LI Juan, WEI Qi, LI Jing-ran, ZHANG Ying, TANG Zhi-xia, ZOU Lin-bing, HONG Ming-yun
Objective To investigate the application value of single nucleotide polymorphism (SNP)linkage analysis based on next-generation sequencing (NGS) technology in preimplantation genetic testing (PGT)of families with autosomal dominant polycystic kidney disease (ADPKD)induced by novel mutations. Methods A family with ADPKD induced by novel mutations was selected,and whole exome sequencing and karyotyping were used to determine the pathogenic mutations of the family. To block the inheritance of the disease,blastocyst culture was performed after in vitro fertilization,blastocyst trophoblast cell samples were biopsied,and multiple annealing and looping-based amplification cycles were used for whole genome amplification of the biopsied cells. Sanger sequencing and next-generation sequencing(NGS)-based single nucleotide polymorphism (SNP) haplotyping were used to detect the state of the gene mutations. Copy number variation (CNV) analysis was used for chromosomal aneuploidy screening of the embryos. Prenatal diagnosis in the second trimester of pregnancy was preformed to verify the PGT outcomes. Results A novel c.2098-2A>G mutation in PKD1 gene was found in the proband. A total of 3 blastocysts formed after intracytoplasmic sperm injection were biopsied. One blastocyst was mutation-free and euploid,and the euploid embryo underwent frozen embryo transplantation (FET) to achieve clinical pregnancy. Prenatal diagnosis in the second trimester confirmed that the fetus did not carry heterozygous PKD1 gene c.2098-2A>G mutation. Conclusions Our study is the first PGT report targeting the PKD1 gene c.2098-2A>G mutation,which extends the mutation spectrum of PKD1 gene and provides a new line of thinking about the molecular diagnosis and genetic counseling for ADPKD. Combining NGS-based SNP haplotyping for PGT-M with invasive prenatal diagnosis is an effective approach to block the vertical transmission of ADPKD and can be applied to prevent other monogenic genetic diseases.