应用骨膜蛋白质组学分析对儿童先天性胫骨假关节发病机制的新认识。

New insights into pathogenesis of congenital pseudarthrosis of tibia in children using periosteum proteomics analysis.

机构信息

Department of Pediatric Orthopaedics, Hunan Children's Hospital, The School of Pediatrics, Heng Yang Medical School, University of South China, Changsha City, Hunan Province, 410007, China.

Medical School, Fuyang Normal University, Fuyang, China.

出版信息

Rapid Commun Mass Spectrom. 2022 Nov 15;36(21):e9374. doi: 10.1002/rcm.9374.

DOI:10.1002/rcm.9374

PMID:35933588

Abstract

RATIONALE

The exact etiology and pathogenesis of congenital pseudarthrosis of tibia (CPT) are not clear. Quantitative proteomics analysis plays a vital role in disease pathology research. Tandem mass tag (TMT)-based proteomics techniques were employed to identify and analyze the differentially expressed proteins (DEP) in the tibia periosteum tissues of CPT patients.

METHODS

The samples were divided into three groups: CPT with NF1 group, CPT without NF1 group (non-NF1-CPT), and control group (patients with open tibial fracture). A fold change ≥1.5 or ≤0.66 and P-value <0.05 were used as the thresholds to screen DEPs. Subsequently, bioinformatics resources such as online tools DAVID and String were used to generate gene ontology (GO) annotation, KEGG pathways enrichment, and protein-protein interaction (PPI) network for these DEPs.

RESULTS

The results show that a total of 347 proteins were differentially expressed in NF1-CPT groups, 212 of which were upregulated and 135 were downregulated. There were more DEPs in non-NF1-CPT groups; we identified 467 DEPs, including 281 upregulated and 186 downregulated. Among them, NF1-CPT groups and non-NF1-CPT groups shared 231 DEPs, and the remaining 230 DEPs showed the same expression trend in the two disease groups, with 117 upregulated and 113 downregulated. In particular, 116 proteins were altered only in NF1-CPT groups (94 were upregulated and 22 were downregulated), whereas 236 proteins were altered only in non-NF1-CPT groups (164 were upregulated and 72 were downregulated). Finally, compared with non-NF1-CPT groups, 47 proteins changed 1.5-fold and P-value < 0.05 in NF1-CPT groups.

CONCLUSIONS

To sum up, we found that common DEPS in periosteum of NF1-CPT and non-NF1-CPT groups are mainly involved in cell matrix assembly, cell adhesion, AKT-PI3K signal pathway activation, and vascular agglutination, which indicate that these are the pathological characteristics of CPT. The osteogenic ability is weak, the osteoclastic ability is strong, the vascular lumen is narrow, the invasive growth and the proliferation of fibroblasts are enhanced in CPT patients.

摘要

背景

先天性假关节胫骨（CPT）的确切病因和发病机制尚不清楚。定量蛋白质组学分析在疾病病理学研究中起着至关重要的作用。本研究采用串联质量标签（TMT）蛋白质组学技术，鉴定和分析 CPT 患者骨膜组织中的差异表达蛋白（DEP）。

方法

将样本分为三组：NF1 合并 CPT 组（NF1-CPT 组）、不合并 NF1 的 CPT 组（非 NF1-CPT 组）和对照组（胫骨开放性骨折患者）。使用 fold change≥1.5 或≤0.66 和 P 值<0.05 作为筛选 DEP 的阈值。随后，使用在线工具 DAVID 和 String 等生物信息学资源对这些 DEP 进行基因本体（GO）注释、KEGG 通路富集和蛋白质-蛋白质相互作用（PPI）网络分析。

结果

结果显示，NF1-CPT 组中有 347 种蛋白表达差异，其中 212 种上调，135 种下调。非 NF1-CPT 组中 DEP 更多，共鉴定出 467 个 DEP，其中 281 个上调，186 个下调。NF1-CPT 组和非 NF1-CPT 组共有 231 个 DEP，两组中其余 230 个 DEP 的表达趋势相同，其中 117 个上调，113 个下调。特别地，116 个蛋白仅在 NF1-CPT 组中发生改变（94 个上调，22 个下调），而 236 个蛋白仅在非 NF1-CPT 组中发生改变（164 个上调，72 个下调）。最后，与非 NF1-CPT 组相比，NF1-CPT 组中有 47 个蛋白的变化倍数为 1.5 倍，P 值<0.05。