Abstract
Apoptosis is a programmed cell death process, whose complexity led researchers to build mathematical models that could help to identify its crucial steps. In previous works, we theoretically analyzed and numerically simulated a model that describes a pathway from an external stimulus to caspase-3 activation. Here, the results of experiments performed on populations of synchronized cells treated with the inducer Apo2L/TRAIL are reported and are compared with model predictions. In particular, we have compared in vitro and in silico results relevant to the time evolutions of caspase-3 and caspase-8 activities, as well as of the dead cells fractions. In addition, the effect of the BAR gene silencing was evaluated. Caspase-3 activation and cell death is faster in silenced than in nonsilenced cells, thus confirming previous simulation results. Interestingly, Apo2L/TRAIL treatment in itself reduces the BAR gene expression. The qualitative agreement between model predictions and cell cultures behavior suggests that the model captures the essential features of the biological process and could be a tool in further studies of caspases activation. In this manuscript, we report the results of in vitro experiments aimed at revealing the dynamics of caspase activation in a cell population. A qualitative agreement between these results and a mathematical model describing a pathway from an external stimulus to caspase-3 activation was obtained, thus showing that the model captures the essential features of the biological process and may be a reliable tool in further studies of caspase activation.
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Abbreviations
- Apo2L/TRAIL:
-
Apoptosis-inducing ligand 2/Tumor necrosis factor-related apoptosis-inducing ligand
- siRNA:
-
Small interfering RNA
- RT-PCR:
-
Reverse transcriptase polymerase chain reaction
- FACS:
-
Fluorescence activated cell sorter
References
Auffray C, Imbeaud S, Roux-Rouquié M, Hood L (2003) From functional genomics to system biology: concepts and practices. CR Biol 326:879–892. doi:10.1016/j.crvi.2003.09.033
Bagci EZ, Vodovotz Y, Billiar TR, Ermentrout GB, Bahar I (2006) Bistability in apoptosis: roles of Bax, bcl-2 and mitochondrial permeability transition pores. Biophys J 90:1546–1559. doi:10.1529/biophysj.105.068122
Bentele M, Lavrik I, Ulrich M, Stosser S, Heermann DH, Kalthoff H, Krammer PH, Eils R (2004) Mathematical modelling reveals threshold mechanism in CD95-induced apoptosis. J Cell Biol 166:839–851. doi:10.1083/jcb.200404158
Bialik S, Cryns VL, Drincic A, Miyata S, Wollowick AL, Srinivasan A, Kitsis RN (1999) The mitochondrial apoptotic pathway is activated by serum and glucose deprivation in cardiac myocytes. Circ Res 85:403–414
Bortner CD, Oldenburg NB, Cidlowski JA (1995) The role of DNA fragmentation in apoptosis. Trends Cell Biol 5:21–26. doi:10.1016/S0962-8924(00)88932-1
Carotenuto L, Pace V, Bellizzi D, De Benedictis G (2007a) Equilibrium, stability and dynamical response in a model of the extrinsic apoptosis pathway. J Biol Syst 15:261–285. doi:10.1142/S0218339007002192
Carotenuto L, Pace V, Bellizzi D, De Benedictis G (2007b) Dynamical Analysis of the Programmed Cell Death Pathway. In: Tzafestas S (ed) Proceedings of the European Control Conference 2007 (ECC'07), Kos, 2–5 July 2007, pp 3747–3754. http://biologia.unical.it/genetica/Carotenuto_et_al.pdf
Degterev A, Yuan J (2008) Expansion and evolution of cell death programmes. Nat Rev Mol Cell Biol 9:378–390. doi:10.1038/nrm2393
Dubrez-Daloz L, Dupoux A, Cartier J (2008) IAPs: more than just inhibitors of apoptosis proteins. Cell Cycle 7:1036–1046
Eissing T, Conzelmann H, Gilles ED, Allgower F, Bullinger E, Scheurich P (2004) Bistability analyses of a caspase activation model for receptor-induced apoptosis. J Biol Chem 279:36892–36897. doi:10.1074/jbc.M404893200
Eissing T, Allgower F, Bullinger E (2005) Robustness properties of apoptosis models with respect to parameter variations and intrinsic noise. Syst Biol 12:221–228
Fussenegger M, Bailey JE, Varner JA (2000) Mathematical model of caspase function in apoptosis. Nat Biotechnol 18:768–774. doi:10.1038/81208
Hague A, Paraskeva C (2004) Apoptosis and disease: a matter of cell fate. Cell Death Differ 11:1366–1372. doi:10.1038/sj.cdd.4401497
King MP, Attardi G (1996) Isolation of human cell lines lacking mitochondrial DNA. Methods Enzymol 264:304–313. doi:10.1016/S0076-6879(96)64029-4
Krysko DV, Vanden Berghe T, D'Herde K, Vandenabeele P (2008) Apoptosis and necrosis: detection, discrimination and phagocytosis. Methods 44:205–221. doi:10.1016/j.ymeth.2007.12.001
Legewie S, Bluthgen N, Herzel H (2006) Mathematical modelling identifies inhibitors of apoptosis as mediators of positive feedback and bistability. PLos Comput Biol 2:1061–1073. doi:10.1371/journal.pcbi.0020120
Marsden VS, Strasser A (2003) Control of apoptosis in the immune system: Bcl-2, BH3-Only proteins and more. Annu Rev Immunol 21:71–105. doi:10.1146/annurev.immunol.21.120601.141029
Opferman JY, Korsmeyer SJ (2003) Apoptosis in the development and maintenance of the immune system. Nat Immunol 4:410–415. doi:10.1038/ni0503-410
Pillich RT, Scarsella G, Risuleo G (2005) Reduction of apoptosis through the mitochondrial pathway by the administration of acetyl-l-carnitine to mouse fibroblasts in culture. Exp Cell Res 306:1–8. doi:10.1016/j.yexcr.2005.01.019
O'Connor CL, Anguissola S, Huber HJ, Dussmann H, Prehn JH, Rehm M (2008) Intracellular signaling dynamics during apoptosis execution in the presence or absence of X-linked-inhibitor-of-apoptosis-protein. Biochim Biophys Acta 1783:1903–1913. doi:10.1016/j.bbamcr.2008.05.025
Rehm M, Huber HJ, Dussmann H, Prehn JHM (2006) Systems analysis of effector caspase activation and its control by X-linked inhibitor of apoptosis protein. EMBO J 25:4338
Salvesen GS, Riedl SJ (2008) Caspase mechanisms. Adv Exp Med Biol 615:13–23
Stucki JW, Simon HU (2005) Mathematical modelling of the regulation of caspase-3 activation and degradation. J Theor Biol 23:123–131. doi:10.1016/j.jtbi.2004.11.011
Takeda S, Iwai A, Nakashima M et al (2007) LKB1 is crucial for TRAIL-mediated apoptosis induction in osteosarcoma. Anticancer Res 27:761–768
Van England M, Nieland LJW, Ramaekers FCS, Schutte B, Reutelingsperger CP (1998) Annexin V-Affinity Assay: A review on an apoptosis detection system based on phosphatydilserine exposure. Cytometry 31:1. doi:10.1002/(SICI)1097-0320(19980101)31:1<1::AID-CYTO1>3.0.CO;2-R
Wolkenhauer O (2007) Defining systems biology: an engineering perspective. IET Syst Biol 1:204–206. doi:10.1049/iet-syb:20079017
Zhang H, Xu Q, Krajewski S, Krajewska M, Xie Z, Fuess S, Kitada S, Pawlowski K, Godzik A, Reed JC (2000) BAR: An apoptosis regulator at the intersection of caspases and Bcl-2 family proteins. Proc Natl Acad Sci U S A 97:2597–2602. doi:10.1073/pnas.97.6.2597
Acknowledgments
The work was supported by the EU project “European Challenge for Healthy Ageing” (No. QLRT-2001-00128, Call Identifier QOL-2001-3; to GDB) and by Fondo Sociale Europeo (Ph.D. course in Molecular Biopathology, University of Calabria, Italy).
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Vincenza Pace and Dina Bellizzi contributed equally to this work.
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Pace, V., Bellizzi, D., Giordano, F. et al. Experimental testing of a mathematical model relevant to the extrinsic pathway of apoptosis. Cell Stress and Chaperones 15, 13–23 (2010). https://doi.org/10.1007/s12192-009-0118-9
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DOI: https://doi.org/10.1007/s12192-009-0118-9