Center for Theragnosis, Biomedical Research Division, Korea Institute of Science and Technology, Seoul, Korea.
Proteomics Core Facility, Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea.
PLoS One. 2022 Jul 7;17(7):e0270884. doi: 10.1371/journal.pone.0270884. eCollection 2022.
To identify potential biomarkers in the plasma that could predict histologic chorioamnionitis (HCA) in women with preterm premature rupture of membranes (PPROM), using shotgun and targeted proteomic analyses.
This retrospective cohort study included 78 singleton pregnant women with PPROM (24-34 gestational weeks) who delivered within 96 h of blood sampling. Maternal plasma samples were analyzed by label-free liquid chromatography-tandem mass spectrometry for proteome profiling in a nested case-control study design (HCA cases vs. non-HCA controls [n = 9 each]). Differential expression of 12 candidate proteins was assessed by multiple reaction monitoring-mass spectrometry (MRM-MS) analysis in individual plasma samples from cases and controls matched by gestational age at sampling (n = 40, cohort 1). A validation study was further performed in an independent study group (n = 38, cohort 2) using ELISA and turbidimetric immunoassay for three differentially expressed proteins.
Shotgun proteomics analyses yielded 18 proteins that were differentially expressed (P < 0.05) between HCA cases and non-HCA controls. MRM-MS analysis of 12 differentially expressed proteins further revealed that the CRP, C4A, and SAA4 levels were significantly increased in women with HCA. A multi-marker panel comprising plasma SAA4 and C4A showed enhanced potential for differentiating HCA from non-HCA women (area under the curve = 0.899). Additional validation of these findings by ELISA assays revealed that the CRP levels were significantly higher in women with HCA than in those without HCA, whereas the plasma levels of C4A and SAA4 did not significantly differ between the two groups.
Plasma C4A, SAA4, and CRP were identified as potential biomarkers for detecting HCA in women with PPROM, based on targeted and shotgun proteomic analyses, showing good accuracy when used as a combined dual-biomarker panel (C4A and SAA4). Nevertheless, ELISA validation of these proteins, except for CRP, may not yield clinically useful markers for predicting HCA.
为了鉴定可能预测早产胎膜早破（PPROM）孕妇发生绒毛膜羊膜炎（HCA）的血浆潜在生物标志物，本研究采用无标记液相色谱串联质谱技术对蛋白质组进行了分析。
本回顾性队列研究纳入了 78 例 24-34 孕周内发生 PPROM 并在采血后 96 小时内分娩的单胎孕妇。采用无标记液相色谱串联质谱技术对这些孕妇的血浆样本进行蛋白质组分析，并采用巢式病例对照研究设计对其进行分析（HCA 病例与非 HCA 对照组[n=9 例]）。通过对匹配了采血时孕周的病例和对照组（n=40 例，队列 1）的单个血浆样本进行多重反应监测-质谱（MRM-MS）分析，评估 12 种候选蛋白的差异表达情况。进一步在另一独立研究组（n=38，队列 2）中使用 ELISA 和比浊免疫分析法对三种差异表达蛋白进行验证。
通过无标记蛋白质组学分析发现，HCA 病例与非 HCA 对照组之间有 18 种蛋白存在差异表达（P＜0.05）。MRM-MS 分析进一步表明，HCA 患者的 CRP、C4A 和 SAA4 水平显著升高。由 SAA4 和 C4A 组成的多标志物组合在区分 HCA 患者与非 HCA 患者方面显示出更强的潜力（曲线下面积=0.899）。ELISA 检测结果进一步验证了这一发现，结果表明，HCA 患者的 CRP 水平显著高于非 HCA 患者，而 C4A 和 SAA4 的血浆水平在两组间无显著差异。
基于靶向和无标记蛋白质组学分析，本研究发现 CRP、C4A、SAA4 可作为预测 PPROM 孕妇发生 HCA 的潜在生物标志物，且当作为联合双标志物组合（C4A 和 SAA4）使用时具有良好的准确性。然而，CRP 以外的这些蛋白的 ELISA 验证可能无法为预测 HCA 提供有临床意义的标志物。
