Clinical application of single-sperm-based SNP haplotyping for PGT of monogenic hereditary disease associated with de novo mutations
10.3760/cma.j.cn101441-20220924-00413
- VernacularTitle:单精子测序技术在新发突变单基因遗传病家系胚胎植入前遗传学检测中的应用
- Author:
Xuanyou ZHOU
1
;
Songchang CHEN
;
Shuyuan LI
;
Chenming XU
Author Information
1. 复旦大学附属妇产科医院妇产科遗传中心，上海 200001
- Publication Type:Journal Article
- Keywords:
Preimplantation genetic testing;
Monogenic disease;
Single-sperm-based single-nucleotide polymorphism haplotyping;
Denovo mutation
- From:
Chinese Journal of Reproduction and Contraception
2022;42(11):1127-1135
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the application value of single-sperm-based single-nucleotide polymorphism (SNP) haplotyping in preimplantation genetic testing of monogenic disorders (PGT-M) associated with de novo mutations. Methods:Whole genome amplification (WGA) of the isolated single sperm was performed based on the multiple displacement amplification (MDA). WGA products were tested for the pathogenic mutation site and informative polymorphic SNP loci located within 2M upstream or downstream of the target gene to establish a sperm-based SNP haplotype. Biopsy samples obtained from embryos were subjected to WGA and next-generation sequencing (NGS). All embryos were verified via haplotype analysis and normal embryos were selected for transfer.Results:Totally 16 sperm samples were selected. Haplotypes of the affected male in 3 families with monogenic hereditary diseases including primary hyperoxaluria type 1 (PH1), Kabuki syndrome and Epidermolysis bullosa (EB) were successfully constructed using single sperm cell sequencing. PGT-M results showed that there were 10 embryos carrying paternal pathogenic variations, while the other 6 embryos did not carry paternal pathogenic variations, 2 of them had chromosomal copy number variations (CNVs). Four embryos obtained from 2 families were found to be normal after NGS according to single-sperm-based SNP haplotype analysis. However, no successful pregnancy was obtained.Conclusion:For males carrying de novo mutations, single-sperm-based SNP haplotyping can be applied for PGT to construct paternal haplotype.
