Identification of Competing Endogenous RNA Regulatory Networks in Vitamin A Deficiency-Induced Congenital Scoliosis by Transcriptome Sequencing Analysis

Abstract

Background/Aims: Congenital scoliosis (CS) is a result of anomalous development of vertebrae and is frequently associated with somitogenesis malformation. Although noncoding RNAs (ncRNAs) have been recently determined to be involved in the pathogenesis of CS, the competing endogenous RNA (ceRNA) regulatory networks in CS remain largely unknown. Methods: Sequencing was conducted to explore the ncRNA expression profiles in rat embryos (gestation day 9) following vitamin A deficiency (VAD) (n = 9 for the vitamin A deficiency-induced congenital scoliosis (VAD-CS) group and n = 4 for the control group). Real-time reverse transcriptase polymerase chain reaction (RT-PCR) was conducted to verify the expression levels of selected mRNAs, long noncoding RNAs (lncRNAs), circular RNAs (circRNAs), and microRNAs (miRNAs). Bioinformatics analysis was used to discover the possible relationships and functions of the ceRNAs. Results: A total of 749 mRNAs, 56 miRNAs, 685 lncRNAs, and 70 circRNAs were identified to have significantly different expression levels in the two groups. Wnt, PI3K-ATK, FoxO, EGFR, and mTOR were found to be the most significant pathways involved in VAD-CS pathogenesis. The circRNA/miRNA/mRNA and lncRNA/miRNA/mRNA networks of CS were built, and the gene expression mechanisms regulated by ncRNAs were unveiled via the ceRNA regulatory networks. Conclusion: We comprehensively identified ceRNA regulatory networks of embryonic somite development in VAD-CS as well as revealed the contribution of different ncRNA expression profiles. Our data demonstrate the association between mRNAs and ncRNAs in the pathogenic mechanism of CS.

Melatonin protected against the detrimental effects of microRNA‐363 in a rat model of vitamin A‐associated congenital spinal deformities: Involvement of Notch signaling

“…Previous studies suggested that miRNAs play crucial roles in development, including stem cell differentiation, migration, metabolism, apoptosis and invasion . Our previous study also reported that miRNAs were deregulated in the development of VAD‐associated congenital spinal deformities . In this study, we selected one of the differentially expressed miRNAs, namely miR‐363, for further functional characterization.…”

“…Congenital spinal deformity was induced by VAD in rats according to our previous study . Twenty female Wistar rats were randomly divided into the control group (n = 8) and VAD group (n = 12).…”

Circular RNAs in nucleus pulposus cell function and intervertebral disc degeneration

“…An accumulating number of studies have demonstrated that circRNAs play pivotal roles in cellular functions such as proliferation, differentiation, invasion, migration and metabolism, and their deregulation has been documented in different types of diseases, including cancers, neurodegeneration, inflammatory/immune diseases and orthopaedic conditions, such as scoliosis, osteoporosis and osteoarthritis . Recently, it has been reported that circRNAs are involved in the development of IDD .…”

LncRNA‐SULT1C2A regulates Foxo4 in congenital scoliosis by targeting rno‐miR‐466c‐5p through PI3K‐ATK signalling

“…Specifically, lncAIS impacts the function of NF90 and HOXD8 and hinders RUNX2 transcription . Previously, we performed sequencing data analysis to build ceRNA networks in embryonic tissues of rats with VAD‐induced CS and found that, based on Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, a SULT1C2A‐rno‐miR‐466c‐5p‐ Foxo4 axis as well as the Phosphoinositide 3 kinases (PI3K)‐AKT pathway appear to be involved in CS pathogenesis based on bioinformatics prediction and sequence analysis …”

“…18 Previously, we performed sequencing data analysis to build ceRNA networks in embryonic tissues of rats with VAD-induced CS and found that, based on Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, a SULT1C2A-rno-miR-466c-5p-Foxo4 axis as well as the Phosphoinositide 3 kinases (PI3K)-AKT pathway appear to be involved in CS pathogenesis based on bioinformatics prediction and sequence analysis. 19 In the present study, we aimed to confirm the expression of the lncRNA SULT1C2A in a rat model of VAD-induced CS and to explore the molecular mechanism of the SULT1C2A-rno-miR-466c-5p-Foxo4 axis in CS. This study of the effect of ceRNA dysregulation within the pathogenesis of VAD-CS provides insight into the mechanisms of somitogenesis and suggests that SULT1C2A may be a potential target for treating CS.…”