Abstract
Background and Aims
The fact that activation of the innate immune system and chronic inflammation are closely involved in the pathogenesis of diabetic Kidney disease (DKD). Recent studies have suggested the inflammatory process plays a crucial role in the progression of DKD. Identifying novel inflammatory molecules closely related to the decline of renal function is of significance in diagnosing and predicting the progression of DKD. The weighted gene co-expression network analysis (WGCNA) algorithm represents a novel systems biology method that provide the approach of association between gene modules and clinical traits to find the genes involvement into the certain phenotypic trait. The goal of this study was to identify hub genes and their roles in DKD from the gene sets associated with the decline of renal function by WGCNA.
Method
The Gene Expression Omnibus (GEO) database and “Nephroseq” website were searched and transcriptome study from DN biopsies with well-established clinical phenotypic data were selected for analysis. Next, we constructed a weighted gene co-expression network and identified modules negatively correlated with eGFR by WGCNA in the data of glomerular tissue. Functional annotations of the genes in modules negatively correlated with eGFR were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Through protein-protein interaction (PPI) analysis and hub gene screening, the hub genes were obtained. Furthermore, we compared the expression level of hub genes between DKD and normal control and drew ROC curves to determine the diagnosis value to DKD of these genes.
Results
The microarray-based expression datasets GSE30528 were screened out for analysis, which included glomeruli tissue of 9 cases of DKD and 13 cases of control. This microarray platform represented the transcriptome profile of 12411 well-characterized genes. Using WGCNA, a total of 19 gene modules were identified. Then module eigengene were analyzed for correlation with clinical traits of age, sex, ethnicity and eGFR and the “MEhoneydew1” module showed negative associated with eGFR (r=-0.58). GO functional annotation showed that these 551 genes in the “MEhoneydew1” module mainly enriched in the T cell activation. KEGG annotation showed mainly enriched in chemokine signaling pathway. Except for C3, top 10 hub genes, CCR5, CXCR4, CCR7, CCL5, CXCL8, CCR2, CCR1, CX3CR1, C3AR1 and C3, are all members of chemokines or chemokine receptors. Furthermore, we compared the expression level of these 9 genes between DKD and control, and found that all of these 9 genes increased in the DKD group, and the differences of 6 genes, CCR5, CCR7, CCL5, CCR2, CCR1, C3AR1, were of statistical significance. Linear correlation analysis showed that the expression of these 6 genes was negatively correlated with eGFR, and the ROC curve showed that the area under the curve could reach 0.812∼1.0.
Conclusion
We identified a panel of 6 hub genes focused on chemokines and chemokine receptors critical for decline of renal function of DKD using WGCNA. These genes may serve as biomarkers for diagnosis/prognosis and as putative novel therapeutic targets for DKD.
Single-Nucleus Transcriptomic Analysis Reveals Important Cell Cross-Talk in Diabetic Kidney Disease
