The importance of the multiplex ligation-dependent probe amplification in the identification of a novel two-exon deletion of the NR5A1 gene in a patient with 46,XY differences of sex development

Abstract

Gonadal dysgenesis (GD) is a rare cause of differences of sex development (DSD) with highly variable clinical and genetic conditions. Although identification of the causative genetic alterations can offer a clearer prognosis and personalized management to patients, more than 50% of the DSD cases still do not have an accurate genetic diagnosis. NR5A1 (previously known as SF-1), is a transcriptional regulator of genes required for normal development and functional maintenance of the gonads and the adrenal glands. Nucleotide sequence variants of the NR5A1 gene have been reported in numerous patients with GD with or without adrenal failure, however, microdeletion or partial deletion in the NR5A1 gene have been described only in a few GD cases. In this case study, we present a subject with female phenotype, mild clitoromegaly, partial GD and normal adrenal function. Cytogenetic analysis revealed a 46,XY SRY + karyotype. Microarray analysis did not identify pathogenic copy number variations, nor did panel sequencing of the most common DSD genes. Subsequently, multiplex ligationdependent probe amplification (MLPA) was performed to test for small deletion/duplication of the most frequently affected genes associated with GD. Using this method, we have identified a novel heterozygous deletion involving exons 5 and 6 of the NR5A1 gene as the cause of abnormal sexual development of the patient. This report expands our knowledge about the range and pathogenetic role of NR5A1 mutations associated with partial gonadal dysgenesis in 46,XY DSD. Furthermore, our data emphasises the indispensable role of MLPA in the diagnosis of DSD with unclear etiology.Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Nuclear Receptor Gene Variants Underlying Disorders/Differences of Sex Development through Abnormal Testicular Development

“…According to the Human Gene Mutation Database, more than 200 NR5A1 variants are associated with 46,XY DSD ( Table 1 ). Although most patients have single nucleotide substitutions or small indels, some have structural variants (genetic alterations greater than 50 bp) involving the exons of NR5A1 [ 42 , 43 ]. NR5A1 variants do not affect adrenal functions in most cases; however, several variants have been identified in patients with primary adrenal insufficiency [ 34 , 44 , 45 ].…”

Epididymis cell atlas in a patient with a sex development disorder and a novel NR5A1 gene mutation

“…Heterozygous mutations of the nuclear receptor subfamily 5 group A member 1 ( NR5A1 ) lead to a prevalent 46,XY DSD with a mutation frequency of approximately 15%–20%. 8 …”

Functional study of a novel c.630delG (p.Y211Tfs*85) mutation in NR5A1 gene in a Chinese boy with 46,XY disorders of sex development

“…On the basis of the sex chromosome content, DSD are divided into three categories: sex chromosome DSD; 46,XX DSD; and 46,XY DSD [1]. Heterozygous mutations of NR5A1 are the most prevalent genetic cause in 46,XY DSD individuals, with a frequency of approximately 15-20% [2]. Steroidogenic factor-1 (SF-1), encoded by NR5A1, plays a pivotal role in the adrenal and reproductive development and function as well as in the transcription of genes involved in steroidogenesis [3].…”