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Exploring molecular mechanisms of intra-articular changes in osteonecrosis of femoral head using DIA proteomics and
Gang Zhao1,2,3, Yujie Liu1, Yongjun Zheng2
1Department of Orthopedics, the No.4 Medical Centre, Chinese PLA General Hospital, Beijing, 100048, China.
Purpose:
This study is aimed to delve into the crucial proteins associated with hormonal osteonecrosis of the femoral head (ONFH) and its intra-articular lesions through data-independent acquisition (DIA) proteomics and bioinformatics analysis.
Methods:
We randomly selected samples from eligible ONFH patients and collected samples from the necrotic area of the femoral head and load-bearing cartilage. The control group comprised specimens from the same location in patients with femoral neck fractures. With DIA proteomics, we quantitatively and qualitatively tested both groups and analyzed the differentially expressed proteins (DEPs) between groups. Additionally, we enriched the analysis of DEP functions using gene ontology terms and Kyoto Encyclopedia of Genes and Genomes pathways and verified the key proteins in ONFH through Western blot.
Results:
Proteomics experiment uncovered 937 common DEPs (422 upregulated and 515 downregulated) between the two groups. These DEPs mainly participate in biological processes such as hidden attributes, catalytic activity, molecular function regulators, and structural molecule activity, and in pathways such as starch and sucrose metabolism, ECM-receptor interaction, PI3K-Akt signaling, complement and coagulation cascades, IL-17 signaling, phagosome, transcriptional misregulation in cancers, and focal adhesion. Through protein-protein interaction network target gene analysis and Western blot validation, we identified C3, MMP9, APOE, MPO, LCN2, ELANE, HPX, LTF, and THBS1 as key proteins in ONFH.
Conclusions:
With DIA proteomics and bioinformatics analysis, this study reveals the molecular mechanisms of intra-articular lesions in ONFH. A correlation in the necrotic area and load-bearing cartilage of ONFH at ARCO stages IIIB-IV as well as potential key regulatory proteins was identified. These findings will help more deeply understand the pathogenesis of ONFH and may provide important clues for seeking more effective treatment strategies.
Insights
This study identifies key proteins involved in hormonal osteonecrosis of the femoral head (ONFH) and its joint lesions using advanced proteomics. Findings offer insights into ONFH pathogenesis and potential treatment strategies.
Area of Science:
- Biochemistry
- Molecular Biology
- Orthopedics
Background:
- Hormonal osteonecrosis of the femoral head (ONFH) is a debilitating condition affecting joint health.
- Understanding the molecular mechanisms underlying ONFH and associated intra-articular lesions is crucial for effective treatment.
- Previous research has explored various factors, but a comprehensive proteomic analysis of hormonal ONFH is needed.
Purpose of the Study:
- To identify key proteins associated with hormonal osteonecrosis of the femoral head (ONFH) and its intra-articular lesions.
- To elucidate the molecular mechanisms of these lesions using data-independent acquisition (DIA) proteomics and bioinformatics.
- To uncover potential therapeutic targets for ONFH.
Main Methods:
- Employed data-independent acquisition (DIA) proteomics to quantitatively analyze protein expression in femoral head necrotic areas and load-bearing cartilage from ONFH patients and controls.
- Utilized bioinformatics analysis, including gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment, to analyze differentially expressed proteins (DEPs).
- Validated key identified proteins using Western blot.
Main Results:
- Identified 937 common differentially expressed proteins (DEPs) between ONFH and control groups, with 422 upregulated and 515 downregulated.
- DEPs are primarily involved in catalytic activity, molecular function regulation, and pathways such as ECM-receptor interaction, PI3K-Akt signaling, and complement and coagulation cascades.
- Key proteins implicated in ONFH include C3, MMP9, APOE, MPO, LCN2, ELANE, HPX, LTF, and THBS1.
Conclusions:
- DIA proteomics and bioinformatics analysis revealed molecular mechanisms underlying intra-articular lesions in hormonal ONFH.
- Established a correlation between necrotic areas and load-bearing cartilage in advanced ONFH (ARCO stages IIIB-IV).
- Identified potential key regulatory proteins that may inform future therapeutic strategies for ONFH.
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