Characterization of 35 novel NR5A1/SF-1 variants identified in individuals with atypical sexual development: The SF1next study
Rawda Naamneh Elzenaty, Idoia Martinez de Lapiscina, Chrysanthi Kouri, Kay-Sara Sauter, Grit Sommer, Luis Castaño, Christa E Flück, S Abali, Z Y Abali, F Ahmed, L Akin, M C Almaraz, L Audí, M Aydin, A Balsamo, F Baronio, J Bryce, K Busiah, M Caimari, N Camats-Tarruella, A Campos-Martorell, A Casteràs, S Çetinkaya, Y M Chan, H L Claahsen-van der Grinten, I Costa, M Cools, J H Davies, I Esteva, H Fabbri-Scallet, C A Finlayson, E Garcia, A German, E Globa, G Guerra-Junior, J Guerrero, T Guran, S E Hannema, O Hiort, J Hirsch, I Hughes, M Janner, Z Kolesinska, K Lachlan, D L'Allemand, J K Malikova, M Lang-Muritano, A Lucas-Herald, J Mammadova, K MсElreavey, V Mericq, I Mönig, F Moreno, J Mührer, M Niedziela, A Nordenstrom, B Orman, S Poyrazoglu, J M Rial, M M Rutter, A Rodríguez, T Schafer-Kalkhoff, S Seneviratne, L Tack, R Tadokoro-Cuccaro, A Thankamony, M Tomé, A Vela, M Wasniewska, D Zangen, N Zelinska,- Biochemistry (medical)
- Clinical Biochemistry
- Endocrinology
- Biochemistry
- Endocrinology, Diabetes and Metabolism
Abstract
Context
Steroidogenic factor 1 (NR5A1/SF-1) is a nuclear receptor that regulates sex development, steroidogenesis and reproduction. Genetic variants in NR5A1/SF-1 are common among differences of sex development (DSD) and associate with a wide range of phenotypes, but their pathogenic mechanisms remain unclear.
Objective
Novel, likely disease-causing NR5A1/SF-1 variants from the SF1next cohort of individuals with DSD were characterized to elucidate their pathogenic effect.
Methods
Different in silico tools were used to predict the impact of novel NR5A1/SF-1 variants on protein function. An extensive literature review was conducted to compare and select the best functional studies for testing the pathogenic effect of the variants in a classic cell culture model. The missense NR5A1/SF-1 variants were tested on the promoter luciferase reporter vector -152CYP11A1_pGL3 in HEK293T cells and assessed for their cytoplasmic/nuclear localization by Western blot.
Results
Thirty-five novel NR5A1/SF-1 variants were identified in the SF1next cohort. Seventeen missense NR5A1/SF-1 variants were functionally tested. Transactivation assays showed reduced activity for 40% of the variants located in the DNA binding domain and variable activity for variants located elsewhere. Translocation assessment revealed three variants (3/17) with affected nuclear translocation. No clear genotype-phenotype, structure-function correlation was found.
Conclusions
Genetic analyses and functional assays do not explain the observed wide phenotype of individuals with these novel NR5A1/SF-1 variants. In nine individuals, additional likely disease-causing variants in other genes were found, strengthening the hypothesis that the broad phenotype of DSD associated with NR5A1/SF-1 variants may be caused by an oligogenic mechanism.