Abstract
Purpose
To evaluate the unknown genetic causes of teratozoospermia, and determine the pathogenicity of candidate variants.
Methods
A primary infertile patient and his family members were recruited in the West China Second University Hospital of Sichuan University. Whole-exome sequencing was performed to identify causative genes in a man with teratozoospermia. Immunofluorescence staining and western blotting were applied to assess the pathogenicity of the identified variant. Intracytoplasmic sperm injection (ICSI) was used to assist fertilization for the patient with teratozoospermia.
Results
We performed whole-exome sequencing (WES) and detected a novel homozygous frameshift mutation of c.335_336del [p.E112Vfs*3] in DNAJB13 on a primary infertile male patient. Intriguingly, we identified abnormal sperm morphology in this patient, with recurrent respiratory infections and chronic cough. Furthermore, we confirmed that this mutation resulted in negative effects on DNAJB13 expression in the spermatozoa of the affected individual, causing ultrastructural defects in his sperm. Remarkably, our staining revealed that DNAJB13 was expressed in the cytoplasm of primary germ cells and in the flagella of spermatids during spermiogenesis in humans and mice. Finally, we are the first group to report a favorable prognosis using ICSI for a patient carrying this DNAJB13 mutation.
Conclusion
Our study revealed a novel homozygous frameshift mutation of c.335_336del [p.E112Vfs*3] in DNAJB13 involved in teratozoospermia phenotype. Our study greatly expands the spectrum of limited DNAJB13 mutations, and is expected to provide a better understanding of genetic counseling diagnoses and subsequent treatment of male infertility.
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Data availability
Data are available on request from the corresponding author.
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Acknowledgements
We thank the patient and his family for their interest and cooperation. The authors would like to thank Analytical and Testing Center of Sichuan University for the morphology characterization and the authors are grateful to Guiping Yuan for her help of TEM images and Yi He for his help of SEM images.
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Y.S. designed and supervised the study experiments. Y.Y. collected data and conducted the clinical evaluations. J.L. and C.J. performed TEM and SEM. M.L. wrote the first article draft; M.L., Y.Z., and Y.S. performed immunofluorescence staining and Minigene assay. All authors revised and approved the article.
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The study had been approved by the Ethics Committee of West China Second University Hospital, Sichuan University.
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Supplementary Information
Table S1
(XLSX 88 kb)
Fig. S1
Chest X-ray of the normal control and patient. The patient with the DNAJB13 mutant showed no situs -inversus phenotype but increased and disordered lung texture. (JPG 182 kb)
Fig. S2
Results of minigene assay. Pattern diagram showing no harmful splicing effects caused by the DNAJB13 mutation. a Agarose gel electrophoresis of RT-PCR products obtained from wild-type and mutant plasmids. The first channel is a marker, and the second is the empty vector. We observed no change in molecular weight in the mutant plasmid (channel 3) compared with the wildtype plasmid (channel 4). b Proposed model of results by the splice-site mutation. The primers SD6 and SA2 were used to amplify exons 4 and 5 of the DNAHB13 gene. c Sequence analysis of the RT-PCR product obtained from wild-type and mutant plasmid. (PNG 445 kb)
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Liu, M., Li, J., Jiang, C. et al. A novel homozygous mutation in DNAJB13—a gene associated with the sperm axoneme—leads to teratozoospermia. J Assist Reprod Genet 39, 757–764 (2022). https://doi.org/10.1007/s10815-022-02431-1
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DOI: https://doi.org/10.1007/s10815-022-02431-1