Next‐generation sequence‐based preimplantation genetic testing for monogenic disease resulting from maternal mosaicism

ABSTRACT Background Mosaicism poses challenges for genetic counseling and preimplantation genetic testing for monogenic disorders (PGT‐M). NGS‐based PGT‐M has been extensively used to prevent the transmission of monogenic defects, but it has not been evaluated in the application of PGT‐M resulting from mosaicism. Methods Four women suspected of mosaicism were confirmed by ultra‐deep sequencing. Blastocyst trophectoderm cells and polar bodies were collected for whole genome amplification, followed by pathogenic variants detection and haplotype analysis based on NGS. The embryos free of the monogenic disorders were transplantable. Results Ultra‐deep sequencing confirmed that the four women harbored somatic mosaic variants, with the proportion of variant cells at 1.12%, 9.0%, 27.60%, and 91.03%, respectively. A total of 25 blastocysts were biopsied and detected during four PGT cycles and 5 polar bodies were involved in one cycle additionally. For each couple, a wild‐type embryo was successfully transplanted and confirmed by prenatal diagnosis, resulting in the birth of four healthy infants. Conclusions Mosaic variants could be effectively evaluated via ultra‐deep sequencing, and could be prevented the transmission by PGT. Our work suggested that an NGS‐based PGT approach, involving pathogenic variants detection combined with haplotype analysis, is crucial for accurate PGT‐M with mosaicism. NGS‐based PGT is an effective strategy for the detection of monogenic diseases resulting from mosaicism. Furthermore, NGS‐based PGT combined with PB genetic analysis can improve the accuracy of PGT for monogenic diseases due to maternal mosaicism. This approach will assist clinicians to undertake a more objective assessment of disease recurrence risk for mosaic individuals and obtain an accurate diagnosis of the genotype of embryos for reproductive interventions.