title: Targeting neddylation induces DNA damage and checkpoint activation and sensitizes chronic lymphocytic leukemia B cells to alkylating agents.
abstract: Microenvironment-mediated upregulation of the B-cell receptor (BCR) and nuclear factor-κB (NF-κB) signaling in CLL cells resident in the lymph node and bone marrow promotes apoptosis evasion and clonal expansion. We recently reported that MLN4924 (pevonedistat), an investigational agent that inhibits the NEDD8-activating enzyme (NAE), abrogates stromal-mediated NF-κB pathway activity and CLL cell survival. However, the NAE pathway also assists degradation of multiple other substrates. MLN4924 has been shown to induce DNA damage and cell cycle arrest, but the importance of this mechanism in primary neoplastic B cells has not been studied. Here we mimicked the lymph node microenvironment using CD40 ligand (CD40L)-expressing stroma and interleukin-21 (IL-21) to find that inducing proliferation of the primary CLL cells conferred enhanced sensitivity to NAE inhibition. Treatment of the CD40-stimulated CLL cells with MLN4924 resulted in deregulation of Cdt1, a DNA replication licensing factor, and cell cycle inhibitors p21 and p27. This led to DNA damage, checkpoint activation and G2 arrest. Alkylating agents bendamustine and chlorambucil enhanced MLN4924-mediated DNA damage and apoptosis. These events were more prominent in cells stimulated with IL-21 compared with CD40L alone, indicating that, following NAE inhibition, the culture conditions were able to direct CLL cell fate from an NF-κB inhibition to a Cdt1 induction program. Our data provide insight into the biological consequences of targeting NAE in CLL and serves as further rationale for studying the clinical activity of MLN4924 in CLL, particularly in combination with alkylating agents.
authors.affiliation: Knight Cancer Institute, Oregon Health and Science University, Portland, OR, USA.
authors.forename: C
authors.initials: C
authors.lastname: Paiva
authors.affiliation: Geisel School of Medicine at Dartmouth, Hanover, NH, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: Godbersen
authors.affiliation: Millennium Pharmaceuticals, Inc., a wholly owned subsidiary of Takeda Pharmaceutical Company Ltd, Cambridge, MA, USA.
authors.forename: A
authors.initials: A
authors.lastname: Berger
authors.affiliation: Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.
authors.forename: J R
authors.initials: JR
authors.lastname: Brown
authors.affiliation: Knight Cancer Institute, Oregon Health and Science University, Portland, OR, USA.
authors.forename: A V
authors.initials: AV
authors.lastname: Danilov
mesh_terms: D000477:Alkylating Agents / Q000008:administration & dosage*; D017209:Apoptosis / Q000187:drug effects; D001402:B-Lymphocytes / Q000378:metabolism; D000069461:Bendamustine Hydrochloride / Q000008:administration & dosage; D019013:CD40 Antigens / Q000096:biosynthesis* / Q000235:genetics; D023201:CD40 Ligand / Q000235:genetics*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003517:Cyclopentanes / Q000008:administration & dosage; D004249:DNA Damage / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007378:Interleukins / Q000096:biosynthesis / Q000235:genetics; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D000075687:NEDD8 Protein; D016328:NF-kappa B / Q000235:genetics; D011743:Pyrimidines / Q000008:administration & dosage; D044764:Ubiquitin-Activating Enzymes / Q000037:antagonists & inhibitors / Q000235:genetics*; D014452:Ubiquitins / Q000235:genetics; D000098546:Interleukin-21
chemical_list: D000477:Alkylating Agents; D019013:CD40 Antigens; D003517:Cyclopentanes; D007378:Interleukins; D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; D016328:NF-kappa B; D011743:Pyrimidines; D014452:Ubiquitins; D023201:CD40 Ligand; D000069461:Bendamustine Hydrochloride; D044764:Ubiquitin-Activating Enzymes; C491802:NAE protein, human; D000098546:Interleukin-21; C539933:pevonedistat