New mutations in non-syndromic primary ovarian insufficiency patients identified via whole-exome sequencing

Coloboma and microphthalmia (C/M) are related congenital eye malformations, which can cause significant visual impairment. Molecular diagnosis is challenging as the genes associated to date with C/M account for only a small percentage of cases. Overall, the genetic cause remains unknown in up to 80% of patients. High throughput DNA sequencing technologies, including whole-exome sequencing (WES), are therefore a useful and efficient tool for genetic screening and identification of new mutations and novel genes in C/M. In this study, we analyzed the DNA of 19 patients with C/M from 15 unrelated families using singleton WES and data analysis for 307 genes of interest. We identified seven novel and one recurrent potentially disease-causing variants in CRIM1, CHD7, FAT1, PTCH1, PUF60, BRPF1, and TGFB2 in 47% of our families, three of which occurred de novo. The detection rate in patients with ocular and extraocular manifestations (67%) was higher than in patients with an isolated ocular phenotype (46%). Our study highlights the significant genetic heterogeneity in C/M cohorts and emphasizes the diagnostic power of WES for the screening of patients and families with C/M.

Download Full-text

DMC1 mutation that causes human non-obstructive azoospermia and premature ovarian insufficiency identified by whole-exome sequencing

Journal of Medical Genetics ◽

10.1136/jmedgenet-2017-104992 ◽

2018 ◽

Vol 55 (3) ◽

pp. 198-204 ◽

Cited By ~ 24

Author(s):

Wen-Bin He ◽

Chao-Feng Tu ◽

Qiang Liu ◽

Lan-Lan Meng ◽

Shi-Min Yuan ◽

...

Keyword(s):

Exome Sequencing ◽

Missense Mutation ◽

Whole Exome Sequencing ◽

Male Patient ◽

Histological Analysis ◽

Pedigree Analysis ◽

Obstructive Azoospermia ◽

Ovarian Insufficiency ◽

Causative Gene ◽

Whole Exome

BackgroundThe genetic causes of the majority of male and female infertility caused by human non-obstructive azoospermia (NOA) and premature ovarian insufficiency (POI) with meiotic arrest are unknown.ObjectiveTo identify the genetic cause of NOA and POI in two affected members from a consanguineous Chinese family.MethodsWe performed whole-exome sequencing of DNA from both affected patients. The identified candidate causative gene was further verified by Sanger sequencing for pedigree analysis in this family. In silico analysis was performed to functionally characterise the mutation, and histological analysis was performed using the biopsied testicle sample from the male patient with NOA.ResultsWe identified a novel homozygous missense mutation (NM_007068.3: c.106G>A, p.Asp36Asn) in DMC1, which cosegregated with NOA and POI phenotypes in this family. The identified missense mutation resulted in the substitution of a conserved aspartic residue with asparaginate in the modified H3TH motif of DMC1. This substitution results in protein misfolding. Histological analysis demonstrated a lack of spermatozoa in the male patient’s seminiferous tubules. Immunohistochemistry using a testis biopsy sample from the male patient showed that spermatogenesis was blocked at the zygotene stage during meiotic prophase I.ConclusionsTo the best of our knowledge, this is the first report identifying DMC1 as the causative gene for human NOA and POI. Furthermore, our pedigree analysis shows an autosomal recessive mode of inheritance for NOA and POI caused by DMC1 in this family.

Download Full-text

Whole-exome sequencing identifies GPSM1 as a susceptibility gene for premature ovarian insufficiency

10.21203/rs.3.rs-18442/v1 ◽

2020 ◽

Author(s):

Xuzi Cai ◽

Huijiao Fu ◽

Yan Wang ◽

Qiwen Liu ◽

Xuefeng Wang

Keyword(s):

Exome Sequencing ◽

Whole Exome Sequencing ◽

Caspase 3 ◽

Susceptibility Gene ◽

Primordial Follicle ◽

Antral Follicle ◽

Premature Ovarian Insufficiency ◽

Protein Signaling ◽

Ovarian Insufficiency ◽

Whole Exome

Abstract Background Genetic causes of premature ovarian insufficiency (POI) account for approximately 20~25% of patients. So far, only a few genes have been identified. Results Here, we first identified the c.1840C>A on G-protein signaling modulator 1 (GPSM1) as a susceptibility locus for POI in 10 sporadic POI patients by whole-exome sequencing. The frequency of GPSM1 c.1840C>A was then verified as 3/20 in a POI sample of 20 patients (including the above 10 patients) by Sanger sequencing. RT-PCR and western blot analysis showed the expression of GPSM1 in rat ovaries was increased in the large antral follicle stage compared to the primordial follicle stage (P<0.01). The cell proliferation assay (CCK8) and flow cytometry suggested that the small-interfering RNA-induced silencing of Gpsm1 significantly increased apoptosis and decreased proliferation of rat ovarian granulosa cells (GCs) (P<0.01). Furthermore, suppression of Gpsm1 in GCs reduced levels of cAMP, PKAc, p-CREB as well as the ratio of Bcl-2/Bax, and increased the expression of Caspase-3 and Cleaved Caspase-3 (P<0.01). Conclusions In summary, this study identified a susceptibility variant GPSM1 c.1840C>A of POI for the first time. Gpsm1 was related to rat follicle development, and silencing increased apoptosis and decreased proliferation in rat GCs, possibly through inhibition of the cAMP-PKA-CREB pathway. These findings facilitate the development of the early molecular diagnosis of POI.

Download Full-text

Whole-exome sequencing reveals SALL4 variants in premature ovarian insufficiency: an update on genotype–phenotype correlations

Human Genetics ◽

10.1007/s00439-018-1962-4 ◽

2019 ◽

Vol 138 (1) ◽

pp. 83-92 ◽

Cited By ~ 7

Author(s):

Qiqi Wang ◽

Da Li ◽

Baozhu Cai ◽

Qing Chen ◽

Caihua Li ◽

...

Keyword(s):

Exome Sequencing ◽

Whole Exome Sequencing ◽

Premature Ovarian Insufficiency ◽

Ovarian Insufficiency ◽

Whole Exome

Download Full-text

Whole-exome sequencing in patients with premature ovarian insufficiency: early detection and early intervention

Journal of Ovarian Research ◽

10.1186/s13048-020-00716-6 ◽

2020 ◽

Vol 13 (1) ◽

Author(s):

Hongli Liu ◽

Xiaoli Wei ◽

Yanwei Sha ◽

Wensheng Liu ◽

Haijie Gao ◽

...

Keyword(s):

Exome Sequencing ◽

Whole Exome Sequencing ◽

Ovarian Function ◽

In Silico Analysis ◽

Premature Ovarian Insufficiency ◽

Control Group ◽

Genetic Etiology ◽

Ovarian Insufficiency ◽

Pathogenic Variants ◽

Whole Exome

Abstract Background The loss of ovarian function in women, referred to as premature ovarian insufficiency (POI), is associated with a series of concomitant diseases. POI is genetically heterogeneous, and in most cases, the etiology is unknown. Methods Whole-exome sequencing (WES) was performed on DNA samples obtained from patients with POI, and Sanger sequencing was used to validate the detected potentially pathogenic variants. An in silico analysis was carried out to predict the pathogenicity of the variants. Results We recruited 24 patients with POI and identified variants in POI-related genes in 14 patients, including bi-allelic mutations in DNAH6, HFM1, EIF2B2, BNC, and LRPPRC and heterozygous variants in BNC1, EIF2B4, FOXL2, MCM9, FANCA, ATM, EIF2B3, and GHR. No variants in the above genes were detected in the WES data obtained from 29 women in a control group without POI. Determining a clear genetic etiology could significantly increase patient compliance with appropriate intervention strategies. Conclusions Our study confirmed that POI is a genetically heterogeneous condition and that whole-exome sequencing is a powerful tool for determining its genetic etiology. The results of this study will aid researchers and clinicians in genetic counseling and suggests the potential of WES for the detection of POI and thus early interventions for patients with POI.

Download Full-text