Abstract：
Polycystic kidney disease (PKD) is common genetic renal disorder. In present study, we performed WES to identify pathogenic variant in nine families including 26 patients with PKD and 19 unaffected members. The eight pathogenic variants were identified in known PKD associated genes including PKD1 (n = 6), PKD2 (n = 1), and OFD1 (n = 1) in eight families. There is one missense, one stopgain, two non-frameshifts, two canonical splicing variants, three frameshift variants and one potential non-canonical splicing variant (NCSV) in 8 families. The six variants were novel variants and not reported in ClinVar database. In addition, the compound heterozygous variants in PKHD1 were identified including one frameshift variants (PKHD1: NM_138694.4, c.9841del, p.S3281Lfs*4) and one non-canonical splicing variant (PKHD1: NM_138694.4, c.6332 + 40A > G) which were defined as deleterious variant by four splicing prediction tools (CADD-splice, SpliceAI, Spliceogen, Squirl). We used the minigene method to validate whether the prioritized potential NSCVs disrupt the typical mRNA splicing process and found abnormally larger PCR production of minigene carrying potential NCSV comparing to wild-type minigene. Sanger sequencing confirmed the 39-bp insertion of intron 38 between exon 38 and exon 39, which results in non-frameshift and 13 amino acid insertions. In conclusion, our study expands the variant spectrum and highlight the important role of non-canonical splicing variant in PKD.
摘要：
多囊肾病（PKD）是常见的遗传性肾脏疾病。在目前的研究中,我们在9个家庭中进行了WES,其中包括26例PKD患者和19例未受影响的成员.在已知的PKD相关基因包括PKD1（n=6）中鉴定了8种致病变异，PKD2(n=1),和OFD1(n=1)在八个家庭中。有一个错觉，一个止痛药，两个非移相者，两个典型剪接变体，8个家族中的三个移码变体和一个潜在的非规范剪接变体（NCSV）。这6种变体是新的变体，在ClinVar数据库中没有报道。此外,鉴定了PKHD1中的复合杂合变体，包括一个移码变体（PKHD1：NM_138694.4，c.9841del，p.S3281Lfs*4）和一个非规范剪接变体（PKHD1：NM_138694.4，c.633240A>G），通过四个剪接预测工具（CADD-剪接，SpliceAI,剪接原,Squirl）。我们使用小基因方法来验证优先的潜在NSCV是否破坏典型的mRNA剪接过程，并发现与野生型小基因相比，携带潜在NCSV的小基因的PCR产量异常大。Sanger测序证实了内含子38在外显子38和外显子39之间的39-bp插入，这导致非移码和13个氨基酸的插入。总之,我们的研究扩展了变异谱,强调了非规范剪接变异体在PKD中的重要作用.
