Immunoglobulin light chain (AL) amyloidosis is caused by a small, minimally proliferating B-cell/plasma-cell clone secreting a patient-unique, aggregation-prone, toxic light chain (LC). The pathogenicity of LCs is encrypted in their sequence, yet molecular determinants of amyloidogenesis are poorly understood. Higher rates of N-glycosylation among clonal κ LCs from patients with AL amyloidosis compared to other monoclonal gammopathies indicate that this post-translational modification is associated with a higher risk of developing AL amyloidosis. Here, we exploited LC sequence information from previously published amyloidogenic and control clonal LCs and from a series of 220 patients with AL amyloidosis or multiple myeloma followed at our Institutions to define sequence and spatial features of N-glycosylation, combining bioinformatics, biochemical, proteomics, structural and genetic analyses. We found peculiar sequence and spatial pattern of N-glycosylation in amyloidogenic κ LCs, with most of the N-glycosylation sites laying in the framework region 3, particularly within the E strand, and consisting mainly of the NFT sequon, setting them apart with respect to non-amyloidogenic clonal LCs. Our data further support a potential role of N-glycosylation in determining the pathogenic behavior of a subset of amyloidogenic LCs and may help refine current N-glycosylation-based prognostic assessments for patients with monoclonal gammopathies.
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The data generated in this study are available within the article and its Supplementary Data Files. LC sequences have been deposited to GenBank (MZ595009-MZ595094, OM885091-OM885224).
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We thank Andrea Patrignani and staff of the Functional Genomic Center Zurich for PacBio library preparation and sequencing. This work was supported by grants from the Amyloidosis Foundation (MNu), the Italian Ministry of Health (Ricerca Finalizzata, grant #GR-2018-12368387) (MNu), the Italian Ministry of Research and Education (PRIN 20207XLJB2) (SR, GP), the CARIPLO Foundation (grant #2018-0257) (MNu), Fondazione ARISLA (project TDP-43-STRUCT) (SR), Cancer Research UK [C355/A26819], FC AECC and AIRC under the Accelerator Award Program (BP, GM, GP, MNu). We would like to acknowledge the use of the Boston University AL-Base, supported by HL68705, in this work.
PC, GP, and MNu are inventors on a patent application related to immunoglobulin sequencing.
Ethics approval and consent to participate
Clinical records and biological samples were from subjects referred to the Italian Amyloid Center or to the Department of Hematology, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy and to the University Hospital of Navarra, Pamplona, Spain for a diagnostic workout in the suspicion of systemic AL amyloidosis or MM. Per the Declaration of Helsinki, all patients gave their written informed consent for the use of their clinical data and biological samples for research purposes, and this study was approved by the local Institutional Review Board.
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Nevone, A., Girelli, M., Mangiacavalli, S. et al. An N-glycosylation hotspot in immunoglobulin κ light chains is associated with AL amyloidosis. Leukemia (2022). https://doi.org/10.1038/s41375-022-01599-w