Novel WEE2 compound heterozygous mutations identified in patients with fertilization failure or poor fertilization

Purpose To study associations between novel WEE2 mutations and patients with fertilization failure or poor fertilization. Methods Thirty-one Chinese patients who underwent treatment with assisted reproductive technology and suffered from repeated (at least two times) total fertilization failure (TFF...

Full description

Saved in:

Bibliographic Details
Published in:	Journal of assisted reproduction and genetics 2021-11, Vol.38 (11), p.2861-2869
Main Authors:	Jin, Jiamin, Tong, Xiaomei, Zhang, Yin-Li, Yang, Weijie, Ma, Yerong, Ren, Peipei, Zhou, Feng, Zhang, Songying
Format:	Article
Language:	eng
Subjects:	Adult Cdc2 protein Cell cycle Cell Cycle Proteins - genetics DNA sequencing Female Fertilization Flow cytometry Genetics Gynecology Heterozygote Human Genetics Humans Immunofluorescence Infertility, Female - genetics Infertility, Female - pathology Kinases Localization Male Medicine Medicine & Public Health Mutants Mutation Oocytes Oocytes - metabolism Oocytes - pathology Patients Phosphorylation Pronucleus Protein-Tyrosine Kinases - genetics Proteins Reproductive Medicine Reproductive Techniques, Assisted - statistics & numerical data Reproductive technology Western blotting
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Purpose To study associations between novel WEE2 mutations and patients with fertilization failure or poor fertilization. Methods Thirty-one Chinese patients who underwent treatment with assisted reproductive technology and suffered from repeated (at least two times) total fertilization failure (TFF) or a low fertilization rate were enrolled. Genomic DNA was extracted from patients for whole-exome sequencing. Suspicious mutations were validated by Sanger sequencing. WEE2 protein levels in oocytes from affected patients were examined by immunofluorescence. Disruptive effects of mutations on WEE2 protein stability, subcellular localization, and kinase function were analyzed through western blotting, immunofluorescence, and flow cytometry in HeLa cells. Results Three of thirty-one (9.6%) enrolled patients had six compound heterozygous mutations of the WEE2 gene, and three of them were reported here for the first time (c.115_116insT, c.756_758delTGA, and c.C1459T). Oocytes from affected patients showed decreased WEE2 immunofluorescence signals. In vitro experiments showed that the mutant WEE2 gene caused reduced WEE2 protein levels or cellular compartment translocation in HeLa cells, leading to decreased levels of the phosphorylated Cdc2 protein. Compared with the wild-type WEE2 protein, the mutant WEE2 proteins were also found to have different effects on the cell cycle. Conclusion Three novel compound heterozygous WEE2 variants were found in patients with pronucleus formation failure. This study provides new evidence that WEE2 mutations result in loss of function, which could result in fertilization failure.
ISSN:	1058-0468 1573-7330