De Novo Paternal FBN1 Mutation Detected in Embryos Before Implantation

Wang, S; Niu, Zhendong; Wang, H; Ma, Ming; Zhang, W; S, Fang Wang; Wang, J; H, Yan; Y, Liu; Duan, Naibin; Zhang, X; Yao, Yuanqing

doi:10.12659/msm.904546

Cited by 4 publications

(6 citation statements)

References 38 publications

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“…The diagnostic accuracy may be affected when using STR-based haplotype linkage analysis if the recombination occurs between STR loci and target genes. Currently, SNP-based linkage analysis is increasingly applied to PGT-M because SNPs are more widely distributed and exist at higher densities in the genome than STRs ( Wang et al, 2017 ).…”

Section: Discussionmentioning

confidence: 99%

“…However, this approach also has obvious drawbacks. For example, it is difficult to successfully construct a haplotype if the patient has a poor ovarian reserve, with insufficiently mature oocytes ( Wang et al, 2017 ; Chen et al, 2019 ; Yuan et al, 2020 ; Huang et al, 2022 ). The second category uses affected embryos as probands in the linkage phase and analyzes them via NGS.…”

Section: Discussionmentioning

confidence: 99%

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Establishment of linkage phase, using Oxford Nanopore Technologies, for preimplantation genetic testing of Coffin-Lowry syndrome with a de novo RPS6KA3 mutation

Wen,

Du,

et al. 2023

Front. Genet.

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Background: This study aimed to perform preimplantation genetic testing (PGT) for a female Coffin-Lowry Syndrome (CLS) patient with a de novo mutation (DNM) in RPS6KA3. It was challenging to establish the haplotype in this family because of the lack of information from affected family members. Hence, we explored a new and reliable strategy for the detection of the DNM in PGT, using Oxford Nanopore Technologies (ONT) and the MARSALA platform.Methods: We performed whole-exome sequencing (WES) on the proband and confirmed the pathogenic mutation by Sanger sequencing. The proband then underwent PGT to prevent the transmission of the pathogenic mutation to her offspring. We diverged from the conventional methods and used long-read sequencing (LRS) on the ONT platform to directly detect the mutation and nearby SNPs, for construction of the haplotype in the preclinical phase of PGT. In the clinical phase of embryo diagnosis, the MARSALA method was used to detect both the SNP-based haplotype and chromosome copy number variations (CNVs), in each blastocyst. Finally, a normal embryo was selected by comparison to the haplotype of the proband and transferred into the uterus. Sanger sequencing and karyotyping were performed by amniocentesis, at 17 weeks of gestation, to confirm the accuracy of PGT.Results: Using WES, we found the novel, heterozygous, pathogenic c.1496delG (p.Gly499Valfs*25) mutation of RPS6KA3 in the proband. The SNP-based haplotype that was linked to the pathogenic mutation site was successfully established in the proband, without the need for other family members to be tested with ONT. Eight blastocysts were biopsied to perform PGT and were assessed with a haplotype linkage analysis (30 SNP sites selected), to give results that were consistent with direct mutation detection using Sanger sequencing. The results of PGT showed that three of the eight blastocysts were normal, without the DNM. Moreover, the patient had a successful pregnancy, after transfer of a normal blastocyst into the uterus, and delivered a healthy baby.Conclusion: The ONT platform, combined with the MARSALA method, can be used to perform PGT for DNM patients without the need for other samples as a reference.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Establishment of linkage phase, using Oxford Nanopore Technologies, for preimplantation genetic testing of Coffin-Lowry syndrome with a de novo RPS6KA3 mutation

Wen,

Du,

et al. 2023

Front. Genet.

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“…Diagnosis of MFS relies on family history and multi-system scores. Due to the clinical heterogeneity and diversification of mutations, it is difficult for doctors to diagnose mild cases or prenatal patients 171 . However, with the advancements in genetic testing, this problem is expected to be solved.…”

Section: Discussionmentioning

confidence: 99%

The Molecular Genetics of Marfan Syndrome

Du¹,

Zhang²,

Zhuang³

et al. 2021

Int. J. Med. Sci.

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Marfan syndrome (MFS) is a complex connective tissue disease that is primarily characterized by cardiovascular, ocular and skeletal systems disorders. Despite its rarity, MFS severely impacts the quality of life of the patients. It has been shown that molecular genetic factors serve critical roles in the pathogenesis of MFS. FBN1 is associated with MFS and the other genes such as FBN2 , transforming growth factor beta (TGF-β) receptors ( TGFBR1 and TGFBR2 ), latent TGF-β-binding protein 2 ( LTBP2 ) and SKI , amongst others also have their associated syndromes, however high overlap may exist between these syndromes and MFS. Abnormalities in the TGF-β signaling pathway also contribute to the development of aneurysms in patients with MFS, although the detailed molecular mechanism remains unclear. Mutant FBN1 protein may cause unstableness in elastic structures, thereby perturbing the TGF-β signaling pathway, which regulates several processes in cells. Additionally, DNA methylation of FBN1 and histone acetylation in an MFS mouse model demonstrated that epigenetic factors play a regulatory role in MFS. The purpose of the present review is to provide an up-to-date understanding of MFS-related genes and relevant assessment technologies, with the aim of laying a foundation for the early diagnosis, consultation and treatment of MFS.

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“…Subsequently, targeted sequencing by NGS was developed, which detected the DNM in FBN1 for PGT-M. However, this required the assistance of a customized probe in the target region and did not detect CNVs and aneuploidy [9]. Moreover, three biopsied blastocysts in this study were considered wild-type homozygotes in the DNM site of FBN1, while the haplotypes were not verified by single-sperm detection and could not exclude ADO in this site which may result in misdiagnoses in the PGT.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, single-nucleotide polymorphism (SNP) array and nextgeneration sequencing (NGS) techniques applied in PGT have enabled more precise diagnosis not only because more genetic markers (such as SNPs) were used instead of the previously used short tandem repeats (STRs), to construct haplotypes, but also because genetic diseases and aneuploidy could be identified concurrently [7,8]. In addition, other studies reported the direct use of genetic information from parent-linked-embryos to phase haplotypes with preliminary target sequencing, but the diagnosis was customized and unavailable for chromosomes [9].…”

Section: Introductionmentioning

confidence: 99%

A whole-genome sequencing–based novel preimplantation genetic testing method for de novo mutations combined with chromosomal balanced translocations

Yuan

Xia

et al. 2020

J Assist Reprod Genet

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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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De Novo Paternal FBN1 Mutation Detected in Embryos Before Implantation

Cited by 4 publications

References 38 publications

Establishment of linkage phase, using Oxford Nanopore Technologies, for preimplantation genetic testing of Coffin-Lowry syndrome with a de novo RPS6KA3 mutation

Establishment of linkage phase, using Oxford Nanopore Technologies, for preimplantation genetic testing of Coffin-Lowry syndrome with a de novo RPS6KA3 mutation

The Molecular Genetics of Marfan Syndrome

A whole-genome sequencing–based novel preimplantation genetic testing method for de novo mutations combined with chromosomal balanced translocations

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