Robin Wijngaard, Caspar I. van der Made, Sema Kalkan Uçar, Gayatri Ramakrishnan, Man Wang, Johannes Brand, Jill A. Rosenfeld, Tiphanie P. Vogel, Sarah K. Nicholas, Monika Weisz‐Hubshman, Clara D.M. van Karnebeek, Eric J. Allenspach, Taylor E. Gardiner, Sumudu Perera Kimmantudawage, Zornitza Stark, Ruth K Armstrong, Janine Campbell, Stefano Volpi, Enrico Drago, Marco Gattorno, Alice Grossi, Isabella Ceccherini, Alfredo Cabrera‐Orefice, Bente Siebels, Thomas Mair, Hartmut Schlüter, Ruben L. Smeets, Ronald van Beek, Ingrid Goebel, Katrin Küchler, Søren W. Gersting, Alexander Hoischen, Lisenka E.L.M. Vissers, Ron A. Wevers, Catherine Meyer-Schwesinger, Saskia B. Wortmann, M S Oud, Sergio Guerrero‐Castillo
Monoallelic variants in catalytic immunoproteasome subunits have recently been linked to proteasome-associated autoinflammatory syndromes with immunodeficiency (PRAAS-ID), yet their molecular mechanisms and clinical spectra are not fully defined. In this study, seven individuals from five unrelated families carrying five distinct monoallelic PSMB8 variants were identified. Individuals presented with neonatal-onset immunodeficiency characterized by recurrent infections, B cell lymphopenia, and hypogammaglobulinemia requiring immunoglobulin replacement. Inflammatory manifestations of variable severity included enteropathy, hepatitis, myositis, and inflammatory lung disease. Additional findings included leukocyte vacuolization in blood and bone marrow. Pathogenic variants in immunoproteasome subunits were analyzed to identify structural features associated with dominant-negative behavior. Immunoproteasome assembly and activity were investigated using complexome profiling, immunoblotting, and in-gel activity assays in proband-derived fibroblasts and transfected HEK293T cells, with downstream effects assessed by proteomic and RT-qPCR analyses. Mutant PSMB8 subunits were inefficiently incorporated into immunoproteasome complexes, leading to impaired assembly, including reduced fully assembled complexes and accumulation of assembly intermediates. This defect was accompanied by activation of the integrated stress response alongside impaired immune signaling. Monoallelic pathogenic variants in PSMB8, PSMB9, and PSMB10 associated with PRAAS-ID affected residues that are highly conserved and biophysically similar between the three immunoproteasome catalytic subunits. These shared structural features may help identify additional variants with similar disruptive effects on immunoproteasome assembly. Together, our data show that monoallelic PSMB8 variants disrupt immunoproteasome assembly, resulting in clinically variable disease with immunodeficiency and systemic inflammation. Our findings support immunoproteasome assembly disruption as a unifying dominant-negative mechanism underlying PRAAS-ID.