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Novel application of luciferase assay for the in vitro functional assessment of KAL1 variants in three females with septo-optic dysplasia (SOD)

Abstract

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.

Type Journal
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.;
Responsible Garvan Author Dr Mark McCabe
Publisher Name MOLECULAR AND CELLULAR ENDOCRINOLOGY
Published Date 2015-01-01
Published Volume 417
Published Pages 63-72
Status Published in-print
URL link to publisher's version http://www.ncbi.nlm.nih.gov/pubmed/26375424
OpenAccess link to author's accepted manuscript version https://publications.gimr.garvan.org.au/open-access/13224