Mutational screening of the coding region of growth differentiation factor 9 gene in Indian women with ovarian failure

Objective: To establish the risk associated with mutations in the coding region of GDF9 gene in Indian women with ovarian failure. Design: This case-control study was designed for mutational analysis of the GDF9 coding region in a cohort of women with premature ovarian failure (n = 127), primary amenorrhea (n = 58), and secondary amenorrhea (n = 10) compared with controls (n = 220). Results: This case-control study revealed eight mutations in the GDF9 gene, including five novel mutations: c.1-8C > T, c.199A > C (p.Lys67Glu), c. 205C > T, c.646G > A (p.Val216Mat), and c.1353C > T, and three documented mutations: c.398-39C > G, c.447C > T, and c.546G > A. Missense mutation c. 199A > C was present in 4 of 127 premature ovarian failure (POF) cases and I of 10 secondary amenorrhea cases. The c.646G > A mutation was present in two POF cases. Both missense mutations were absent in controls. Genotype distribution of c.447C > T was significantly different in POF cases than controls (X-2 = 5.93, P = 0.05). We chose two frequent single-nucleotide polymorphisms (c.398-39C > G, c.447C > T) for haplotyping and found that the C-T haplotype was significantly higher in patients (P = 0.03), whereas the C-C haplotype was representative of the control group. Conclusions: We report two rare missense mutations, c.199A > C and c.646G > A, which are associated with ovarian failure. The presence of the c.447 > T mutation might indicate a higher risk for POE Haplotype C-T was significantly associated with ovarian failure, whereas the C-C haplotype was representative of the control group.