PDF(Pubmed)
Abstract：
The etiologies of newborn deaths in neonatal intensive care units usually remain unknown, even after genetic testing. Whole-genome sequencing, combined with artificial intelligence-based methods for predicting the effects of non-coding variants, provide an avenue for resolving these deaths. Using one such method, SpliceAI, we identified a maternally inherited deep intronic PKHD1 splice variant (chr6:52030169T>C), in trans with a pathogenic missense variant (p.Thr36Met), in a newborn who died of autosomal recessive polycystic kidney disease at age 2 days. We validated the deep intronic variant\'s impact in maternal urine-derived cells expressing PKHD1. Reverse transcription polymerase chain reaction followed by Sanger sequencing showed that the variant causes inclusion of 147bp of the canonical intron between exons 29 and 30 of PKHD1 into the mRNA, including a premature stop codon. Allele-specific expression analysis at a heterozygous site in the mother showed that the mutant allele completely suppresses canonical splicing. In an unrelated healthy control, there was no evidence of transcripts including the novel splice junction. We returned a diagnostic report to the parents, who underwent in vitro embryo selection.
摘要：
新生儿重症监护病房新生儿死亡的病因通常仍然未知，甚至在基因检测之后.全基因组测序,结合基于人工智能的方法来预测非编码变体的影响，提供解决这些死亡的途径。使用一种这样的方法，SpliceAI,我们确定了一个母系遗传的深内含子PKHD1剪接变体（chr6：52030169T>C），具有致病性错义变体的反式（p。Thr36Met），一名婴儿在两岁时死于常染色体隐性遗传性多囊肾病。我们验证了深层内含子变异对表达PKHD1的母体尿液来源细胞的影响。逆转录聚合酶链反应，然后进行Sanger测序表明，该变体导致PKHD1外显子29和30之间的经典内含子147bp包含到mRNA中，包括提前终止密码子.母亲杂合位点的等位基因特异性表达分析表明，突变等位基因完全抑制了经典剪接。在一个无关的健康对照中，没有证据表明转录本包括新的剪接接头。我们给父母回了一份诊断报告，接受体外胚胎选择的人。索引词:SpliceAI,内含子,PKHD1,ARPKD,NICU,死者。
