研究開発・学会発表

Introduction: Reproductive aging presents a significant obstacle to successful pregnancy, leaving many patients infertile and experiencing recurrent assisted reproductive technology (ART) failures. Approximately 20% of infertility cases remain idiopathic. Recent studies have linked specific gene mutations to fertilization failure and early embryonic arrest. This research focuses on patients with repeated implantation failures and low blastocyst development rates, aiming to identify potential gene mutations that may contribute to compromised embryo development.

Materials & Methods: 25 infertile women who had experienced IVF/ICSI failure more than 5 times, with blastocyst formation rates below 10%, and 10 staff members who had conceived and delivered naturally were enrolled. Informed consent was obtained from all participants. Genomic DNA was extracted from whole blood samples, and whole-exome sequencing was performed. High-impact homozygous variants with allele frequencies ≤0.20 in the 1000 Genome EAS, absent in control subjects with natural pregnancies and deliveries, were identified. The Human Genetic Variation Database (HGVD) was used, and the Hardy-Weinberg equation was applied to evaluate potential candidates for recurrent ART failures caused by genetic mutations.

Results: 57 high-impact homozygous gene mutations were identified, and among them, significant differences in allele frequencies were observed between 8.3KJPN and the patients for ADAM33 (p = 0.009), CEP89 (p = 0.012), CRIPAK (p < 0.001), LGALS9B (p < 0.001), PDZRN3 (p = 0.001), RAET1E (p = 0.007), and SPATA31A3 (p = 0.045). The Hardy-Weinberg equilibrium for each gene based on data from HGVD was calculated. Patients exhibited significantly lower Hardy-Weinberg equilibrium compared to HGVD for ADAM33 (p < 0.001), CEP89 (p = 0.033), OR2T29 (p = 0.003), OR52J3 (p = 0.0497), RABL2A (p < 0.001), RNF17 (p = 0.0499), SPATA31C1 (p = 0.030), and WWTR1 (p < 0.001).Conclusions: In this study, 13 gene mutations associated with repeated ART failures were identified. Among these genes, RNF17, a piRNA pathway gene, and WWTR1, a Hippo signal pathway gene, were the most likely causes of repeated ART failures. These findings may serve as potential diagnostic markers for patients with recurrent ART failures, offering insights into the genetic basis of female infertility.