Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation
Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation
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DOI：
10.1186/s13075-018-1573-4
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发表时间：
2018-05-02
影响因子：
4.9
通讯作者：
Wright, Karina T.
中科院分区：
文献类型：
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作者：
Hulme, Charlotte H.;Wilson, Emma L.;Wright, Karina T.
背景：自体软骨细胞植入（ACI）的失败率约为20%，但其原因尚不完全清楚。需要生物标记物来在手术前预测哪些患者可能会失败，以便提供替代或个性化治疗。我们先前使用无标记定量（LF）与动态范围压缩蛋白质组学方法来评估ACI应答者和非应答者的滑液（SF）。然而，我们能够确定只有少数差异丰富的蛋白质在基线。在本研究中，我们建立在这些先前的研究结果，通过评估该SF中的高丰度蛋白质，提供更全面的蛋白质组学分析，在此基础上，可以了解更多ACI成功或失败的生物学基础。在手术的两个阶段（软骨收获和软骨细胞植入），观察了无应答者（平均Lysholm改善33; n = 14）和无应答者（平均Lysholm降低14; n = 13）。结果：iTRAQ蛋白质组学分析证实了我们之前的发现，即软骨收获后蛋白质组发生了显著的变化（70和54个蛋白质组在应答者和无应答者中分别表现出≥ 2.0倍的变化，且在I期和II期之间p < 0.05）。此外，它突出了ACI应答者和非应答者之间差异丰富的28种蛋白质，这些蛋白质在LF研究中没有发现，其中16种在基线时发生了改变。两种蛋白质（补体C1 s亚组分和基质金属蛋白酶3）的差异表达的生物化学证实。iTRAQ和LF蛋白质组数据集的组合生成了深入的SF蛋白质组信息，用于生成代表ACI成功或失败的相互作用组网络。ACI无应答者中失调的功能通路被确定，包括急性期应答信号转导。结论：确定了几种用于基线预测ACI结果的候选生物标志物。已对ACI应答者和非应答者的SF蛋白质组进行了全面概述，从而更好地了解临床结局的生物学途径，特别是非应答者对软骨收获的差异反应。
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.