title: Mito-priming as a method to engineer Bcl-2 addiction.
abstract: Most apoptotic stimuli require mitochondrial outer membrane permeabilization (MOMP) in order to execute cell death. As such, MOMP is subject to tight control by Bcl-2 family proteins. We have developed a powerful new technique to investigate Bcl-2-mediated regulation of MOMP. This method, called mito-priming, uses co-expression of pro- and anti-apoptotic Bcl-2 proteins to engineer Bcl-2 addiction. On addition of Bcl-2 targeting BH3 mimetics, mito-primed cells undergo apoptosis in a rapid and synchronous manner. Using this method we have comprehensively surveyed the efficacy of BH3 mimetic compounds, identifying potent and specific MCL-1 inhibitors. Furthermore, by combining different pro- and anti-apoptotic Bcl-2 pairings together with CRISPR/Cas9-based genome editing, we find that tBID and PUMA can preferentially kill in a BAK-dependent manner. In summary, mito-priming represents a facile and robust means to trigger mitochondrial apoptosis.
authors.affiliation: Cancer Research UK Beatson Institute, Switchback Road, Glasgow G61 1BD, UK.|Institute of Cancer Sciences, University of Glasgow, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Jonathan
authors.identifier: http://orcid.org/0000-0003-3768-2378
authors.initials: J
authors.lastname: Lopez
authors.affiliation: Cancer Research UK Beatson Institute, Switchback Road, Glasgow G61 1BD, UK.|Institute of Cancer Sciences, University of Glasgow, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Margaux
authors.initials: M
authors.lastname: Bessou
authors.affiliation: Cancer Research UK Beatson Institute, Switchback Road, Glasgow G61 1BD, UK.|Institute of Cancer Sciences, University of Glasgow, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Joel S
authors.initials: JS
authors.lastname: Riley
authors.affiliation: Cancer Research UK Beatson Institute, Switchback Road, Glasgow G61 1BD, UK.|Institute of Cancer Sciences, University of Glasgow, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Evangelos
authors.initials: E
authors.lastname: Giampazolias
authors.affiliation: Institute for Biochemistry and Molecular Biology, ZBMZ, Faculty of Medicine, University of Freiburg, 79104 Freiburg, Germany.
authors.forename: Franziska
authors.initials: F
authors.lastname: Todt
authors.affiliation: Cancer Research UK Beatson Institute, Switchback Road, Glasgow G61 1BD, UK.|Institute of Cancer Sciences, University of Glasgow, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Tony
authors.initials: T
authors.lastname: Rochegüe
authors.affiliation: Department of Immunology, University of Washington, Campus Box 358059, 750 Republican Street, Seattle, Washington, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Oberst
authors.affiliation: Department of Immunology, St Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, Tennessee 38105, USA.
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Green
authors.affiliation: Institute for Biochemistry and Molecular Biology, ZBMZ, Faculty of Medicine, University of Freiburg, 79104 Freiburg, Germany.|BIOSS, Centre for Biological Signaling Studies, University of Freiburg, 79104 Freiburg, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Edlich
authors.affiliation: Cancer Research UK Beatson Institute, Switchback Road, Glasgow G61 1BD, UK.|Institute of Cancer Sciences, University of Glasgow, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Ichim
authors.affiliation: Cancer Research UK Beatson Institute, Switchback Road, Glasgow G61 1BD, UK.|Institute of Cancer Sciences, University of Glasgow, Garscube Estate, Switchback Road, Glasgow G61 1BD, UK.
authors.forename: Stephen W G
authors.initials: SW
authors.lastname: Tait
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D064113:CRISPR-Cas Systems; D002460:Cell Line; D005786:Gene Expression Regulation; D005818:Genetic Engineering; D006801:Humans; D051379:Mice; D051336:Mitochondrial Membranes / Q000502:physiology*; D010446:Peptide Fragments / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism*
chemical_list: D051017:Apoptosis Regulatory Proteins; C474007:Bax protein (53-86); D010446:Peptide Fragments; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2
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