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Comprehensive analysis of arachidonic acid metabolism-related genes in diagnosis and synovial immune in osteoarthritis: based on bulk and single-cell RNA sequencing data

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Abstract

Background

Osteoarthritis (OA) is one of degenerative-related arthritis, which can be aggravated by low-grade synovitis. It is known that arachidonic acid (AA) dysmetabolism brings OA synovitis. However, the impact of synovial AA metabolism pathway (AMP) related genes on OA remains uncovered.

Methods

Here, we conducted a comprehensive analysis to explore the impact of AA metabolism genes in OA synovium. We obtained transcriptome expression profiles from three raw datasets related to OA synovium (GSE12021, GSE29746, GSE55235) and identified the hub genes of AA metabolism pathways (AMP) in OA synovium. An OA occurrence diagnostic model was constructed and validated based on the identified hub genes. Then, we explored the correlation between hub gene expression and the immune-related module using CIBERSORT and MCP-counter analysis. The unsupervised consensus clustering analysis and weighted correlation network analysis (WGCNA) were utilized to identify robust clusters of identified genes in each cohort. Moreover, the interaction between the hub genes of AMP and immune cells was elucidated through single-cell RNA (scRNA) analysis by scRNA sequencing data from GSE152815.

Results

We found that the expression of AMP-related genes was up-regulated in OA synovium, and seven hub genes (LTC4S, PTGS2, PTGS1, MAPKAPK2, CBR1, PTGDS, and CYP2U1) were identified. The diagnostic model that combined the identified hub genes showed great clinical validity in diagnosing OA (AUC = 0.979). Moreover, significant associations were noticed between the hub genes' expression, immune cell infiltration, and inflammatory cytokine levels. The 30 OA patients were randomized and clustered into three groups using WGCNA analysis based on the hub genes, and diverse immune status was found in different clusters. Of interest, older patients were more likely to be classified into a cluster with higher levels of inflammatory cytokines IL-6 and less infiltration of immune cells. Based on the scRNA-sequencing data, we found that the hub genes had relatively higher expression in macrophages and B cells than other immune cells. Moreover, inflammation-related pathways were significantly enriched in macrophages.

Conclusion

These results suggest that AMP-related genes are closely involved in alterations of OA synovial inflammation. The transcriptional level of hub genes could serve as a potential diagnostic marker for OA.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding authors upon reasonable request.

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Acknowledgements

We thank all the people who offer help for this study. Co-first authors: Bizhi Tu, Run Fang, and Zheng Zhu contributed equally to this paper.

Funding

This study was supported by Grants from Anhui Key Clinical Speciality Construction Project.

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Author notes

  1. Bizhi Tu, Run Fang and Zheng Zhu have contributed equally to this work and should be considered as co-first authors.

Authors and Affiliations

  1. Department of Orthopedics, The Third Affiliated Hospital of Anhui Medical University, The First People’s Hospital of Hefei, 390 Huaihe Road, Hefei, 230061, Anhui, China

    Bizhi Tu, Run Fang, Zheng Zhu, Guang Chen, Cheng Peng & Rende Ning

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  1. Bizhi Tu
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Contributions

Rende Ning conceived the study idea, revised the manuscript, and provided financial support. BT, RF collected the data and wrote the initial draft. ZZ, CP and GC contributed to the data collection and analysis. All authors approved the final draft of the manuscript. All authors are accountable for all aspects of the work in ensuring related questions' accuracy or integrity. Any parts of the work are appropriately investigated and resolved. NR is the guarantor. The corresponding author attests that all listed authors meet authorship criteria and that no others meeting the criteria have been omitted.

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Correspondence to Rende Ning.

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The authors declare that they have no conflicts of interest in this work.

Ethics approval and consent to participate

Ethical approval for our study was granted by The Committee on Medical Ethics of The Third Affiliated Hospital of Anhui Medical University (Reference number 2022 (69)). Since all the data used in the current study was available online, and no individual patient was involved, it could be confirmed we have obtained all the written informed consent.

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Tu, B., Fang, R., Zhu, Z. et al. Comprehensive analysis of arachidonic acid metabolism-related genes in diagnosis and synovial immune in osteoarthritis: based on bulk and single-cell RNA sequencing data. Inflamm. Res. 72, 955–970 (2023). https://doi.org/10.1007/s00011-023-01720-4

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