High-Resolution Transcriptional Impact of AIRE: Effects of Pathogenic Variants p.Arg257Ter, p.Cys311Tyr, and Polygenic Risk Variant p.Arg471Cys

Amund Holte Berger^1*Bergithe Eikeland Oftedal¹Anette S. B. Wolff^1,2Eystein Sverre Husebye^1,2Per Morten Knappskog^1,3Eirik Bratland^1,3*Stefan Johansson^1,2,3,4*

• ¹Department of Clinical Science, University of Bergen, Bergen, Norway
• ²Department of Medicine, Haukeland University Hospital, Bergen, Hordaland, Norway
• ³Center for Medical Genetics and Molecular Medicine, Haukeland University Hospital, Bergen, Hordaland, Norway
• ⁴Children and Youth Clinic, Haukeland University Hospital, Bergen, Hordaland, Norway

The Autoimmune Regulator, AIRE, acts as a transcriptional regulator in the thymus, facilitating ectopic expression of thousands of genes important for the process of negative T-cell selection and immunological tolerance to self. Pathogenic variants in the gene encoding AIRE are causing Autoimmune polyendocrine syndrome type 1 (APS-1), defined by multiorgan autoimmunity and chronic mucocutaneous candidiasis. More recently, Genome Wide Association Studies (GWAS) have also implicated AIRE in several common organ-specific autoimmune diseases including Autoimmune primary adrenal insufficiency, type 1 diabetes and pernicious anemia. We developed a highly sensitive cell-system approach based on HEK293FT cells transfected with AIRE that allowed us to characterise and functionally evaluate the transcriptional potential of genetic variants in the AIRE gene. We confirm that our cell system recapitulates the expression of the vast majority of known AIRE induced genes including well-characterised tissue restricted antigens (TRAs), but also increases the total number of identified AIRE induced genes by an order of magnitude compared to previously published strategies. The approach differentiates between categories of AIRE variants on the transcriptional level, including the nonsense variant p.R257* (near complete loss of function), the p.C311Y variant associated with dominantly inherited APS-1 (severely impaired function), and the polygenic risk variant p.R471C (slightly increased function) linked to common organ-specific autoimmunity. The increased activity of p.R471C compared to wildtype indicates different molecular mechanisms for monogenic and polygenic AIRE related autoimmunity.