Clinical Characterization and Molecular Profiling by Targeted Next-Generation Sequencing in a Large Indian Cohort...

46,XY differences in sex development (DSD) comprise a heterogeneous group of conditions. Molecular diagnosis guides management by giving insights into the pathophysiology of sex development, reproduction, tumour formation and extragenital issues.

Children with 46,XY DSD from a tertiary hospital in India underwent comprehensive assessment and stepwise genetic testing. Targeted testing for SRD5A2 and AR was performed in cases with suspected 5α-reductase type 2 (5αR2) deficiency/androgen insensitivity syndrome (AIS). For all other patients, and those without variants in SRD5A2 or AR on Sanger sequencing, NGS using a targeted 155-gene panel was performed. Longitudinal clinical data was also collated.

One hundred and forty-seven children with 46,XY DSD with a median (interquartile range) age of 3.8 (1.4, 10.6) years were enrolled. Provisional clinical diagnoses were 5αR2 deficiency/AIS (n = 83, 56.5%), gonadal dysgenesis (n = 31, 21%), testosterone biosynthetic defect (n = 11, 7.5%) and others (n = 22, 15%) based on clinical, biochemical and radiological assessment. Sequential single gene testing for SRD5A2 and AR performed in 75 patients with a clinical diagnosis of 5αR2 deficiency/AIS identified pathogenic/likely pathogenic variants in 44 subjects. NGS in the remaining 103 children revealed pathogenic/likely pathogenic variants in 20 subjects across 12 genes, with NR5A1 being the most frequent (7/103). Four subjects had variants of uncertain significance (VUS) deemed possibly pathogenic due to good genotype-phenotype correlation. Overall, 68/147 (46%) achieved a molecular diagnosis in this cohort.

Sanger sequencing followed by NGS could provide molecular diagnosis in 46% of this Indian cohort with 46,XY DSD. SRD5A2, AR and NR5A1 were the most frequently implicated genes.