Novel application of luciferase assay for the in vitro functional assessment of KAL1 variants in three females with septo-optic dysplasia (SOD)
KAL1 is implicated in 5% of Kallmann syndrome cases, a disorder which genotypically overlaps with septo-optic dysplasia (SOD). To date, a reporter-based assay to assess the functional consequences of KAL1 mutations is lacking. We aimed to develop a luciferase assay for novel application to functional assessment of rare KAL1 mutations detected in a screen of 422 patients with SOD. Quantitative analysis was performed using L6-myoblasts stably expressing FGFR1, transfected with a luciferase-reporter vector containing elements of the FGF-responsive osteocalcin promoter. The two variants assayed [p.K185N, p.P291T], were detected in three females with SOD (presenting with optic nerve hypoplasia, midline and pituitary defects). Our novel assay revealed significant decreases in transcriptional activity [p.K185N: 21% (p < 0.01); p.P291T: 40% (p < 0.001)]. Our luciferase-reporter assay, developed for assessment of KAL1 mutations, determined that two variants in females with hypopituitarism/SOD are loss-of-function; demonstrating that this assay is suitable for quantitative assessment of mutations in this gene.
|ISBN||1872-8057 (Electronic) 0303-7207 (Linking)|
|Authors||McCabe, M. J.?; Hu, Y.?; Gregory, L. C.?; Gaston-Massuet, C.?; Alatzoglou, K. S.?; Saldanha, J. W.?; Gualtieri, A.?; Thankamony, A.?; Hughes, I.?; Townshend, S.?; Martinez-Barbera, J. P.?; Bouloux, P. M.?; Dattani, M. T.;|
|Publisher Name||MOL CELL ENDOCRINOL|
|Published Date||2015-01-01 00:00:00|