Molecular Screening of CYP17A1 and SRD5A2 Gene Alleles Associated with Hypospadias: A Pilot Study from Central India

A BSTRACT Background: Hypospadias is one of the most common congenital anomalies of the male genitalia, with increasing global incidence. The condition is believed to arise from a complex interaction between genetic susceptibility and environmental influences affecting androgen synthesis and signaling during fetal development. Among the genes implicated in this pathway, cytochrome P450 17A1 (CYP17A1) and steroid 5-alpha reductase type 2 (SRD5A2) play critical roles in androgen biosynthesis and conversion, and their polymorphism is associated with hypospadias. However, molecular data from the Indian population remain limited. Objective: The study aimed to detect the associated SRD5A2 and CYP17A1 gene alleles in patients with mid-penile hypospadias and compare them with healthy controls to explore potential genetic markers associated with the condition. Methods: This case–control study was conducted at a tertiary pediatric surgery center in Central India over 20 months. Twenty children with isolated mid-penile hypospadias and 20 age-matched controls were included. Genomic DNA was extracted from peripheral blood samples and subjected to allele-specific polymerase chain reaction (PCR) amplification for targeting CYP17-V89L (rs523349) and SRD5A2-T→C promoter polymorphism (rs743572). Amplified products were analyzed using agarose gel electrophoresis. Results: PCR amplification revealed CYP17-V89L positivity in 20% (4/20) of hypospadias cases and SRD5A2-T→C promoter positivity in 20% (4/20) of cases, while no allele amplification was detected in the control group ( P < 0.05). In addition, 10% (2/20) of patients demonstrated concurrent amplification of both the gene alleles, suggesting possible multigene involvement in androgen pathway disruption. Conclusion: The study demonstrates a potential association between CYP17-V89L and SRD5A2-T→C promoter genetic alterations and mid-penile hypospadias, supporting the role of disrupted androgen biosynthesis in its etiopathogenesis. Larger, detailed genomic studies are needed to further clarify the genetic and environmental factors contributing to hypospadias.