AUTHOR=Shi Xin , Li Panpan , Wu Xiang , Shu Jun 

TITLE=Whole-transcriptome sequencing identifies key differentially expressed circRNAs/lncRNAs/miRNAs/mRNAs and linked ceRNA networks in adult degenerative scoliosis

JOURNAL=Frontiers in Molecular Neuroscience

VOLUME=Volume 16 - 2023

YEAR=2023

URL=https://www.frontiersin.org/journals/molecular-neuroscience/articles/10.3389/fnmol.2023.1038816

DOI=10.3389/fnmol.2023.1038816

ISSN=1662-5099

ABSTRACT=Background
With an increasing elderly population, adult degenerative scoliosis (ADS), is forecast to be a prevalent disabling condition in an aging society. Exploring the diversities between common disc degeneration and ADS would contribute to unraveling the etiological mechanisms of ADS. Therefore, we aimed to integrate the circRNA, lncRNA, miRNA, and mRNA expression profiles from normal adults (Normal), patients with disc herniation (LDH), and ADS by whole transcriptome sequencing, which identifies key functional ncRNA and ceRNA networks.
Methods
The fresh whole blood samples (n=3/group) were collected from ADS patients (asymmetric disc degeneration, ADS group), LDH patients (symmetric disc degeneration, LDH group), and healthy volunteers (non-degenerative changes, Normal group), which were examined for mRNA, miRNA, lncRNA, and circRNA expression and screened for differentially expressed (DE) RNAs. Then, Gene Ontology (GO) and KEGG analyses were performed for gene annotation and enrichment pathways on the DE RNAs, which were constructed as a lncRNA-miRNA-mRNA network. Eventually, DE RNAs were validated by qRT-PCR targeting disc nucleus pulposus tissue in ADS and LDH group (n=10 / group).
Results
Compared to the LDH group, we identified 3322 DE mRNAs, 221 DE lncRNAs, 20 DE miRNAs, and 15 DE circRNAs in the ADS. In contrast to Normal, 21 miRNAs and 19 circRNAs were differentially expressed in the ADS. The expression of multiple differentially expressed ncRNAs was confirmed by qRT-PCR analysis to be consistent with the sequencing results. In addition, GO and KEGG analysis demonstrated that most DE mRNAs and ncRNAs target genes are involved in various biological processes, including Endocytosis, Apoptosis, Rap1 signaling pathway, Notch signaling pathway, and others. The constructed lncRNA-miRNA-mRNA co-expression network was primarily related to angiogenesis and regulation. 
Conclusion
By focusing on comparing asymmetric and symmetric disc degeneration, whole-transcriptome sequencing systematically screened for key ncRNAs in the development of ADS, which provided an abundance of valuable candidates for the elucidation of regulatory mechanisms. The DE ncRNAs and the lncRNA-miRNA-mRNA network are intrinsically involved in the regulation of mediator and angiogenesis, which may contribute to the insight into the pathogenesis of ADS.