RNA splicing analysis contributes to reclassifying variants of uncertain significance and improves the diagnosis of monogenic disorders. Issue 10 (4th February 2022)

• Record Type: Journal Article
• Authors: He, Wen-Bin; Xiao, Wen-Juan; Dai, Cong-Ling; Wang, Yu-Rong; Li, Xiu-Rong; Gong, Fei; Meng, Lan-Lan; Tan, Chen; Zeng, Si-Cong; Lu, Guang-Xiu; Lin, Ge; Tan, Yue-Qiu; Hu, Hao; Du, Juan
• Is Part Of: Journal of medical genetics. Volume 59:Issue 10(2022)
• Page Start: 1010
• Page End: 1016
• Publication Date: 2022-02-04
• Subjects: genetic testing -- mutation -- sequence analysis -- RNA; Medical genetics -- Periodicals; 616.042
• DOI: 10.1136/jmedgenet-2021-108013
• Languages: English
• ISSNs: 1468-6244

Abstract

Abstract : Background: Numerous variants of uncertain significance (VUSs) have been identified by whole exome sequencing in clinical practice. However, VUSs are not currently considered medically actionable. Objective: To assess the splicing patterns of 49 VUSs in 48 families identified clinically to improve genetic counselling and family planning. Methods: Forty-nine participants with 49 VUSs were recruited from the Reproductive and Genetic Hospital of CITIC-Xiangya. Bioinformatic analysis was performed to preliminarily predict the splicing effects of these VUSs. RT-PCR and minigene analysis were used to assess the splicing patterns of the VUSs. According to the results obtained, couples opted for different methods of reproductive interventions to conceive a child, including prenatal diagnosis and preimplantation genetic testing (PGT). Results: Eleven variants were found to alter pre-mRNA splicing and one variant caused nonsense-mediated mRNA decay, which resulted in the reclassification of these VUSs as likely pathogenic. One couple chose to undergo in vitro fertilisation with PGT treatment; a healthy embryo was transferred and the pregnancy is ongoing. Three couples opted for natural pregnancy with prenatal diagnosis. One couple terminated the pregnancy because the fetus was affected by short-rib thoracic dysplasia and harboured the related variant. The infants of the other two couples were born and were healthy at their last recorded follow-up. Conclusion: RNA splicing analysis is an important method to assess the impact of sequence variants on splicing in clinical practice and can contribute to the reclassification of a significant proportion of VUSs. RNA splicing analysis should be considered for genetic disease diagnostics.