title: Mitochondrial Bcl-2 family dynamics define therapy response and resistance in neuroblastoma.
abstract: Neuroblastoma is a childhood tumor in which transient therapeutic responses are typically followed by recurrence with lethal chemoresistant disease. In this study, we characterized the apoptotic responses in diverse neuroblastomas using an unbiased mitochondrial functional assay. We defined the apoptotic set point of neuroblastomas using responses to distinct BH3 death domains providing a BH3 response profile and directly confirmed survival dependencies. We found that viable neuroblastoma cells and primary tumors are primed for death with tonic sequestration of Bim, a direct activator of apoptosis, by either Bcl-2 or Mcl-1, providing a survival dependency that predicts the activity of Bcl-2 antagonists. The Bcl-2/Bcl-xL/Bcl-w inhibitor ABT-737 showed single-agent activity against only Bim:Bcl-2 primed tumor xenografts. Durable complete regressions were achieved in combination with noncurative chemotherapy even for highest risk molecular subtypes with MYCN amplification and activating ALK mutations. Furthermore, the use of unique isogenic cell lines from patients at diagnosis and at the time of relapse showed that therapy resistance was not mediated by upregulation of Bcl-2 homologues or loss of Bim priming, but by repressed Bak/Bax activation. Together, our findings provide a classification system that identifies tumors with clinical responses to Bcl-2 antagonists, defines Mcl-1 as the principal mediator of Bcl-2 antagonist resistance at diagnosis, and isolates the therapy resistant phenotype to the mitochondria.
authors.affiliation: Division of Hematology/Oncology, Aflac Children's Cancer Center, Children's Healthcare of Atlanta, Atlanta, GA 30322, USA. kgoldsm@emory.edu
authors.forename: Kelly C
authors.initials: KC
authors.lastname: Goldsmith
authors.forename: Michelle
authors.initials: M
authors.lastname: Gross
authors.forename: Susan
authors.initials: S
authors.lastname: Peirce
authors.forename: Dema
authors.initials: D
authors.lastname: Luyindula
authors.forename: Xueyuan
authors.initials: X
authors.lastname: Liu
authors.forename: Annette
authors.initials: A
authors.lastname: Vu
authors.forename: Michael
authors.initials: M
authors.lastname: Sliozberg
authors.forename: Rong
authors.initials: R
authors.lastname: Guo
authors.forename: Huaqing
authors.initials: H
authors.lastname: Zhao
authors.forename: C Patrick
authors.initials: CP
authors.lastname: Reynolds
authors.forename: Michael D
authors.initials: MD
authors.lastname: Hogarty
mesh_terms: D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D008928:Mitochondria / Q000378:metabolism*; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D009368:Neoplasm Transplantation; D009447:Neuroblastoma / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000502:physiology
chemical_list: D051017:Apoptosis Regulatory Proteins; C118024:BLID protein, human; C579094:Mcl1 protein, mouse; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D019253:Proto-Oncogene Proteins c-bcl-2