背景:可变剪接(AS)是遗传调控的主要模式,也是产生结构和功能上不同的mRNA和蛋白质变体的最广泛使用的机制之一。AS的失调可能导致异常的转录和蛋白质产物,导致人类疾病的出现。尽管被认为对调节基因表达很重要,全基因组AS失调,潜在机制,与膝骨关节炎(OA)的临床相关性仍未阐明。因此,在这项研究中,我们阐明并验证了OA进展过程中的AS事件及其调控机制.
结果:在这项研究中,我们确定了人类OA和健康半月板样本之间的差异表达基因。其中,OA相关基因主要富集在细胞外基质组织和骨化等生物学途径中.发现主要的OA相关的调节AS(RAS)事件与OA发展过程中的细胞凋亡有关。凋亡相关基因BCL2L13,XAF1和NF2的表达在OA和健康半月板样品之间显着不同。RNA结合蛋白(RBP)和RAS基因的共变网络的构建表明,差异表达的RBP基因LAMA2和CUL4B可能会调节凋亡基因XAF1和BCL2L13在OA进展过程中发生AS事件。最后,RT-qPCR显示,OA半月板样品中的CUL4B表达显着高于正常对照,并且对照和OA样品中XAF1的AS比率显着不同;这些发现与其预期的表达和调控关系一致。
结论:差异表达的RBPs可能在膝关节OA进展过程中调控凋亡基因的AS。XAF1及其调节器,CUL4B,可能作为这种疾病的新型生物标志物和潜在治疗靶点。
BACKGROUND: Alternative splicing (AS) is a principal mode of genetic regulation and one of the most widely used mechanisms to generate structurally and functionally distinct mRNA and protein variants. Dysregulation of AS may result in aberrant transcription and protein products, leading to the emergence of human diseases. Although considered important for regulating gene expression, genome-wide AS dysregulation, underlying mechanisms, and clinical relevance in knee osteoarthritis (OA) remain unelucidated. Therefore, in this study, we elucidated and validated AS events and their regulatory mechanisms during OA progression.
RESULTS: In this study, we identified differentially expressed genes between human OA and healthy meniscus samples. Among them, the OA-associated genes were primarily enriched in biological pathways such as extracellular matrix organization and ossification. The predominant OA-associated regulated AS (RAS) events were found to be involved in apoptosis during OA development. The expression of the apoptosis-related gene BCL2L13, XAF1, and NF2 were significantly different between OA and healthy meniscus samples. The construction of a
covariation network of RNA-binding proteins (RBPs) and RAS genes revealed that differentially expressed RBP genes LAMA2 and CUL4B may regulate the apoptotic genes XAF1 and BCL2L13 to undergo AS events during OA progression. Finally, RT-qPCR revealed that CUL4B expression was significantly higher in OA meniscus samples than in normal controls and that the AS ratio of XAF1 was significantly different between control and OA samples; these findings were consistent with their expected expression and regulatory relationships.
CONCLUSIONS: Differentially expressed RBPs may regulate the AS of apoptotic genes during knee OA progression. XAF1 and its regulator, CUL4B, may serve as novel biomarkers and potential therapeutic targets for this disease.