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

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.

Unraveling the Molecular Landscape of Congenital Pseudoarthrosis of the Tibia: Insights from a Comprehensive Analysis of 162 Probands

“…Recent studies have identified several biological processes, such as skeletal system development, tissue morphogenesis and cell differentiation/proliferation, are associated to CPT [18,19,53],…”

“…This pathway has been reported to be involved in CPT by quantitative proteomics analysis [19]. The knockdown of AKT2…”

“…The copyright holder for this preprint this version posted July 3, 2023. ; https://doi.org/10.1101/2023.06. 28.23292011 doi: medRxiv preprint periosteum tissues are mainly involved in cell matrix assembly, cell adhesion, AKT-PI3K signal pathway activation, and vascular agglutination of CPT patients [19]. These conclusions indicate the intricate biological processes and molecular regulation pathways associated with bone homeostasis potentially contribute to CPT [20][21][22][23] .…”

“…Functional enrichment analysis of those up-regulated genes uncovered 40 significantly enriched biological processes (threshold P=0.01). These enriched biological processes encompassed skeletal system development (logP=-11.96), tissue remodeling including bone resorption, cell matrix assembly (logP=-10.37) [47,48], regulation of cell adhesion (logP=-8.44), regulation of MAPK cascade [49] (logP=-5.22) and autophagy in bone metabolism [50,51], which have been associated with bone formation and bone remodeling [19,[52][53][54] . Additional enriched biological progress included ossification (e.g., bone mineralization and endochondral ossification) [55], myeloid leukocyte differentiation (e.g., osteoclast differentiation and osteoclast development) [56,57], epithelial cell proliferation (e.g., angiogenesis) [58][59][60][61], inorganic ion homeostasis [62,63], regulation of hormone levels [64,65], negative regulation of response to external stimulus [57,66], Vitamin D receptor pathway [67][68][69], cell migration and chemotaxis [70] (Fig.…”

“…1.c and supplementary table 1, threshold P= 0.05/100=5e-4 by Bonferroni correction). Some of these processes have been reported in the DEPs of CPT patients [18,19,53] and also known to play crucial important role in the metabolism of bone homeostasis. The enriched biological processes identified in the transcriptome analysis provide a comprehensive reflection of the etiology of CPT.…”