Department of Molecular Diagnostics, Institute of Oncology Ljubljana, SI-1000 Ljubljana, Slovenia.
Biotechnical Faculty, University of Ljubljana, SI-1000 Ljubljana, Slovenia.
Int J Mol Sci. 2022 Jul 4;23(13):7446. doi: 10.3390/ijms23137446.
Pathogenic/likely pathogenic variants in susceptibility genes that interrupt RNA splicing are a well-documented mechanism of hereditary cancer syndromes development. However, if RNA studies are not performed, most of the variants beyond the canonical GT-AG splice site are characterized as variants of uncertain significance (VUS). To decrease the VUS burden, we have bioinformatically evaluated all novel VUS detected in 732 consecutive patients tested in the routine genetic counseling process. Twelve VUS that were predicted to cause splicing defects were selected for mRNA analysis. Here, we report a functional characterization of 12 variants located beyond the first two intronic nucleotides using RNAseq in , , , , , , , and genes. Based on the analysis of mRNA, we have successfully reclassified 50% of investigated variants. 25% of variants were downgraded to likely benign, whereas 25% were upgraded to likely pathogenic leading to improved clinical management of the patient and the family members.
致病性/可能致病性变异在易感性基因中，这些基因中断 RNA 剪接是遗传性癌症综合征发展的一个有充分文献记录的机制。然而，如果不进行 RNA 研究，大多数超出规范 GT-AG 剪接位点的变异将被归类为意义不明的变异 (VUS)。为了降低 VUS 负担，我们对在常规遗传咨询过程中测试的 732 名连续患者中检测到的所有新的 VUS 进行了生物信息学评估。选择了 12 个预测会导致剪接缺陷的 VUS 进行 mRNA 分析。在这里，我们使用 RNAseq 报告了位于第一个两个内含子核苷酸之外的 12 个变体的功能特征，、、、、、、、和 基因。基于对 mRNA 的分析，我们成功地重新分类了 50%的研究变体。25%的变体降级为可能良性，而 25%的变体升级为可能致病性，从而改善了对患者和家庭成员的临床管理。
