Identification of key genes and their correlation with immune infiltration in osteoarthritis using integrative bioinformatics approaches and machine-learning strategies.

Osteoarthritis (OA) is a common degenerative joint disease and is closely associated with chronic, low-grade inflammation. Regulating ferroptosis by targeting ferroptosis-related genes may be a fast and effective way to delay the degeneration of OA. However, the molecular mechanisms and gene targets related to ferroptosis in OA are still unclear. Data of OA samples from 3 gene expression omnibus (GEO) datasets were combined to identify differentially expressed genes (DEGs). Ferroptosis-related genes (FRGs) retrieved by the Ferroptosis database were intersected with DEGs, and the intersected hub genes were used for functional enrichment analysis. The feature genes were obtained from the least absolute shrinkage and selection operator (LASSO) algorithm, support vector machine recursive feature elimination (SVM-RFE) algorithm, and random forest (RF) algorithm. Single sample gene set enrichment analysis (ssGSEA) was used to compare immune infiltration between OA patients and normal controls, and the correlation between feature genes and immune cells was analyzed. The expression levels of feature genes were confirmed by RT-PCR. In addition, to explore the applicability of these genes, we extended the bioinformatics analysis of these feature genes to cancer. Finally, 4 feature genes, GABARAPL1, TNFAIP3, ARNTL, and JUN, were confirmed in OA. Theirs expression level were validated by RT-PCR. ROC curves of the 4 genes exhibit excellent diagnostic efficiency for OA, suggesting that the 4 genes were associated with the pathogenesis of OA. Another GEO dataset validated this result. Further analysis revealed that the 4 feature genes were all closely related to the immune infiltration cells in OA. Additionally, results of prognosis analysis indicated that JUN might be a promising therapeutic target for cancer. GABARAPL1, TNFAIP3, ARNTL, and JUN may be predicted biomarkers for OA. The feature genes and association between feature genes and immune infiltration may provide potential biomarkers for OA prediction along with the better assessment of the disease.

Xia D ，Wang J ，Yang S ，Jiang C ，Yao J ... -  《-》

Identification of four-gene signature to diagnose osteoarthritis through bioinformatics and machine learning methods.

Although osteoarthritis (OA) is one of the most prevalent joint disorders, effective biomarkers to diagnose OA are still unavailable. This study aimed to acquire some key synovial biomarkers (hub genes) and analyze their correlation with immune infiltration in OA. Gene expression profiles and clinical characteristics of OA and healthy synovial samples were retrieved from the Gene Expression Omnibus (GEO) database. Hub genes for OA were mined based on a combination of weighted gene co-expression network analysis (WGCNA), the least absolute shrinkage and selection operator (LASSO), support vector machine recursive feature elimination (SVM-RFE), and random forest (RF) algorithms. A diagnostic nomogram model for OA prediction was developed based on the hub genes. Receiver operating characteristic curves (ROC) were performed to confirm the abnormal expression of hub genes in the experimemtal and validation datasets. qRT-PCR using patients' samples were conducted as well. In addition, the infiltration level of 28 immune cells in the expression profile and their relationship with hub genes were analyzed using single-sample GSEA (ssGSEA). 4 hub genes (ZBTB16, TNFSF11, SCRG1 and KDELR3) were obtained by WGCNA, lasso, SVM-RFE, RF algorithms as potential biomarkers for OA. The immune infiltration analyses revealed that hub genes were most correlated with regulatory T cell and natural killer cell. A machine learning model to diagnose OA based on ZBTB16, TNFSF11, SCRG1 and KDELR3 using synovial tissue was constructed, providing theoretical foundation and guideline for diagnostic and treatment targets in OA.

Chen Z ，Wang W ，Zhang Y ，Xue X ，Hua Y ... -  《-》

Identification of ion channel-related genes as diagnostic markers and potential therapeutic targets for osteoarthritis through bioinformatics and machine learning-based approaches.

Yongming L ，Yizhe X ，Zhikai Q ，Yupeng W ，Xiang W ，Mengyuan Y ，Guoqing D ，Hongsheng Z ... -  《-》

Identification of aging-related biomarkers and immune infiltration characteristics in osteoarthritis based on bioinformatics analysis and machine learning.

Osteoarthritis (OA) is a degenerative disease closely related to aging. Nevertheless, the role and mechanisms of aging in osteoarthritis remain unclear. This study aims to identify potential aging-related biomarkers in OA and to explore the role and mechanisms of aging-related genes and the immune microenvironment in OA synovial tissue. Normal and OA synovial gene expression profile microarrays were obtained from the Gene Expression Omnibus (GEO) database and aging-related genes (ARGs) from the Human Aging Genomic Resources database (HAGR). Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), Disease Ontology (DO), and Gene set variation analysis (GSVA) enrichment analysis were used to uncover the underlying mechanisms. To identify Hub ARDEGs with highly correlated OA features (Hub OA-ARDEGs), Weighted Gene Co-expression Network Analysis (WGCNA) and machine learning methods were used. Furthermore, we created diagnostic nomograms and receiver operating characteristic curves (ROC) to assess Hub OA-ARDEGs' ability to diagnose OA and predict which miRNAs and TFs they might act on. The Single sample gene set enrichment analysis (ssGSEA) algorithm was applied to look at the immune infiltration characteristics of OA and their relationship with Hub OA-ARDEGs. We discovered 87 ARDEGs in normal and OA synovium samples. According to functional enrichment, ARDEGs are primarily associated with inflammatory regulation, cellular stress response, cell cycle regulation, and transcriptional regulation. Hub OA-ARDEGs with excellent OA diagnostic ability were identified as MCL1, SIK1, JUND, NFKBIA, and JUN. Wilcox test showed that Hub OA-ARDEGs were all significantly downregulated in OA and were validated in the validation set and by qRT-PCR. Using the ssGSEA algorithm, we discovered that 15 types of immune cell infiltration and six types of immune cell activation were significantly increased in OA synovial samples and well correlated with Hub OA-ARDEGs. Synovial aging may promote the progression of OA by inducing immune inflammation. MCL1, SIK1, JUND, NFKBIA, and JUN can be used as novel diagnostic biomolecular markers and potential therapeutic targets for OA.

Zhou J ，Huang J ，Li Z ，Song Q ，Yang Z ，Wang L ，Meng Q ... -  《Frontiers in Immunology》

Machine learning identifies ferroptosis-related genes as potential diagnostic biomarkers for osteoarthritis.

Osteoarthritis (OA) is one of the most common forms of degenerative arthritis and a major cause of pain and disability. Ferroptosis, a novel mode of cell death, has been verified to participate in the development of OA, but its mechanism is still unclear. This paper analyzed the ferroptosis-related genes (FRGs) in OA and explored their potential clinical value. We downloaded data through the GEO database and screened for DEGs. Subsequently, FRGs were obtained using two machine learning methods, LASSO regression and SVM-RFE. The accuracy of the FRGs as disease diagnosis was identified using ROC curves and externally validated. The CIBERSORT analyzed the immune microenvironment rug regulatory network constructed through the DGIdb. The competitive endogenous RNA (ceRNA) visualization network was constructed to search for possible therapeutic targets. The expression levels of FRGs were verified by qRT-PCR and immunohistochemistry. In this study, we found 4 FRGs. The ROC curve showed that the combined 4 FRGs had the highest diagnostic value. Functional enrichment analysis showed that the 4 FRGs in OA could influence the development of OA through biological oxidative stress, immune response, and other processes. qRT-PCR and immunohistochemistry verified the expression of these key genes, further confirming our findings. Monocytes and macrophages are heavily infiltrated in OA tissues, and the persistent state of immune activation may promote the progression of OA. ETHINYL ESTRADIOL was a possible targeted therapeutic agent for OA. Meanwhile, ceRNA network analysis identified some lncRNAs that could regulate the FRGs. We identify 4 FRGs (AQP8, BRD7, IFNA4, and ARHGEF26-AS1) closely associated with bio-oxidative stress and immune response, which may become early diagnostic and therapeutic targets for OA.

Qiu Y ，Yao J ，Li L ，Xiao M ，Meng J ，Huang X ，Cai Y ，Wen Z ，Huang J ，Zhu M ，Chen S ，Long X ，Li J ... -  《Frontiers in Endocrinology》