L-Asparaginase treatment induces reversible immunoregulatory and immunosuppressive effects in non-malignant B cells in a model of T-cell dependent B cell activation

Hadzic, A.; Garcia-Marquez, M. A.; Bannasch, P.; Haindl, N.; Frey, K.; Kirmaier, M.; Funk, M. A.; Tischmacher, A.; Tu, J.; Carboniero, A.; Vona, L.; Oganesian, S.; Cordas dos Santos, D. M.; Schloesser, H.; von Bergwelt-Baildon, M.; Schmitz, W.; Theurich, S.

doi:10.1101/2024.11.27.625617

bioRxiv · 10.1101/2024.11.27.625617

L-Asparaginase treatment induces reversible immunoregulatory and immunosuppressive effects in non-malignant B cells in a model of T-cell dependent B cell activation

Hadzic, A.·Garcia-Marquez, M. A.·Bannasch, P.·Haindl, N.·Frey, K.·Kirmaier, M.·Funk, M. A.·Tischmacher, A.·Tu, J.·Carboniero, A.·Vona, L.·Oganesian, S.·Cordas dos Santos, D. M.·Schloesser, H.·von Bergwelt-Baildon, M.·Schmitz, W.·Theurich, S.

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Abstract

Metabolic reprogramming is critical for immune cell adaptation upon activation to exert full functionality with amino acids being a key metabolic factor. While L-asparagine is a non- essential amino acid it turns out to be conditionally essential in malignant B cells due to defective asparagine synthetase making L-asparaginase a commonly used chemotherapeutic agent. Off-target enzymatic activity including glutaminolysis impacting crucial immune function. However, its effects on healthy B cells remain unclear, therefore in this study, we explored how L-asparaginase modulates the biology and function of CD40-activated B cells, using an in-vitro model. B cells from healthy donors were treated with increasing L- asparaginase concentrations and analyzed for proliferation, immune phenotype, and metabolic changes. Results showed L-asparaginase reduced B cell proliferation and homotypic clustering without inducing apoptosis, instead impairing metabolic pathways, lowering glycolysis and oxidative phosphorylation, and reducing surface markers associated with antigen-presenting cell (APC) function. Functional assays confirmed that L-asparaginase- treated B cells had diminished ability to activate T cells. Supplementing with asparagine or glutamine restored B cell proliferation and function, with glutamine slightly more effective than asparagine. Interestingly, L-asparaginase induced a regulatory B cell phenotype, marked by CD24+CD38+CD27+ expression and increased interleukin-10 and TGF-beta, suggesting a potential immunosuppressive mechanism. These findings indicate that L-asparaginase not only affects malignant cells but also impacts the function of non-malignant B cells, proposing potential therapeutic applications in B cell-driven autoimmune disorders. Further studies are needed to explore its effects at lower, clinically relevant concentrations. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=132 SRC="FIGDIR/small/625617v1_ufig1.gif" ALT="Figure 1"> View larger version (19K): org.highwire.dtl.DTLVardef@1ae8d10org.highwire.dtl.DTLVardef@af405borg.highwire.dtl.DTLVardef@1aa4c0eorg.highwire.dtl.DTLVardef@16750ea_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Hadzic, A., Garcia-Marquez, M. A., Bannasch, P., Haindl, N., Frey, K., Kirmaier, M., Funk, M. A., Tischmacher, A., Tu, J., Carboniero, A., Vona, L., Oganesian, S., Cordas dos Santos, D. M., Schloesser, H., von Bergwelt-Baildon, M., Schmitz, W., Theurich, S.. 2024-12-02. L-Asparaginase treatment induces reversible immunoregulatory and immunosuppressive effects in non-malignant B cells in a model of T-cell dependent B cell activation. https://doi.org/10.1101/2024.11.27.625617

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