Functional assessment of DMRT1 variants and their pathogenicity for isolated male infertility.
DMRT1
gonadal development
luciferase assay
male infertility
nonobstructive azoospermia
Journal
Fertility and sterility
ISSN: 1556-5653
Titre abrégé: Fertil Steril
Pays: United States
ID NLM: 0372772
Informations de publication
Date de publication:
02 2023
02 2023
Historique:
received:
28
06
2022
revised:
18
10
2022
accepted:
19
10
2022
pubmed:
27
12
2022
medline:
8
2
2023
entrez:
26
12
2022
Statut:
ppublish
Résumé
To study the impact of Doublesex and mab-3-related transcription factor 1 (DMRT1) gene variants on the encoded protein's function and the variants' pathogenic relevance for isolated male infertility caused by azoospermia. This study established a novel luciferase assay for DMRT1 missense variants using 2 different target promotors and validated the assay by analyzing previously published variants associated with differences in sex development. University genetics research institute and tertiary referral center for couples' infertility. Eleven infertile men with severely impaired spermatogenesis resulting in crypto- or azoospermia and carrying rare heterozygous missense variants in DMRT1 were identified within the Male Reproductive Genomics study. Luciferase assays with human DMRT1 variants to test functional effects on the CYP19A1 and Stra8 target promoters. We first developed and refined luciferase assays to reliably test the functional impact of DMRT1 missense variants. Next, the assay was validated by analyzing 2 DMRT1 variants associated with differences in sex development, of which c.240G>C p.(Arg80Ser) displayed highly significant effects on both target promoters compared with the wild-type protein (-40% and +100%, respectively) and c.331A>G p.(Arg111Gly) had a significant effect on the Stra8 promoter (-76%). We then systematically characterized 11 DMRT1 variants identified in infertile men. The de novo variant c.344T>A p.(Met115Lys) showed a pronounced loss of function in both DMRT1 target promoters (-100% and -86%, respectively). Variants c.308A>G p.(Lys103Arg) and c.991G>C p.(Asp331His) showed a significant gain of function exclusively for the CYP19A1 promoter (+15% and +19%, respectively). Based on these results, 3 variants were reclassified according to clinical guidelines. The present study highlights the importance of functionally characterizing DMRT1 variants of uncertain clinical significance. Using luciferase assays for diagnostic purposes enables an improved causal diagnosis for isolated male infertility.
Identifiants
pubmed: 36572623
pii: S0015-0282(22)01982-3
doi: 10.1016/j.fertnstert.2022.10.032
pii:
doi:
Substances chimiques
Transcription Factors
0
DMRT1 protein
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
219-228Informations de copyright
Copyright © 2022 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.