Abstract
Purpose
To explore a new preimplantation genetic testing (PGT) method for de novo mutations (DNMs) combined with chromosomal balanced translocations by whole-genome sequencing (WGS) using the MGISEQ-2000 sequencer.
Methods
Two families, one with maternal Olmsted syndrome caused by DNM (c.1246C>T) in TRPV3 and a paternal Robertsonian translocation and one with paternal Marfan syndrome caused by DNM (c.4952_4955delAATG) in FBN1 and a maternal reciprocal translocation, underwent PGT for monogenetic disease (PGT-M), chromosomal aneuploidy, and structural rearrangement. WGS of embryos and family members were performed. Bioinformatics analysis based on gradient sequencing depth was performed, and parent-embryo haplotyping was conducted for DNM diagnosis. Sanger sequencing, karyotyping, and chromosomal microarray analysis were performed using an amniotic fluid sample to confirm the PGT results.
Results
After 1 PGT cycle, WGS of 2 embryos from the Olmsted syndrome family revealed euploid embryos without DNMs; after 2 cycles, the 11 embryos from the Marfan syndrome family showed only 1 normal embryo without DNM, copy number variations (CNVs), or aneuploidy. Moreover, 1 blastocyst from the Marfan syndrome family was transferred back to the uterus; the amniocentesis test results were confirmed by PGT and a healthy infant was born.
Conclusions
WGS based on parent-embryo haplotypes was an effective strategy for PGT of DNMs combined with a chromosomal balanced translocation. Our results indicate this is a reliable and effective diagnostic method that is useful for clinical application in PGT of patients with DNMs.
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Acknowledgments
The authors thank all patients and their family members for their participation in this study.
Funding
This work was partially supported by the National Natural Science Foundation of China (No. 81801431), the Chinese Medical Association clinical medical research special fund, Research and development of young physicians in reproductive medicine (No. 18010060735), the Shenzhen Birth Defect Screening Project Lab (JZF No. [2016] 750), and the Shenzhen Municipal Government of China (No. JCYJ20170412152854656).
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Fig. S1
Sanger sequencing results of the Olmsted syndrome family, their 2 embryos, and related the first polar body (PB1) and the second polar body (PB2) for detecting maternal missense mutation (c.1246C > T) in the TRPV3 gene. (PNG 134 kb)
Figure S2
Sanger sequencing results of the Marfan syndrome family and their 11 embryos for detecting paternal frameshift mutation (c.4952_4955delAATG) in the FBN1 gene. (PNG 221 kb)
Fig. S3
Validations of the fetus in the Marfan syndrome family. (a) Sanger sequencing of the amniotic fluid cells confirms that the baby does not have the mutant allele in TRPV3. (b) Chromosomal karyotype analysis reveals that the fetus has the same karyotype as the mother who carries the chromosomal reciprocal translocation (46, XN, t (10; 20) (q26.1; q13.1)). (c) Single nucleotide polymorphism (SNP) array for the detection of copy number variations (CNVs) and aneuploidy shows that the fetus has no chromosomal abnormalities. (PNG 3208 kb)
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Yuan, P., Xia, J., Ou, S. et al. A whole-genome sequencing–based novel preimplantation genetic testing method for de novo mutations combined with chromosomal balanced translocations. J Assist Reprod Genet 37, 2525–2533 (2020). https://doi.org/10.1007/s10815-020-01921-4
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DOI: https://doi.org/10.1007/s10815-020-01921-4