Single-Cell RNA Sequencing Analysis of Key Candidate Genes; Molecular Modeling and Docking-Based Therapeutic Molecules Exploration of Nature Derived Phytoconstituents for Type 2 Diabetes Mellitus.

Type 2 diabetes mellitus (T2DM) is common metabolic disorder in the middle age population, conferring a heavy worldwide burden. Exact underlying common molecular mechanism of T2DM occurrence is unclear. The purpose of this study is to further explore the molecular mechanism of T2DM through integrated bioinformatic analysis. In this investigation, single cell RNA-sequencing data GSE214517 obtained from the Gene Expression Omnibus (GEO) database, was used for investigating the biomarkers and molecular mechanisms of T2DM. Differentially expressed genes (DEGs) were picked out by limma R bioconductor package. Gene Ontology (GO) and REACTOME pathway enrichment analysis, protein-protein interaction (PPI) network analysis, module analysis, miRNA-hub gene regulatory network analysis, TF-hub gene regulatory network analysis, drug-hub gene interaction network analysis, receiver operating characteristic (ROC) curves analysis, homology modeling, molecular docking and ADMET analysis were performed. In total, 957 DEGs, containing 478 up-regulated genes and 479 down-regulated genes, were identified. The DEGs were mainly enriched in protein metabolic process, anatomical structure development, endomembrane system, nucleoplasm, catalytic activity, ion binding, metabolism of carbohydrates and signal transduction. The hub-genes of RPS28, RUVBL1, RPS29, MRPS12, RPS21, KRAS, RPS3A, JUN, RPL9 and GNAQ might be associated with T2DM. The predicted miRNAs (e.g., hsa-mir-325 and hsa-mir-181c-3p), TFs (STAT4 and NUCKS1) and drug molecules (Becaplermin and Phosphorylisopropane) were found to be significantly correlated with T2DM. Molecular docking analysis revealed that Diosgenin binds to KEAP1 with a binding energy of -9.70 and Diosgenin binds to INSIG2 with a binding energy of -9.81, indicating highly stable binding. This study distinguished hub genes and related signaling pathways that can potentially serve as diagnostic indicators and therapeutic biomarkers for T2DM, thereby improving understanding of the molecular mechanisms involved in T2DM.