Most cancer deaths result from progression of therapy resistant disease, yet our understanding of this phenotype is limited. Cancer therapies generate stress signals that act upon mitochondria to initiate apoptosis. Mitochondria isolated from neuroblastoma cells were exposed to tBid or Bim, death effectors activated by therapeutic stress. Multidrug-resistant tumor cells obtained from children at relapse had markedly attenuated Bak and Bax oligomerization and cytochrome c release (surrogates for apoptotic commitment) in comparison with patient-matched tumor cells obtained at diagnosis. Electron microscopy identified reduced ER-mitochondria-associated membranes (MAMs; ER-mitochondria contacts, ERMCs) in therapy-resistant cells, and genetically or biochemically reducing MAMs in therapy-sensitive tumors phenocopied resistance. MAMs serve as platforms to transfer Ca2+ and bioactive lipids to mitochondria. Reduced Ca2+ transfer was found in some but not all resistant cells, and inhibiting transfer did not attenuate apoptotic signaling. In contrast, reduced ceramide synthesis and transfer was common to resistant cells and its inhibition induced stress resistance. We identify ER-mitochondria-associated membranes as physiologic regulators of apoptosis via ceramide transfer and uncover a previously unrecognized mechanism for cancer multidrug resistance.

Cancer Biology Program Perelman School of Medicine University of Pennsylvania Philadelphia PA USA

Department of Biostatistics Perelman School of Medicine University of Pennsylvania Philadelphia PA USA

Department of Experimental Biology Faculty of Science Masaryk University Brno Czech Republic

Department of Medicine Perelman School of Medicine University of Pennsylvania Philadelphia PA USA

Department of Neurology Columbia University Irving Medical Center New York NY USA

Department of Pediatrics Emory University School of Medicine Atlanta GA USA

Department of Pediatrics Perelman School of Medicine University of Pennsylvania Philadelphia PA USA

Department of Radiation Oncology Perelman School of Medicine University of Pennsylvania Philadelphia PA USA

Division of Oncology and Center for Childhood Cancer Research The Children's Hospital of Philadelphia Philadelphia PA USA

International Clinical Research Center St Anne's University Hospital Brno Czech Republic

MitoCare Center Department of Pathology Anatomy and Cell Biology Thomas Jefferson University Philadelphia PA USA

Mitochondrial Medicine Frontier Program Division of Human Genetics The Children's Hospital of Philadelphia Philadelphia PA USA

TTUHSC Cancer Center Texas Tech University Health Sciences Center Lubbock TX USA

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Mitoribosomal synthetic lethality overcomes multidrug resistance in MYC-driven neuroblastoma