鉴定和功能分析介导破骨细胞驱动的骨质疏松进展的基因。

Identification and functional analysis of genes mediating osteoclast-driven progression of osteoporosis.

机构信息

The Third Ward of Orthopaedic Department, General Hospital of Ningxia Medical University, Yinchuan, China.

Institute of Osteoarthropathy, Ningxia Key Laboratory of Clinical and Pathogenic Microbiology, Institute of Medical Sciences, General Hospital of Ningxia Medical University, Yinchuan, China.

出版信息

Sci Prog. 2024 Oct-Dec;107(4):368504241300723. doi: 10.1177/00368504241300723.

DOI:10.1177/00368504241300723

PMID:39587887

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11590132/

Abstract

OBJECTIVE

The pathological mechanism of osteoporosis (OP) involves increased bone resorption mediated by osteoclasts and decreased bone formation mediated by osteoblasts, leading to an imbalance in bone homeostasis. Identifying key molecules in osteoclast-mediated OP progression is crucial for the prevention and treatment of OP.

METHODS

Differential expression analysis and weighted gene co-expression network analysis (WGCNA) were performed on the OP patient datasets from the GEO database. The results were intersected with the differential expression results from the osteoclast differentiation dataset to identify key genes. These key genes were then subjected to disease relevance analysis, and consensus clustering was performed on OP patient samples based on their expression profiles. The subgroups were analyzed for differences, followed by GO, KEGG, GSEA, and GSVA analyses, and immune infiltration. Finally, osteoclast differentiation model was constructed. After validating the success of the model using TRAP and F-actin staining, the differential expression of key genes was validated in vitro via Western blot.

RESULTS

CTRL, ARHGEF5, PPAP2C, VSIG2, and PBLD were identified as key genes. These genes exhibited strong disease relevance (AUC > 0.9). Functional enrichment results also indicated their close association with OP and osteoclast differentiation. In vitro differential expression validation showed that during osteoclast differentiation, CTRL was downregulated, while ARHGEF5, PPAP2C, VSIG2, and PBLD were upregulated, with all differences being statistically significant (< 0.05).

DISCUSSION

Currently, there are no studies on the effects of these five genes on osteoclast differentiation. Therefore, it is meaningful to design in vivo and in vitro perturbation experiments to observe the impact of each gene on osteoclast differentiation and OP progression.

CONCLUSION

CTRL, ARHGEF5, PPAP2C, VSIG2, and PBLD show high potential as molecular targets for basic and clinical research in osteoclast-mediated OP.

摘要

目的

骨质疏松症（OP）的病理机制涉及破骨细胞介导的骨吸收增加和成骨细胞介导的骨形成减少，导致骨稳态失衡。鉴定破骨细胞介导的 OP 进展中的关键分子对于 OP 的预防和治疗至关重要。

方法

对 GEO 数据库中 OP 患者数据集进行差异表达分析和加权基因共表达网络分析（WGCNA）。将结果与破骨细胞分化数据集的差异表达结果进行交集，以鉴定关键基因。然后对这些关键基因进行疾病相关性分析，并根据其表达谱对 OP 患者样本进行共识聚类分析。对亚组进行差异分析，然后进行 GO、KEGG、GSEA 和 GSVA 分析以及免疫浸润分析。最后，构建破骨细胞分化模型。使用 TRAP 和 F-actin 染色验证模型成功后，通过 Western blot 在体外验证关键基因的差异表达。

结果

鉴定出 CTRL、ARHGEF5、PPAP2C、VSIG2 和 PBLD 为关键基因。这些基因表现出很强的疾病相关性（AUC > 0.9）。功能富集结果也表明它们与 OP 和破骨细胞分化密切相关。体外差异表达验证显示，在破骨细胞分化过程中，CTRL 下调，而 ARHGEF5、PPAP2C、VSIG2 和 PBLD 上调，所有差异均具有统计学意义（<0.05）。