人颅缝早闭组织的全蛋白质组分析提示炎症信号与破骨细胞激活在人颅骨缝开放中的关联。
Whole-Proteome Analysis of Human Craniosynostotic Tissue Suggests a Link between Inflammatory Signaling and Osteoclast Activation in Human Cranial Suture Patency.
机构信息
Chicago, Ill.; and Los Angeles, Calif.
From the University of Chicago Pritzker School of Medicine; the Computation Institute, Center for Research Informatics, University of Chicago; the Molecular Oncology Laboratory, Department of Orthopedic Surgery, Section of Neurosurgery, and the Laboratory of Craniofacial Development and Biology, Section of Plastic and Reconstructive Surgery, University of Chicago Medicine; and the Division of Plastic and Reconstructive Surgery, University of California, Los Angeles.
出版信息
Plast Reconstr Surg. 2018 Feb;141(2):250e-260e. doi: 10.1097/PRS.0000000000004025.
BACKGROUND
The pathophysiology of nonsyndromic craniosynostosis remains poorly understood. The authors seek to understand the cause of this condition with a specific focus on how osteoclasts may contribute to craniosynostosis. Here, the authors characterize proteins differentially expressed in patent and fused cranial sutures by comparing their respective proteomes.
METHODS
Fused and patent suture samples were obtained from craniosynostotic patients undergoing surgery at a single academic medical center. Extracted protein from samples was interrogated using mass spectrometry. Differential protein expression was determined using maximum likelihood-based G-test with a q-value cutoffs of 0.5 after correction for multiple hypothesis testing. Immunolocalization of lead protein candidates was performed to validate proteomic findings. In addition, quantitative polymerase chain reaction analysis of corresponding gene expression of proteins of interest was performed.
RESULTS
Proteins differentially expressed in patent versus fused sutures included collagen 6A1 (Col6A1), fibromodulin, periostin, aggrecan, adipocyte enhancer-binding protein 1, and osteomodulin (OMD). Maximum likelihood-based G-test suggested that Col6A1, fibromodulin, and adipocyte enhancer-binding protein 1 are highly expressed in patent sutures compared with fused sutures, whereas OMD is up-regulated in fused sutures compared with patent sutures. These results were corroborated by immunohistochemistry. Quantitative polymerase chain reaction data point to an inverse relationship in proteins of interest to RNA transcript levels, in prematurely fused and patent sutures that potentially describes a feedback loop mechanism.
CONCLUSIONS
Proteome analysis validated by immunohistochemistry may provide insight into the mechanism of cranial suture patency and disease from an osteoclast perspective. The authors results suggest a role of inflammatory mediators in nonsyndromic craniosynostosis. Col6A1 may aid in the regulation of suture patency, and OMD may be involved in premature fusion. Additional validation studies are required.
背景
非综合征性颅缝早闭的病理生理学仍知之甚少。作者旨在通过研究破骨细胞如何促成颅缝早闭，来了解这种疾病的病因。为此，作者通过比较各自的蛋白质组，来描绘在开放和融合颅缝中差异表达的蛋白。
方法
从在一家学术医学中心接受手术的颅缝早闭患者中获取融合和开放颅缝的样本。使用质谱法对样本中的提取蛋白进行检测。使用基于最大似然的 G 检验，对经多重假设检验校正后的 q 值截止值为 0.5 的差异蛋白表达进行了检测。对主要候选蛋白进行免疫组化定位，以验证蛋白质组学结果。此外，还对感兴趣的蛋白的相应基因表达进行了定量聚合酶链反应分析。
结果
在开放与融合颅缝中差异表达的蛋白包括胶原 6A1（Col6A1）、纤维调节素、骨膜蛋白、聚集蛋白聚糖、脂肪细胞增强结合蛋白 1 和骨调节蛋白（OMD）。基于最大似然的 G 检验表明，Col6A1、纤维调节素和脂肪细胞增强结合蛋白 1 在开放颅缝中的表达高于融合颅缝，而 OMD 在融合颅缝中的表达高于开放颅缝。免疫组化结果证实了这一结果。定量聚合酶链反应数据表明，在过早融合和开放的颅缝中，感兴趣的蛋白与 RNA 转录水平呈负相关，这可能描述了一种反馈环机制。
结论
通过免疫组化验证的蛋白质组分析可能为从破骨细胞角度理解颅缝开放和疾病的机制提供线索。作者的研究结果表明，炎症介质在非综合征性颅缝早闭中起作用。Col6A1 可能有助于调节颅缝的开放，而 OMD 可能参与了过早融合。需要进一步的验证研究。
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