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

Abstract

Purpose This study aimed to present the clinical features and gene mutation characteristics of a child with 46,XY disorders of sex development (DSD) caused by a novel heterozygous mutation in the NR5A1 gene to determine the potential association between this heterozygous mutation and the pathogenesis of 46,XY DSD. Methods We present the case of a Chinese child with ambiguous genitalia at birth but a normal adrenal gland. Targeted nextgeneration sequencing, comprising 163 candidate genes involved in sexual differentiation and development, was performed, followed by the functional evaluation of the novel NR5A1 mutation. Result The patient had a novel heterozygous mutation in the NR5A1 gene, c.630delG (p.Y211Tfs*85). Results revealed that overexpression of p.Y211Tfs*85 impaired steroidogenic factor-1 (SF-1) protein synthesis. Immunofluorescence analysis revealed that both SF-1 wild-type and p.Y211Tfs*85 mutation proteins were localized in the cell nucleus. Furthermore, dualluciferase reporter assay results revealed that the p.Y211Tfs*85 mutation could effectively downregulate the transcriptional activation of anti-Müllerian hormone and steroidogenic acute regulatory protein genes (P < 0.01). Additionally, the p.Y211Tfs*85 mutation changed three-dimensional conformation of SF-1, and three conformations could be constructed with the mutated amino acid sequences. Therefore, the novel frameshift mutation could result in decreased protein expression of SF-1. Conclusion We described a novel mutation in NR5A1 and showed that it might affect protein structure, thereby seriously compromising the role of SF-1 in regulating gonadal development. The novel p.Y211Tfs*85 mutation in the NR5A1 gene enriches the boy of information available regarding the mutation spectrum of this gene in the Chinese population. Keywords NR5A1 . SF-1 . AMH . STAR . Disorders of sex development Highlights We identified a patient with a novel heterozygous mutation in the NR5A1 gene, c.630delG (p.Y211Tfs*85). The NR5A1 c.630delG p.Y211Tfs*85 mutation could effectively downregulate the transcriptional activation of the AMH and STAR. This novel frameshift mutation resulted in a decreased protein expression of SF-1. Sinian Pan and Shili Guo contributed equally to this work.

Disorders of Sex Development: Classification, Review, and Impact on Fertility

“…Fabbri et al [82] reported that loss-of-function mutations in the NR5A1 gene can lead to a 46,XY PGD phenotype with mild manifestations, dysgenetic testes, and preserved adrenal function. Similarly, Pan et al [52] reported the clinical characteristics and characteristics of the genetic mutation in an individual with 46,XY DSD caused by a new heterozygous mutation in the NR5A1 gene: c.630delG (p.Y211Tfs*85), presenting the case of a Chinese boy with ambiguous genitalia at birth and normal adrenal glands.…”

“…Similarly, several patients have also been described with a dicentric chromosome for the long arm of the Y chromosome (XYdic) [51], and this is usually associated with mosaicism and with an X0 cell line. Thus, in this group of disorders, various phenotypes can be seen, including normal males, individuals with ambiguous genitalia, male Ps [52], and patients with many of the stigmata of gonadal dysgenesis.…”

Diverse Regulation but Conserved Function: SOX9 in Vertebrate Sex Determination

“…Mutations to SRY account for approximately 15% of 46,XY DSD [ 128 ]; while mutations in other genes such as NR5A1 or SOX9 are known to cause 46,XY DSD as part of a syndrome. To date, a genetic diagnosis is not achieved in around 50% of XY DSD cases [ 128 , 129 ]. Genetic diagnosis of ambiguous genitalia is particularly challenging and a clinical algorithm can facilitate this [ 130 ].…”

A novel c.64G&gt;T (p.G22C) NR5A1 mutation in a Chinese adolescent with 46, XY disorders of sex development: a case report

“…A previous study identi ed a G35E substitution in the DBD region in a patient with 46, XY DSD presenting adrenal insu ciency along with moderately severe gonadal dysplasia, indicating that this variation results in serious adverse effects on NR5A1 function [20]. Additionally, the p.V15M, p.M78I, and p.G91S variations in this region are reportedly responsible for aberrant NR5A1 transcription, with the rst two variations resulting in altered subcellular localization [25]. Moreover, functional studies have shown that the p.S32N, p.N44del, and p.G91D variations in this region reduce the transactivation of cytochrome P450 family 11 subfamily A member 1 [23].…”