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Neutrophil extracellular traps drive osteoporosis via NCF2-dependent signaling: integrated transcriptomics with
Xiangyun Guo1,2, Liang Wang1, Shuangliu Chen3
1School of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, People's Republic of China.
Background:
Inflammation and immune responses play key roles in osteoporosis (OP) pathogenesis. Osteoimmunology highlights immune dysregulation as a significant contributor to OP, but the specific biological mechanisms linking immune dysfunction to bone loss remain unclear. Understanding these mechanisms is essential for targeted therapies.
Methods:
We established a rat OP model via bilateral ovariectomy. Transcriptomic sequencing (RNA-seq) identified differentially expressed genes (DEGs), and summary data-based Mendelian randomization (SMR) analysis validated their causal associations with OP. Primary neutrophils isolated from bone marrow and differentiated HL-60 neutrophil-like cells were induced to form neutrophil extracellular traps (NETs), and siRNA was employed to knock down the NCF2 gene. Conditioned media from these neutrophils were subsequently applied to primary osteoblasts to evaluate effects on osteogenic differentiation.
Results:
RNA-seq identified 4,497 DEGs (1,606 upregulated, 2,891 downregulated) in OP rats, significantly enriched in immune response and NETs formation pathways. NETs markers (NE, MPO, CitH3) were markedly elevated in OP bone tissue and stimulated neutrophils. SMR analysis identified VDAC1, PLCG2, and NCF2 as key genes significantly associated with OP risk, experimentally validated at the tissue and cellular levels. Knockdown of NCF2 reduced NETs formation in neutrophil-like cells and alleviated NETs-induced osteoblast differentiation impairment. Drug prediction and molecular docking analyses demonstrated high affinity and pharmacological potential targeting these genes.
Conclusions:
This study unveils the link between NETs formation and OP, highlighting NCF2 as crucial players. These findings provide new insights into immune inflammation's role in bone metabolism and pave the way for targeted OP therapies.
Insights
Neutrophil extracellular traps (NETs) contribute to osteoporosis (OP) by impairing osteoblast function. Targeting the NCF2 gene shows potential for novel OP therapies by reducing NETs formation and protecting bone metabolism.
Area of Science:
- Osteoimmunology
- Molecular Biology
- Bone Metabolism
Background:
- Inflammation and immune responses are critical in osteoporosis (OP) pathogenesis.
- Osteoimmunology research indicates immune dysregulation contributes significantly to OP.
- Specific mechanisms linking immune dysfunction to bone loss require elucidation for targeted therapies.
Purpose of the Study:
- To investigate the role of neutrophil extracellular traps (NETs) in osteoporosis.
- To identify key genes and pathways involved in immune-mediated bone loss.
- To explore potential therapeutic targets for OP.
Main Methods:
- Established a rat osteoporosis model using bilateral ovariectomy.
- Utilized transcriptomic sequencing (RNA-seq) to identify differentially expressed genes (DEGs).
- Performed summary data-based Mendelian randomization (SMR) analysis to validate gene-OP associations.
- Investigated neutrophil extracellular trap (NET) formation and the role of the NCF2 gene in vitro.
- Assessed the impact of NETs on osteoblast differentiation.
Main Results:
- RNA-seq identified 4,497 DEGs in OP rats, enriched in immune response and NETs pathways.
- NETs markers were elevated in OP bone tissue and stimulated neutrophils.
- SMR analysis and experimental validation identified VDAC1, PLCG2, and NCF2 as key genes associated with OP risk.
- NCF2 knockdown reduced NETs formation and mitigated NETs-induced impairment of osteoblast differentiation.
- Drug prediction and molecular docking suggested pharmacological potential for targeting these genes.
Conclusions:
- This study establishes a link between NETs formation and osteoporosis.
- NCF2 is identified as a crucial gene in the NETs-osteoporosis axis.
- Findings offer new insights into immune inflammation's role in bone metabolism and suggest novel therapeutic strategies for OP.
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