Shi Xin, Li Panpan, Wu Xiang, Wang Zhihua, Zhao Gang, Shu Jun
The Second Affiliated Hospital of Kunming Medical University, Kunming Medical University, Kunming, China.
Faculty of Medicine and University Hospital of Cologne, University of Cologne, Cologne, Germany.
Front Genet. 2022 Aug 11;13:902943. doi: 10.3389/fgene.2022.902943. eCollection 2022.
Owing to the intensification of the aging process worldwide, the prevalence of adult degenerative scoliosis (ADS) is increasing at an alarming rate. However, genomic research related to the etiology of ADS is rarely reported worldwide. Since long noncoding RNAs (lncRNAs) play a pivotal role in the progression of human diseases, this study aimed to investigate ADS-associated messenger RNAs (mRNAs) and lncRNAs by RNA sequencing (RNA-seq), as well as performed comprehensive bioinformatics analysis based on the lncRNA-mRNA coexpression network and protein-protein interaction (PPI) network. Initially, six whole blood (WB) samples were obtained from three ADS and three nondegenerative lumbar trauma patients who underwent surgical operation for RNA-seq exploration to construct differential mRNA and lncRNA expression profiles. Subsequently, quantitative RT-PCR (qRT-PCR) was performed to validate three randomly selected differentially expressed mRNAs and lncRNAs derived from the nucleus pulposus (NP) tissue of 14 other subjects (seven ADS patients and seven nondegenerative lumbar trauma patients), respectively. A total of 1,651 upregulated and 1,524 downregulated mRNAs and 147 upregulated and 83 downregulated lncRNAs were screened out from the RNA-Seq data, which constructed coexpression networks to investigate their regulatory interactions further. GO gene function prediction revealed that lncRNA-targeted genes might play a vital role in ADS participation in multiple biological processes such as the AMPK signaling pathway, lysosomes, and ubiquitin-mediated proteolysis, as well as cellular metabolic processes. Moreover, the expression levels of three selected lncRNAs and mRNAs were validated by qRT-PCR, respectively, demonstrating that the relative expression levels were consistent with the RNA-seq data. Notably, the dysregulated RNAs, , , , and , were significantly differentially expressed in ADS WB samples and might serve as potentially regulated genes for research in the future. This study provides the first insight into the altered transcriptome profile of long-stranded noncoding RNAs associated with ADS, which paves the way for further exploration of the clinical biomarkers and molecular regulatory mechanisms for this poorly understood degenerative disease. However, the detailed biological mechanisms underlying these candidate lncRNAs in ADS necessitate further elucidation in future studies.
由于全球老龄化进程的加剧，成人退变性脊柱侧凸（ADS）的患病率正以惊人的速度上升。然而，全球范围内关于ADS病因的基因组研究报道很少。由于长链非编码RNA（lncRNAs）在人类疾病进展中起关键作用，本研究旨在通过RNA测序（RNA-seq）研究与ADS相关的信使RNA（mRNAs）和lncRNAs，并基于lncRNA-mRNA共表达网络和蛋白质-蛋白质相互作用（PPI）网络进行全面的生物信息学分析。最初，从3例ADS患者和3例接受手术的非退行性腰椎创伤患者中获取6份全血（WB）样本进行RNA-seq探索，以构建差异mRNA和lncRNA表达谱。随后，进行定量逆转录PCR（qRT-PCR）以分别验证从其他14名受试者（7例ADS患者和7例非退行性腰椎创伤患者）的髓核（NP）组织中随机选择的3种差异表达的mRNA和lncRNA。从RNA-Seq数据中筛选出总共1651个上调和1524个下调的mRNA以及147个上调和83个下调的lncRNA，构建共表达网络以进一步研究它们的调控相互作用。GO基因功能预测显示，lncRNA靶向基因可能在ADS参与多种生物学过程中起重要作用，如AMPK信号通路、溶酶体和泛素介导的蛋白水解以及细胞代谢过程。此外，分别通过qRT-PCR验证了3种选定的lncRNA和mRNA的表达水平，表明相对表达水平与RNA-seq数据一致。值得注意的是，失调的RNA， ， ， ，和 ，在ADS WB样本中差异显著表达，可能作为未来研究的潜在调控基因。本研究首次深入了解了与ADS相关的长链非编码RNA转录组谱的改变，为进一步探索这种了解甚少的退行性疾病的临床生物标志物和分子调控机制铺平了道路。然而，这些候选lncRNA在ADS中的详细生物学机制需要在未来的研究中进一步阐明。
