Defective Function of Inhibitor of κB Kinase Subunit Beta Associated With Multiple Immune-Mediated Disorders.
Kiril Malovitski, Noy Keller Rosenthal, Lubna Khair, David Hagin, Tal Freund, Eylon Sharoni, Alon Peled, Yarden Feller, Rawaa Ishtewy, Janan Mohamad, Ofer Sarig, Liat Samuelov, Eli Sprecher, Mor Pavlovsky
Abstract
Open AccessAbnormal NF-κB activity has been previously implicated in a range of immune-mediated disorders. Here, we aimed to elucidate the genetic basis underlying the co-occurrence of vitiligo, Addison's disease and granuloma annulare in a 43-year-old woman. Whole-exome sequencing identified a heterozygous splice-site variant (c.1364+1G>A, p.Met455fsTer1) in IKBKB, encoding the Inhibitor of κB kinase subunit beta (IKKβ), predicted to result in a premature stop codon. Immunoblotting of keratinocytes transfected with the mutant construct demonstrated the presence of a truncated form of IKKβ. Using a luciferase reporter assay under the control of NF-κB-responsive element, we demonstrated significantly reduced activity of the mutant protein compared to wild-type, supporting a loss-of-function mechanism. In line with this observation, the mutant protein was found to result in decreased expression levels of genes known to be regulated by NF-κB. Furthermore, HeLa cells transfected with the p.Met455fsTer1 variant or IKBKB-targeted siRNA exhibited markedly reduced levels of p105 and its processed form p50, compared with HeLa cells transfected with wild-type IKBKB or control siRNA, respectively. Collectively, these findings indicate that a loss-of-function effect in IKBKB may underlie the co-occurrence of a number of immune-mediated disorders through disruption of NF-κB signalling.