LINCSapplication/xmlhttp://lincsportal.ccs.miami.edu/dcic/api/download?path=LINCS_Data/HMS_LINCS&file=LDS-1222.tar.gzprimary200OKUnknownJeremie Roux, Marc Hafner, Samuel BandaraNIH 1U54HG006097-01; NIH 1U54HL127365-01Fluorescence imagingFluorescence intensityTNF10http://lincsportal.ccs.miami.edu/datasets/#/view/LDS-1222HMS LINCS (Harvard Medical School)This analysis dataset presents calculated parameters fit to a phenomenological model that defines an initiator caspase threshold that underlies cell-to-cell variability in cell killing by TRAIL (TNF10) and therapeutic antibody TRAIL receptor agonists. The analysis is based on live-cell, microscopy-based fate tracking of cells following perturbation with TRAIL, therapeutic antibody TRAIL receptor agonists, and other agents that modulate the cellular fate decision.LINCSWhen cells are exposed to death ligands such as TRAIL, a fraction undergoes apoptosis and a fraction survives; if surviving cells are re-exposed to TRAIL, fractional killing is once again observed. Therapeutic antibodies directed against TRAIL receptors also cause fractional killing, even at saturating concentrations, limiting their effectiveness. Fractional killing arises from cell-to-cell fluctuations in protein levels (extrinsic noise), but how this results in a clean bifurcation between life and death remains unclear. In this paper, we identify a threshold in the rate and timing of initiator caspase activation that distinguishes cells that live from those that die; by mapping this threshold, we can predict fractional killing of cells exposed to natural and synthetic agonists alone or in combination with sensitizing drugs such as bortezomib. A phenomenological model of the threshold also quantifies the contributions of two resistance genes (c-FLIP and Bcl-2), providing new insight into the control of cell fate by opposing pro-death and pro-survival proteins and suggesting new criteria for evaluating the efficacy of therapeutic TRAIL receptor agonists.Fractional killing arises from cell-to-cell variability in overcoming a caspase activity threshold.Roux Jérémie J, Hafner Marc M, Bandara Samuel S, Sims Joshua J JJ, Hudson Hannah H, Chai Diana D, Sorger Peter K PKfalseLive-cell imaging of cell-to-cell variability in caspase-8 FRET reporter activity and cell fate trajectory modeling-2016-03-222016-03-22LDS-122225953765