The integrative analysis of competitive endogenous RNA regulatory networks in osteoporosis

Osteoporosis (OP) is a common bone disease of old age resulting from the imbalance between bone resorption and bone formation. CircRNAs are a class of endogenous non-coding RNAs (ncRNAs) involved in gene regulation and may play important roles in the development of OP. Here, we aimed to discover the OP‑related circRNA-miRNA-mRNA (ceRNA) network and the potential mechanisms. Six microarray datasets were obtained from the GEO database and the OP‑related differentially expressed genes (DEGs), circRNAs (DECs), and miRNAs (DEMs) were screened out from these datasets. Then, combined with the prediction of the relationships between DEGs, DEMs, and DECs, a ceRNA network containing 7 target circRNAs, 5 target miRNAs, and 38 target genes was constructed. Then the RNA-seq verification by using total RNAs isolated from the femurs of normal and ovariectomized Wistar rats indicated that MFAP5, CAMK2A, and RGS4 in the ceRNA network were closely associated with osteoporosis. Function enrichment analysis indicated that the target circRNAs, miRNAs, and genes were involved in the process of MAPK cascade, hormone stimulus, cadherin binding, rRNA methyltransferase, PI3K-Akt signaling pathway, and Vitamin digestion and absorption, etc. Then a circRNA-miRNA-hub gene subnetwork was constructed and the qRT-PCR analysis of human bone tissues from the femoral head was used to confirm that the transcription of hsa_circR_0028877, hsa_circR_0082916, DIRAS2, CAMK2A, and MAPK4 showed a significant correlation with osteogenic genes. Besides, the two axes of hsa_circR_0028877/hsa-miR-1273f/CAMK2A and hsa_circR_0028877/hsa-miR-1273f/DIRAS2 conformed to be closely associated with OP. Additionally, by constructing a drug-target gene network, RKI-1447, FRAX486, Hyaluronic, and Fostamatinib were identified as therapeutic options for OP. Our study revealed the potential links between circRNAs, miRNAs, and mRNAs in OP, suggesting that the ceRNA mechanism might contribute to the occurrence of OP.