scholarly journals Identification of Novel Biallelic TLE6 Variants in Female Infertility With Preimplantation Embryonic Lethality

2021 ◽  
Vol 12 ◽  
Author(s):  
Manyu Zhang ◽  
Chunyu Liu ◽  
Beili Chen ◽  
Mingrong Lv ◽  
Huijuan Zou ◽  
...  
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Essential Role ◽  
Bioinformatics Analysis ◽  
Oocyte Donation ◽  
Embryonic Lethality ◽  
Female Infertility ◽  
Whole Exome ◽  
Enhancer Of Split ◽  
Lower Expression

Preimplantation embryonic lethality is a rare cause of primary female infertility. It has been reported that variants in the transducin-like enhancer of split 6 (TLE6) gene can lead to preimplantation embryonic lethality. However, the incidence of TLE6 variants in patients with preimplantation embryonic lethality is not fully understood. In this study, we identified four patients carrying novel biallelic TLE6 variants in a cohort of 28 patients with preimplantation embryonic lethality by whole-exome sequencing and bioinformatics analysis, accounting for 14.29% (4/28) of the cohort. Immunofluorescence showed that the TLE6 levels in oocytes from patients were much lower than in normal control oocytes, suggesting that the variants result in the lower expression of the TLE6 protein in oocytes. In addition, a retrospective analysis showed that the four patients underwent a total of nine failures of in vitro fertilization and intracytoplasmic sperm injection attempts, and one of them became pregnant on the first attempt using donated oocytes. Our study extends the genetic spectrum of female infertility caused by variants in TLE6 and further confirms previously reported findings that TLE6 plays an essential role in early embryonic development. In such case, oocyte donation may be the preferred treatment.

scholarly journals Novel Biallelic TLE6 Variants Induce Preimplantation Embryonic Lethality That Cannot be Rescued by IVF or ICSI

2021 ◽  
Author(s):  
Manyu Zhang ◽  
Chunyu Liu ◽  
Beili Chen ◽  
Mingrong Lv ◽  
Huijuan Zou ◽  
...  
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Oocyte Donation ◽  
Embryonic Lethality ◽  
Compound Heterozygous ◽  
Bioinformatics Analyses ◽  
Whole Exome ◽  
Maternal Effect Gene ◽  
Early Embryos ◽  
Oocyte Cytoplasm

Abstract Background: Preimplantation embryonic lethality is a rare cause of primary female infertility. Transducin-like enhancer of split 6 (TLE6) is a maternal effect gene which encodes a component of the subcortical maternal complex located in the oocytes and early embryos. It has been reported that biallelic variants in TLE6 can lead to preimplantation embryonic lethality. However, the incidence of TLE6 variants in patients with preimplantation embryonic lethality is not fully understood.Methods: Whole-exome sequencing and bioinformatics analyses were used to analyze a cohort of 28 patients with preimplantation embryonic lethality. We retrospectively analyzed the process and outcome of their attempts at in vitro fertilization and intracytoplasmic sperm injection. Results: In this study, four patients (14.29 %, 4/28) from three unrelated families in a cohort of 28 individuals with preimplantation embryonic lethality were identified as carrying biallelic TLE6 variants, including two homozygous variants and one compound heterozygous variant. These novel frameshift variants in TLE6 caused truncation of the TLE6 protein likely impairing its function. Immunofluorescence staining of oocytes for TLE6 indicated that it is localized in the oocyte cytoplasm, and that TLE6 was almost absent in the oocytes of the patients carrying biallelic TLE6 variants compared with normal control oocytes. A retrospective analysis showed that the four affected individuals underwent a total of nine in vitro fertilization and intracytoplasmic sperm injection attempts, in which a total of 119 metaphase II oocytes were retrieved, but none of them obtained high-quality blastocysts or obtained a successful pregnancy. However, one of these patients became pregnant on the first attempt using donated oocytes.Conclusions: Biallelic TLE6 variants cause preimplantation embryonic lethality that cannot be rescued by in vitro fertilization or intracytoplasmic sperm injection. Thus, oocyte donation may be the preferred treatment for patients harboring biallelic TLE6 variants.

Mutations in ZP4 are associated with abnormal zona pellucida and female infertility

2021 ◽  
pp. jclinpath-2020-207170
Author(s):  
Xiaoli Wei ◽  
Youzhu Li ◽  
Qicai Liu ◽  
Wensheng Liu ◽  
Xiaohong Yan ◽  
...  
Keyword(s):  
Embryo Development ◽  
Zona Pellucida ◽  
Intracytoplasmic Sperm Injection ◽  
Treatment Strategies ◽  
Suction Pressure ◽  
Human Female ◽  
Female Reproduction ◽  
Efficient Treatment ◽  
Whole Exome

BackgroundThe zona pellucida (ZP) of human oocytes plays essential protective roles in sperm–egg interactions during fertilisation and embryo development. ZP4-null female rabbits exhibit a thin and irregular ZP, which severely impairs embryo development and fertility. However, the effects of ZP4 defect on human female reproduction remain unknown.Methods and resultsWe performed whole-exome sequencing in 26 female patients with abnormal (thin and irregular) ZP and identified heterozygous variants in ZP4 (OMIM: 613514) from 3 patients (approximately 11%). No ZP4 variant was found in the 30 control women with proven fertility. We constructed ZP4-mutated plasmids and found that the variants reduced the secretion of ZP4 in vitro. Lower suction pressure facilitated egg retrieval, and intracytoplasmic sperm injection (ICSI) was a desirable treatment for ZP4-mutated patients with abnormal ZP.ConclusionsWe identified ZP4 as a novel gene for human abnormal ZP and found that lower suction pressure and ICSI are efficient treatment strategies.

scholarly journals Homozygous mutations in REC114 cause female infertility characterised by multiple pronuclei formation and early embryonic arrest

2019 ◽  
Vol 57 (3) ◽  
pp. 187-194 ◽  
Author(s):  
Wenjing Wang ◽  
Jie Dong ◽  
Biaobang Chen ◽  
Jing Du ◽  
Yanping Kuang ◽  
...  
Keyword(s):  
Missense Mutation ◽  
Splice Site ◽  
Genetic Factors ◽  
Splice Site Mutation ◽  
Female Infertility ◽  
Site Mutation ◽  
Whole Exome ◽  
Pronuclear Formation ◽  
Embryonic Arrest

BackgroundAbnormal pronuclear formation during fertilisation and subsequent early embryonic arrest results in female infertility. In recent years, with the prevalence of assisted reproductive technology, a few genes have been identified that are involved in female infertility caused by abnormalities in oocyte development, fertilisation and embryonic development. However, the genetic factors responsible for multiple pronuclei formation during fertilisation and early embryonic arrest remain largely unknown.ObjectiveWe aim to identify genetic factors responsible for multiple pronuclei formation during fertilisation or early embryonic arrest.MethodsWhole-exome sequencing was performed in a cohort of 580 patients with abnormal fertilisation and early embryonic arrest. Effects of mutations were investigated in HEK293T cells by western blotting and immunoprecipitation, as well as minigene assay.ResultsWe identified a novel homozygous missense mutation (c.397T>G, p.C133G) and a novel homozygous donor splice-site mutation (c.546+5G>A) in the meiotic gene REC114. REC114 is involved in the formation of double strand breaks (DSBs), which initiate homologous chromosome recombination. We demonstrated that the splice-site mutation affected the normal alternative splicing of REC114, while the missense mutation reduced the protein level of REC114 in vitro and resulted in the loss of its function to protect its partner protein MEI4 from degradation.ConclusionsOur study has identified mutations in REC114 responsible for human multiple pronuclei formation and early embryonic arrest, and these findings expand our knowledge of genetic factors that are responsible for normal human female meiosis and fertility.

scholarly journals Homozygous variants in SYCP2L cause premature ovarian insufficiency

2020 ◽  
pp. jmedgenet-2019-106789 ◽  
Author(s):  
Wen-Bin He ◽  
Chen Tan ◽  
Ya-Xin Zhang ◽  
Lan-Lan Meng ◽  
Fei Gong ◽  
...  
Keyword(s):  
Bioinformatic Analysis ◽  
Female Infertility ◽  
Premature Ovarian Insufficiency ◽  
Loss Of Function ◽  
Ovarian Insufficiency ◽  
Genetic Causes ◽  
Functional Characterisation ◽  
Whole Exome ◽  
Functional Analyses

BackgroundThe genetic causes of the majority of cases of female infertility caused by premature ovarian insufficiency (POI) are unknown.ObjectiveTo identify the genetic causes of POI in 110 patients.MethodsWhole-exome sequencing was performed on 110 patients with POI, and putative disease-causative variants were validated by Sanger sequencing. Bioinformatic and in vitro functional analyses were performed for functional characterisation of the identified candidate disease-causative variants.ResultsWe identified two homozygous variants (NM_001040274: c.150_151del (p.Ser52Profs*7), c.999A>G (p.Ile333Met)) in SYCP2L in two patients, which had co-segregated with POI in these families. Bioinformatic analysis predicted that the two variants are deleterious, and in vitro functional analysis showed that mutant SYCP2L proteins exhibited mislocalisation and loss of function.ConclusionsSYCP2L is a novel gene found to be responsible for human POI. Our findings provide a potential molecular marker for POI and improve the understanding of the genetic basis of female infertility.

scholarly journals OR24-06 USP8 Genetic Variants May Contribute to the Development of Bilateral Adrenal Hyperplasia and ACTH-Independent Cushing Syndrome

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Nikolaos Settas ◽  
Eva Kassi ◽  
Anna Angelousi ◽  
Fabio Rueda Faucz ◽  
Constantine A Stratakis
Keyword(s):  
Cushing Syndrome ◽  
Cushing Disease ◽  
Control Tissue ◽  
Whole Exome ◽  
Bilateral Adrenal Hyperplasia ◽  
Tumor Level ◽  
The Right ◽  
Canonical Wnt ◽  
Lower Expression

Abstract Background: Bilateral adrenocortical hyperplasias (BAHs), including primary pigmented nodular adrenocortical disease (PPNAD), isolated micronodular adrenocortical disease (iMAD) and primary macronodular adrenocortical hyperplasia (PMAH), are rare causes of ACTH-independent Cushing syndrome (CS). PPNAD and iMAD usually present in children or adolescents as multiple small (<1cm), cortisol-producing adrenocortical nodules. On the other hand, PMAH is most frequently identified in older patients with multiple large adrenal nodules. Most patients with PPNAD have PRKAR1A mutations whereas patients with PMAH may harbor variants in other genes (ARMC5, MC2R, GNAS, APC, MEN1). Even though several genes have been associated with ACTH-independent CS, there are still cases that the genetic cause has not been elucidated. Clinical cases: Herein, we present two unrelated patients with ACTH-independent CS that harbor USP8 gene variants. USP8 is mainly known for being mutated in Cushing disease but as a deubiquitinase it may be involved into the Wnt/β-Catenin signaling pathway. The first patient was diagnosed with BAH on prenatal ultrasound (26 gestational week) and subsequently required bilateral adrenalectomy for CS as she had virilization, hirsutism, hypertension and cardiac hypertrophy 9 weeks old. Adrenalectomy revealed that she had iMAD. She also presented with hemihypertrophy of the right leg, labia and mild newborn hypoglycemia, however she was negative for Beckwith-Wiedemann mutation. Gene analysis of PRKAR1A did not reveal any mutations. After whole exome sequencing (WES), we found a novel heterozygous USP8 variant (c.1387_1393delinsT, p.Ala463_Ile465delinsPhe) at germline level and loss of heterozygosity (LOH) at tumor level. Immunohistochemistry showed significantly lower expression of USP8 protein in both of her adrenals compared to a control tissue. The second case is a 59-year old female with osteoporosis who failed to suppress cortisol levels after low dose dexamethasone administration. MRI revealed an adenoma on the right adrenal (2.6cm). She underwent right adrenalectomy and was found to have PMAH. We performed WES in germline level and we detected a novel heterozygous missense USP8 variant (c.287A>G, p.Lys96Arg) that is present also at tumor level. Immunohistochemistry showed significantly lower expression of USP8 protein in her adrenal tumor compared to the control tissue. No LOH was identified. Conclusion: This is the first report of the association of USP8 in ACTH-independent CS and the preliminary findings support UPS8 involvement in the development of adrenocortical disease. We are currently performing further in vitro studies to evaluate the effect of these two USP8 variants into the canonical Wnt pathway which is commonly involved in adrenocortical disorders.

scholarly journals Compound heterozygous ZP1 mutations cause empty follicle syndrome in infertile sisters

2018 ◽  
Author(s):  
Ling Sun ◽  
Xiang Fang ◽  
Zhi-Heng Chen ◽  
Han-Wang Zhang ◽  
Xiao-Fang Peng ◽  
...  
Keyword(s):  
Medical Treatment ◽  
Zona Pellucida ◽  
Ovarian Stimulation ◽  
Bioinformatics Analysis ◽  
Compound Heterozygous ◽  
The Core ◽  
Empty Follicle Syndrome ◽  
Whole Exome ◽  
Compound Heterozygous Variants

AbstractPurposeEmpty follicle syndrome (EFS) is a condition in which no oocyte is retrieved from mature follicles after proper ovarian stimulation in an in vitro fertilization (IVF) procedure. Genetic evidence accumulates for the etiology of recurrent EFS even with improved medical treatment which had avoided the pharmacological or iatrogenic problems. Here, this study investigated the genetic cause of recurrent EFS in a family with two infertile sisters.MethodsIn this work, we present two infertile sisters in a family with recurrent EFS after three cycles of standard ovarian stimulation with hCG and/or GnRHa therapy. We performed whole-exome sequencing and targeted sequencing in the core members of this family, and further bioinformatics analysis to identify pathogenesis of gene.ResultsWe identified compound heterozygous variants, c.161_165del (p.54fs) and c.1166_1173del (p.389fs), on zona pellucida glycoprotein 1 (ZP1) gene, which were shared with two infertile sisters. Cosegregation tests on the affected and unaffected members of this family confirmed that the allelic mutants were transmitted from either parent.ConclusionsThis EFS phenotype was distinct from the previously reported disruption of zona pellucida due to homozygous ZP1 defects. We thus propose that the specific mutations in ZP1 gene may render a causality for the recurrent EFS.

Rare Germline GLMN Variants Identified from Blue Rubber Bleb Nevus Syndrome Might Impact mTOR Signaling

2020 ◽  
Vol 22 (10) ◽  
pp. 675-682 ◽  
Author(s):  
Jie Yin ◽  
Zhongping Qin ◽  
Kai Wu ◽  
Yufei Zhu ◽  
Landian Hu ◽  
...  
Keyword(s):  
Gastrointestinal Tract ◽  
Sanger Sequencing ◽  
Somatic Mutations ◽  
Venous Malformation ◽  
Underlying Disease ◽  
Mtor Signaling ◽  
Functional Effect ◽  
Germline Variants ◽  
Whole Exome

Backgrounds and Objective: Blue rubber bleb nevus syndrome (BRBN) or Bean syndrome is a rare Venous Malformation (VM)-associated disorder, which mostly affects the skin and gastrointestinal tract in early childhood. Somatic mutations in TEK have been identified from BRBN patients; however, the etiology of TEK mutation-negative patients of BRBN need further investigation. Method: Two unrelated sporadic BRBNs and one sporadic VM were firstly screened for any rare nonsilent mutation in TEK by Sanger sequencing and subsequently applied to whole-exome sequencing to identify underlying disease causative variants. Overexpression assay and immunoblotting were used to evaluate the functional effect of the candidate disease causative variants. Results: In the VM case, we identified the known causative somatic mutation in the TEK gene c.2740C>T (p.Leu914Phe). In the BRBN patients, we identified two rare germline variants in GLMN gene c.761C>G (p.Pro254Arg) and c.1630G>T(p.Glu544*). The GLMN-P254R-expressing and GLMN-E544X-expressing HUVECs exhibited increased phosphorylation of mTOR-Ser-2448 in comparison with GLMN-WTexpressing HUVECs in vitro. Conclusion: Our results demonstrated that rare germline variants in GLMN might contribute to the pathogenesis of BRBN. Moreover, abnormal mTOR signaling might be the pathogenesis mechanism underlying the dysfunction of GLMN protein.

Sign in / Sign up

Export Citation Format

Share Document