PMC10319352_01
PMC10319352_02
Testicular biopsy shows severely impaired spermatogenesis with an aspect of histological mosaicism. The seminiferous tubules were of overall reduced diameter (decreased by approximately 30%-50% compared to physiological adult pubertal seminiferous tubules of 150-250 microm in diameter) and lined by a thickened basal membrane. The seminiferous tubules were either atrophic (*), with Sertoli cells only (**), or presenting spermatogenesis arrest at the spermatocyte stage ( ). The interstitial tissue was fibro-edematous with numerous Leydig cells ( ) (hematoxylin-eosin-saffron). Thick black arrow indicates Sertoli cells, thin black arrow indicates spermatocyte, and solid black triangle indicates spermatogonia. Biopsy fragments were fixed in alcohol, formalin, and acetic acid (AFA) and paraffin-embedded. Sections of 3 mum were stained by hematoxylin-phloxin-saffron. Slide evaluation was performed on a Leica DM2500 microscope. Two different scales: 300 microm (A) and 200 microm (B).
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No signs of malignancy or dysplasia were noticed ( Figure 1 ).
Identification of the NR5A1 variant using Sanger sequencing. Variation was identified using reference NM_004959.5 for NR5A1 transcript on GRCh37/hg19 human genome assembly, NP_004950.2 for SF-1 protein. The screenshot comes from SeqScape 3 software. The nucleotide reference sequence is highlighted in blue. This variant was classified as pathogenic according to the ACMG criteria (PVS1, PM1, and PM2). The polymorphism NM_004959.5: c.437G>C p.(Gly146Ala) was not found. In ACMG criteria: PM, pathogenic moderate; PVS, pathogenic very strong. "PVS1: null variant (non-sense, frameshift, canonical +-1 or 2 splice sites, initiation codon, single or multi-exon deletion) in a gene where loss of function is a known mechanism of disease." "PM1: located in a mutational hot spot and/or critical and well-established functional domain (e.g. active site of an enzyme) without benign variation." "PM2: absent from controls (or at extremely low frequency if recessive) in Exome Sequencing Project, 1000 Genomes or ExAC.".
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At 14 years 10 months of age, after his parents provide signed informed written consent for genetic testing, a molecular analysis of NR5A1 gene (Sanger sequencing on DNA extracted from whole blood) revealed the unreported heterozygous frameshift variant NM_004959.5: c.207del p.(Phe70Serfs*5) (GRCh37/hg19) ( Figure 2 ).