title: Temporal Heterogeneity Metrics in Apoptosis Induced by Anticancer Drugs.
abstract: The apoptotic process is highly heterogeneous and asynchronous. A long-standing question is how many parameters define the time and reversibility of the apoptotic response at a single-cell level. We characterized at the single-cell and population levels the time sequence of apoptotic events in response to anti-cancer drugs using extrinsic and intrinsic apoptotic stimuli. We show that the temporal sequence of major apoptotic events is the same in response to all anti-cancer drugs studied: the apoptotic volume decrease and Na+ influx occur rapidly and are tightly coordinated with mitochondrial outer membrane depolarization (MOMP), mitochondrial inner membrane depolarization and a decrease in the production of reactive oxygen species (ROS). Phosphatidylserine externalization usually starts after MOMP and precedes caspase 3/7 activation. Activation of caspases 3/7 is a slow process that always starts after MOMP, with significant delay. Cell-to-cell variability of the MOMP onset is described by Gaussian distribution, whereas the γ-distribution model describes cellular variability in the duration of MOMP-to-caspase activation stages. Cells from the pre-MOMP stage to the after-caspase 3/7 activation stage coexist for many hours. We demonstrated by FACS that cells in the pre-MOMP stage can recover after apoptotic stimuli, rarely recover after MOMP but before caspase 3/7 activation, and are unable to recover after caspase 3/7 activation. We propose a double-stroke model for apoptosis execution.
authors.affiliation: A.N. Belozersky Institute of Physico-Chemical Biology, Department of Cell Biology and Histology, Faculty of Biology, M.V. Lomonosov Moscow State University, Moscow, Russia (IV)
authors.forename: Ivan
authors.initials: I
authors.lastname: Vorobjev
authors.affiliation: Cellular and Molecular Medicine Program, Boston Children's Hospital, Department of Pediatrics, Harvard Medical School, Boston, Massachusetts (NSB).
authors.forename: Natasha S
authors.initials: NS
authors.lastname: Barteneva
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D053142:Mitochondrial Size / Q000187:drug effects; D008954:Models, Biological; D010718:Phosphatidylserines / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D059010:Single-Cell Analysis; D013997:Time Factors
chemical_list: D000970:Antineoplastic Agents; D010718:Phosphatidylserines; D017382:Reactive Oxygen Species; D053148:Caspase 3; D053179:Caspase 7
keywords: MOMP; TMRE; TRAIL; actinomycin D; apoptosis; caspase; cell heterogeneity; cell volume; etoposide; staurosporine
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