Session: 651. Multiple Myeloma and Plasma Cell Dyscrasias: Basic and Translational: Poster I
Hematology Disease Topics & Pathways:
Plasma Cell Disorders, Diseases, Metabolism, Lymphoid Malignancies, Biological Processes
Materials and methods. MM cell lines harboring CD38 positive and negative fractions (KMS-12BM, KMS-11) were sorted according to CD38 expression. Intracellular NAD+ and NADH concentrations between CD38 positive and negative cells were analyzed using NAD/NADH Assay kit. CD38 positive and negative MM cells were subjected to metabolome and proteome analysis using Shimadzu TQ8050 GC–MS/MS and TripleTOF 5600 respectively. Metabolites and proteins significantly enriched in CD38 negative MM cells were analyzed using MetaboAnalyst and Metascape. Cell cycle status between CD38 positive and negative cells were determined by flow cytometry after staining by BRDU and 7AAD. CD38 positive MM cell lines (NCI-H929 and KMS-12PE) and patient derived bone marrow cells were treated with 78c, a CD38 NADase inhibitor, in vitro. MM cell viability were determined by flow cytometry post Annexin V and PI staining. Differences in metabolites between 78c treated and control MM cell lines were also analyzed using Shimadzu TQ8050 GC–MS/MS.
Results. Higher NAD+ and NAD+/NADH ratio was observed in CD38 negative fraction of MM cell lines compared to their CD38 positive counterparts, demonstrating that cell surface CD38 expression influences intracellular NAD+ concentration. Both metabolome and proteome analysis revealed that CD38 negative cells tend to have higher glycolytic activity compared to CD38 positive cells. Significant suppression of cell cycle, accompanying G0/G1 phase arrest, was observed in CD38 negative MM cells, indicating that metabolic shift in CD38 negative MM cells may lead to change in cell proliferation. Marked increase of NAD+/NADH ratio was observed in 78c treated MM cell lines compared to control, proving that CD38 NADase inhibiton indeed affects intracellular NAD+ concentration in MM cells. 78c was capable of inducing cell death in MM cell lines and patient derived MM cells, accompanying cell cycle arrest. Metabolites significantly upregulated in 78c treated MM cells compared to control were associated with glycolysis, demonstrating that CD38 NADase activity has a significant effect on MM cell metabolism.
Conclusions. CD38 is the major NADase in mammalian tissues, involved in catabolism of NAD+. Although CD38 is highly expressed in normal plasma cells and MM cells, its role in MM cell biology has not been studied in detail. By comparing CD38 positive and negative cells and using CD38 NADase inhibitor, we showed for the first time that CD38 on MM cells decrease intracellular NAD+, reduces intracellular glycolysis and as a result, has an influence on cell cycle. The present study sheds light on the significance of CD38 enzyme activity in MM cell biology and may also contribute to understanding the mechanism of resistance to CD38 targeted therapy.
Disclosures: Kawano: Ono pharmaceutical: Honoraria; Takeda Pharmaceuticals: Honoraria; Bristol Myers Squibb: Honoraria; Sanofi: Honoraria; Janssen Pharmaceuticals: Honoraria.
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