title: Rare double sex and mab-3-related transcription factor 1 regulatory variants in severe spermatogenic failure.
abstract: The double sex and mab-3-related transcription factor 1 (DMRT1) gene has long been linked to sex-determining pathways across vertebrates and is known to play an essential role in gonadal development and maintenance of spermatogenesis in mice. In humans, the genomic region harboring the DMRT gene cluster has been implicated in disorders of sex development and recently DMRT1 deletions were shown to be associated with non-obstructive azoospermia (NOA). In this work, we have employed different methods to screen a cohort of Portuguese NOA patients for DMRT1 exonic insertions and deletions [by multiplex ligation probe assay (MLPA); n = 68] and point mutations (by Sanger sequencing; n = 155). We have found three novel patient-specific non-coding variants in heterozygosity that were absent from 357 geographically matched controls. One of these is a complex variant with a putative regulatory role (c.-223_-219CGAAA>T), located in the promoter region within a conserved sequence involved in Dmrt1 repression. Moreover, while DMRT1 domains are highly conserved across vertebrates and show reduced levels of diversity in human populations, two rare synonymous substitutions (rs376518776 and rs34946058) and two rare non-coding variants that potentially affect DMRT1 expression and splicing (rs144122237 and rs200423545) were overrepresented in patients when compared with 376 Portuguese controls (301 fertile and 75 normozoospermic). Overall our previous and present results suggest a role of changes in DMRT1 dosage in NOA potentially also through a process of gene misregulation, even though DMRT1 deleterious variants seem to be rare.
authors.affiliation: Graduate Program in Areas of Basic and Applied Biology (GABBA), Abel Salazar Institute of Biomedical Sciences, University of Porto, Porto, Portugal.|Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Porto, Portugal.|Department of Genetics, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: A C
authors.initials: AC
authors.lastname: Lima
authors.affiliation: Department of Genetics, Faculty of Medicine of the University of Porto, Porto, Portugal.
authors.forename: F
authors.initials: F
authors.lastname: Carvalho
authors.affiliation: Department of Human Genetics, National Institute of Health Dr. Ricardo Jorge, Lisboa, Portugal.
authors.forename: J
authors.initials: J
authors.lastname: Gonçalves
authors.affiliation: Department of Genetics, Faculty of Medicine of the University of Porto, Porto, Portugal.
authors.forename: S
authors.identifier: http://orcid.org/0000-0003-1128-833X
authors.initials: S
authors.lastname: Fernandes
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Porto, Portugal.|Abel Salazar Institute of Biomedical Sciences, University of Porto, Porto, Portugal.
authors.forename: P I
authors.initials: PI
authors.lastname: Marques
authors.affiliation: Laboratory of Cell Biology, UMIB, ICBAS, University of Porto, Porto, Portugal.
authors.forename: M
authors.initials: M
authors.lastname: Sousa
authors.affiliation: Department of Genetics, Faculty of Medicine of the University of Porto, Porto, Portugal.
authors.forename: A
authors.initials: A
authors.lastname: Barros
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Porto, Portugal.
authors.forename: S
authors.initials: S
authors.lastname: Seixas
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Porto, Portugal.|Faculty of Sciences of the University of Porto, Porto, Portugal.
authors.forename: A
authors.initials: A
authors.lastname: Amorim
authors.affiliation: Department of Genetics, Washington University School of Medicine, St. Louis, MO, USA.|Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA.
authors.forename: D F
authors.initials: DF
authors.lastname: Conrad
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.|Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Porto, Portugal.
authors.forename: A M
authors.initials: AM
authors.lastname: Lopes
mesh_terms: D053713:Azoospermia / Q000235:genetics*; D001483:Base Sequence; D017353:Gene Deletion; D018628:Gene Dosage / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D011174:Portugal; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016415:Sequence Alignment; D013091:Spermatogenesis / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*
chemical_list: C110547:DMRT1 protein; D014157:Transcription Factors
keywords: alternative splicing; cis-regulatory variants; double sex and mab-3-related transcription factor 1; double sex and mab-3-related transcription factor 1 promoter; non-obstructive azoospermia