AbstractBackgroundRecent genome‐wide association studies of hypospadias have implicated the role of genetic variants in or near the diacylglycerol kinase kappa (DGKK) gene. However, these variants are largely identified among samples of mild and moderate hypospadias cases. Therefore, we evaluated previously identified DGKK variants among second‐ and third‐degree hypospadias cases and controls recruited in Arkansas, a state characterized by a high birth prevalence of hypospadias.MethodsSecond‐ and third‐degree hypospadias non‐Hispanic white cases (n = 36 and n = 9, respectively) and controls (n = 45) were recruited at Arkansas Children's Hospital. Preputial tissue was collected on cases and controls between 2013 and 2017. Cases and controls were genotyped using the Illumina Infinium Global Screening Array. We used logistic regression models to assess the association of genotyped and imputed genetic variants mapped to the DGKK region with second‐ and third‐degree hypospadias.ResultsAll families self‐reported as non‐Hispanic white and genetic principal component analyses did not demonstrate evidence of population stratification. Five DGKK variants previously reported as associated with hypospadias were identified in the genotype data. None of the variants were associated with second‐ or third‐degree hypospadias (range of odds ratios = 0.7–0.9, all p > .05).ConclusionsIn our analyses, genetic variation in DGKK does not play a role in the development of moderate and severe hypospadias. Our findings provide support to the etiologic heterogeneity of hypospadias by all classifications of severity.