title: Nedd8-activating enzyme inhibitor MLN4924 provides synergy with mitomycin C through interactions with ATR, BRCA1/BRCA2, and chromatin dynamics pathways.
abstract: MLN4924 is an investigational small-molecule inhibitor of the Nedd8-activating enzyme currently in phase I clinical trials. MLN4924 induces DNA damage via rereplication in most cell lines. This distinct mechanism of DNA damage may affect its ability to combine with standard-of-care agents and may affect the clinical development of MLN4924. As such, we studied its interaction with other DNA-damaging agents. Mitomycin C, cisplatin, cytarabine, UV radiation, SN-38, and gemcitabine demonstrated synergy in combination with MLN4924 in vitro. The combination of mitomycin C and MLN4924 was shown to be synergistic in a mouse xenograft model. Importantly, depletion of genes within the ataxia telangiectasia and Rad3 related (ATR) and BRCA1/BRCA2 pathways, chromatin modification, and transcription-coupled repair reduced the synergy between mitomycin C and MLN4924. In addition, comet assay demonstrated increased DNA strand breaks with the combination of MLN4924 and mitomycin C. Our data suggest that mitomycin C causes stalled replication forks, which when combined with rereplication induced by MLN4924 results in frequent replication fork collisions, leading to cell death. This study provides a straightforward approach to understand the mechanism of synergy, which may provide useful information for the clinical development of these combinations.
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Khristofer
authors.initials: K
authors.lastname: Garcia
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Blank
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: David C
authors.initials: DC
authors.lastname: Bouck
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Xiaozhen J
authors.initials: XJ
authors.lastname: Liu
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Darshan S
authors.initials: DS
authors.lastname: Sappal
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Greg
authors.initials: G
authors.lastname: Hather
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Katherine
authors.initials: K
authors.lastname: Cosmopoulos
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Thomas
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Mike
authors.initials: M
authors.lastname: Kuranda
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Pickard
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Ray
authors.initials: R
authors.lastname: Liu
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Syamala
authors.initials: S
authors.lastname: Bandi
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts.
authors.forename: Peter G
authors.initials: PG
authors.lastname: Smith
authors.affiliation: Authors' Affiliations: Departments of Discovery, Clinical Biostatistics, and Information Technology, Takeda Pharmaceuticals International Co., Cambridge, Massachusetts eric.lightcap@takeda.com.
authors.forename: Eric S
authors.initials: ES
authors.lastname: Lightcap
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D064007:Ataxia Telangiectasia Mutated Proteins / Q000235:genetics / Q000378:metabolism; D019313:BRCA1 Protein / Q000235:genetics* / Q000378:metabolism; D024682:BRCA2 Protein / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D002843:Chromatin / Q000187:drug effects / Q000235:genetics; D003517:Cyclopentanes / Q000008:administration & dosage*; D004249:DNA Damage / Q000187:drug effects; D004357:Drug Synergism; D006801:Humans; D051379:Mice; D016685:Mitomycin / Q000008:administration & dosage*; D011743:Pyrimidines / Q000008:administration & dosage*; D044764:Ubiquitin-Activating Enzymes / Q000037:antagonists & inhibitors* / Q000235:genetics; D014466:Ultraviolet Rays; D023041:Xenograft Model Antitumor Assays
chemical_list: D019313:BRCA1 Protein; D024682:BRCA2 Protein; C551750:BRCA2 protein, human; D002843:Chromatin; D003517:Cyclopentanes; D011743:Pyrimidines; D016685:Mitomycin; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D044764:Ubiquitin-Activating Enzymes; C491802:NAE protein, human; C539933:pevonedistat