Endocrine Abstracts

Premature ovarian insufficiency (POI) is a disorder which affects ~1% of women under 40 years of age. Genetic component has been suggested in the majority of cases of nonsyndromic forms, and recently mutations of NR5A1 have been reported to be associated with POI. In order to evaluate the frequency of NR5A1 mutations in POI together with the functional characterisation of the existing variants, we conducted a genetic study on a large cohort of POI patients.

Patients and method: Hundred and eighty eight patients diagnosed with idiopathic POI and 82 control subjects from the general population were screened for NR5A1 mutations and in vitro functional analysis was performed for the identified variants. HEK293T cells were transiently transfected with either wild type (WT) or mutant NR5A1-containing plasmid. The DNA binding capacity of the variants was evaluated by electrophoretic mobility shift assay (EMSA), while the transcriptional activity was assessed by luciferase assay.

Results: Sequencing NR5A1 gene revealed the presence of four missense variants in three patients (including two missense variants previously described in a patient with POI), nine intronic changes in thirteen patients and six polymorphisms. The three patients with missense variants were 20–33 years old and presented secondary amenorrhea. There was no family history of reproductive disorders, adrenal failure or DSD. Ovarian ultrasonography showed the presence of follicles in two of the three patients. Functional analysis was carried out for three missense variants not present in the control group. EMSA showed no difference in the DNA binding between the studied variants and the WT-NR5A1. Furthermore, no significant differences in the transcriptional activity of the respective variants compared to the WT were found when the luciferase assays was performed.

Conclusions: The results of our study conducted in a large cohort of patients show that NR5A1is not a major cause of idiopathic POI. We suggest that genetic investigation of NR5A1 in POI might be restricted to familial cases.

Declaration of interest: The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project.

Funding: This research did not receive any specific grant from any funding agency in the public, commercial or not-for-profit sector.