcome, particularly the differential response to cartilage harvest in non-responders. |
| first_indexed | 2024-12-19T22:02:32Z |
| format | Article |
| id | doaj.art-42be40ada3ed4d8b9a54fce0c43c0ef2 |
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| issn | 1478-6362 |
| language | English |
| last_indexed | 2024-12-19T22:02:32Z |
| publishDate | 2018-05-01 |
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| spelling | doaj.art-42be40ada3ed4d8b9a54fce0c43c0ef22022-12-21T20:04:07ZengBMCArthritis Research & Therapy1478-63622018-05-0120111710.1186/s13075-018-1573-4Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte ImplantationCharlotte H. Hulme0Emma L. Wilson1Heidi R. Fuller2Sally Roberts3James B. Richardson4Pete Gallacher5Mandy J. Peffers6Sally L. Shirran7Catherine H. Botting8Karina T. Wright9Institute of Science and Technology in Medicine, Keele UniversityRobert Jones and Agnes Hunt Orthopaedic HospitalInstitute of Science and Technology in Medicine, Keele UniversityInstitute of Science and Technology in Medicine, Keele UniversityInstitute of Science and Technology in Medicine, Keele UniversityInstitute of Science and Technology in Medicine, Keele UniversityInstitute of Ageing and Chronic Disease, University of LiverpoolBSRC Mass Spectrometry and Proteomics Facility, University of St AndrewsBSRC Mass Spectrometry and Proteomics Facility, University of St AndrewsInstitute of Science and Technology in Medicine, Keele UniversityAbstract Background Autologous chondrocyte implantation (ACI) has a failure rate of approximately 20%, but it is yet to be fully understood why. Biomarkers are needed that can pre-operatively predict in which patients it is likely to fail, so that alternative or individualised therapies can be offered. We previously used label-free quantitation (LF) with a dynamic range compression proteomic approach to assess the synovial fluid (SF) of ACI responders and non-responders. However, we were able to identify only a few differentially abundant proteins at baseline. In the present study, we built upon these previous findings by assessing higher-abundance proteins within this SF, providing a more global proteomic analysis on the basis of which more of the biology underlying ACI success or failure can be understood. Methods Isobaric tagging for relative and absolute quantitation (iTRAQ) proteomic analysis was used to assess SF from ACI responders (mean Lysholm improvement of 33; n = 14) and non-responders (mean Lysholm decrease of 14; n = 13) at the two stages of surgery (cartilage harvest and chondrocyte implantation). Differentially abundant proteins in iTRAQ and combined iTRAQ and LF datasets were investigated using pathway and network analyses. Results iTRAQ proteomic analysis confirmed our previous finding that there is a marked proteomic shift in response to cartilage harvest (70 and 54 proteins demonstrating ≥ 2.0-fold change and p < 0.05 between stages I and II in responders and non-responders, respectively). Further, it highlighted 28 proteins that were differentially abundant between responders and non-responders to ACI, which were not found in the LF study, 16 of which were altered at baseline. The differential expression of two proteins (complement C1s subcomponent and matrix metalloproteinase 3) was confirmed biochemically. Combination of the iTRAQ and LF proteomic datasets generated in-depth SF proteome information that was used to generate interactome networks representing ACI success or failure. Functional pathways that are dysregulated in ACI non-responders were identified, including acute-phase response signalling. Conclusions Several candidate biomarkers for baseline prediction of ACI outcome were identified. A holistic overview of the SF proteome in responders and non-responders to ACI has been profiled, providing a better understanding of the biological pathways underlying clinical outcome, particularly the differential response to cartilage harvest in non-responders.http://link.springer.com/article/10.1186/s13075-018-1573-4Autologous chondrocyte implantation (ACI)iTRAQ proteomicsLabel-free quantitation proteomicsSynovial fluidCartilage repairComplement C1S subcomponent |
| spellingShingle | Charlotte H. Hulme Emma L. Wilson Heidi R. Fuller Sally Roberts James B. Richardson Pete Gallacher Mandy J. Peffers Sally L. Shirran Catherine H. Botting Karina T. Wright Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation Arthritis Research & Therapy Autologous chondrocyte implantation (ACI) iTRAQ proteomics Label-free quantitation proteomics Synovial fluid Cartilage repair Complement C1S subcomponent |
| title | Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation |
| title_full | Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation |
| title_fullStr | Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation |
| title_full_unstemmed | Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation |
| title_short | Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation |
| title_sort | two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to autologous chondrocyte implantation |
| topic | Autologous chondrocyte implantation (ACI) iTRAQ proteomics Label-free quantitation proteomics Synovial fluid Cartilage repair Complement C1S subcomponent |
| url | http://link.springer.com/article/10.1186/s13075-018-1573-4 |
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