IVF Center, Department of Obstetrics and Gynecology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, Guangdong, China.
MGI, BGI-Shenzhen, Shenzhen, 518083, Guangdong, China.
J Assist Reprod Genet. 2020 Oct;37(10):2525-2533. doi: 10.1007/s10815-020-01921-4. Epub 2020 Aug 11.
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.
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.
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.
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.
探索一种新的基于全基因组测序（WGS）的植入前遗传学检测（PGT）方法，用于检测新发突变（DNM）与染色体平衡易位的结合。
两个家庭，一个是由 TRPV3 中的 DNM（c.1246C>T）引起的母体 Olmsted 综合征和父亲的罗伯逊易位，另一个是由 FBN1 中的 DNM（c.4952_4955delAATG）引起的父亲 Marfan 综合征和母亲的相互易位，接受了单基因疾病（PGT-M）、染色体非整倍体和结构重排的 PGT。对胚胎和家庭成员进行 WGS 分析。基于梯度测序深度进行生物信息学分析，并进行 DNM 诊断的父母-胚胎单倍型分析。使用羊水样本进行 Sanger 测序、核型分析和染色体微阵列分析，以确认 PGT 结果。
Olmsted 综合征家族的 2 个胚胎在 1 个 PGT 周期后，WGS 显示无 DNMs 的整倍体胚胎；在 2 个周期后，Marfan 综合征家族的 11 个胚胎仅显示 1 个无 DNM、拷贝数变异（CNVs）或非整倍体的正常胚胎。此外，Marfan 综合征家族的 1 个囊胚被移植回子宫；PGT 证实了羊膜穿刺术的检测结果，一个健康的婴儿出生。
基于父母-胚胎单倍型的 WGS 是一种有效的 PGT 策略，用于检测与染色体平衡易位结合的 DNM。我们的结果表明，这是一种可靠有效的诊断方法，对 PGT 患者的 DNM 具有临床应用价值。
