背景:常染色体显性多囊肾病(ADPKD)是一种常见的单基因多系统疾病,主要由PKD1基因或PKD2基因突变引起。越来越多的证据表明,其中一些变体,被描述为误解,文献或数据库中的同义或无义突变,通过影响前mRNA剪接过程可能是有害的。
结果:本研究旨在结合预测性生物信息学工具和小基因测定确定这些PKD1和PKD2变体对外显子剪接的影响。因此,在19个候选单核苷酸改变中,11个变体分布在PKD1中(c.7866C>A,c.7960A>G,c.7979A>T,c.7987C>T,c.11248C>G,c.11251C>T,c.11257C>G,c.11257C>T,c.11346C>T,和c.11393C>G)和PKD2(c.1480G>T)被鉴定为导致外显子跳跃。
结论:我们证实了PKD1和PKD2基因中的11个变异体通过干扰经典剪接位点的识别或通过破坏外显子剪接增强子并产生外显子剪接沉默来影响正常剪接。考虑到近年来在PKD1和PKD2基因中鉴定的变异数量的增加,这是迄今为止关于ADPKD相关致病基因中外显子变异体的前mRNA剪接的最全面的研究。这些结果强调了在mRNA水平上评估外显子单核苷酸变体在ADPKD中的作用的重要性。
BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is a common monogenic multisystem disease caused primarily by mutations in the PKD1 gene or
PKD2 gene. There is increasing evidence that some of these variants, which are described as missense, synonymous or nonsense mutations in the literature or databases, may be deleterious by affecting the pre-mRNA splicing process.
RESULTS: This study aimed to determine the effect of these PKD1 and
PKD2 variants on exon splicing combined with predictive bioinformatics tools and minigene assay. As a result, among the 19 candidate single nucleotide alterations, 11 variants distributed in PKD1 (c.7866C > A, c.7960A > G, c.7979A > T, c.7987C > T, c.11248C > G, c.11251C > T, c.11257C > G, c.11257C > T, c.11346C > T, and c.11393C > G) and
PKD2 (c.1480G > T) were identified to result in exon skipping.
CONCLUSIONS: We confirmed that 11 variants in the gene of PKD1 and
PKD2 affect normal splicing by interfering the recognition of classical splicing sites or by disrupting exon splicing enhancers and generating exon splicing silencers. This is the most comprehensive study to date on pre-mRNA splicing of exonic variants in ADPKD-associated disease-causing genes in consideration of the increasing number of identified variants in PKD1 and
PKD2 gene in recent years. These results emphasize the significance of assessing the effect of exon single nucleotide variants in ADPKD at the mRNA level.