jmedgenet-2017-105191.pdf (1.64 MB)
In vivo bioassay to test the pathogenicity of missense human AIP variants
journal contribution
posted on 2023-06-09, 16:27 authored by Elena Daniela Aflorei, Benjamin Klapholz, Chenghao Chen, Serban Radian, Anca Neluta Dragu, Nina Moderau, Chrisostomos ProdromouChrisostomos Prodromou, Paulo S Ribeiro, Ralf Stanewsky, Marta KorbonitsBackground Heterozygous germline loss-of-function mutations in the aryl hydrocarbon receptor-interacting protein gene (AIP) predispose to childhood-onset pituitary tumours. The pathogenicity of missense variants may pose difficulties for genetic counselling and family follow-up. Objective To develop an in vivo system to test the pathogenicity of human AIP mutations using the fruit fly Drosophila melanogaster. Methods We generated a null mutant of the Drosophila AIP orthologue, CG1847, a gene located on the Xchromosome, which displayed lethality at larval stage in hemizygous knockout male mutants (CG1847exon1_3 ). We tested human missense variants of ‘unknown significance’, with ‘pathogenic’ variants as positive control. Results We found that human AIP can functionally substitute for CG1847, as heterologous overexpression of human AIP rescued male CG1847exon1_3 lethality, while a truncated version of AIP did not restore viability. Flies harbouring patient-specific missense AIP variants (p.C238Y, p.I13N, p.W73R and p.G272D) failed to rescue CG1847exon1_3 mutants, while seven variants (p.R16H, p.Q164R, p.E293V, p.A299V, p.R304Q, p.R314W and p.R325Q) showed rescue, supporting a non-pathogenic role for these latter variants corresponding to prevalence and clinical data. Conclusion Our in vivo model represents a valuable tool to characterise putative disease-causing human AIP variants and assist the genetic counselling and management of families carrying AIP variants.
History
Publication status
- Published
File Version
- Published version
Journal
Journal of Medical GeneticsISSN
0022-2593Publisher
BMJ Publishing GroupExternal DOI
Volume
55Page range
522-529Department affiliated with
- Biochemistry Publications
Full text available
- Yes
Peer reviewed?
- Yes