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An Experimental Framework to Study the Dynamics of Autophagic Response

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Dynamics of Cell Fate Decision Mediated by the Interplay of Autophagy and Apoptosis in Cancer Cells

Part of the book series: Springer Theses ((Springer Theses))

Abstract

In this chapter we present an experimental quantitative framework for measuring kinetic parameters such as autophagy flux, time course of autophagic response, and stress/response dynamics in single cancer cells including endocrine-resistant breast cancer cells. Some primary data are presented in this chapter.

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References

  1. Abràmoff, M.D., Magalhães, P.J., Ram, S.J.: Image processing with imagej. Biophoton. Int. 11(7), 36–42 (2004)

    Google Scholar 

  2. Alon, U.: Network motifs: theory and experimental approaches. Nat. Rev. Genet. 8(6), 450–461 (2007)

    Article  Google Scholar 

  3. Ashford, T.P., Porter, K.R.: Cytoplasmic components in hepatic cell lysosomes. J. Cell Biol. 12(1), 198–202 (1962)

    Article  Google Scholar 

  4. Bampton, E.T.W., Goemans, C.G., Niranjan, D., Mizushima, N., Tolkovsky, A.M.: The dynamics of autophagy visualised in live cells: from autophagosome formation to fusion with endo/lysosomes. Autophagy 1(1), 23–36 (2005)

    Article  Google Scholar 

  5. Biggar, S.R., Crabtree, G.R.: Cell signaling can direct either binary or graded transcriptional responses. EMBO J. 20(12), 3167–3176 (2001)

    Article  Google Scholar 

  6. Brünner, N., Frandsen, T.L., Holst-Hansen, C., Bei, M., Thompson, E.W., Wakeling, A.E., Lippman, M.E., Clarke, R.: MCF7/LCC2: a 4-hydroxytamoxifen resistant human breast cancer variant that retains sensitivity to the steroidal antiestrogen ICI 182,780. Cancer Res. 53(14), 3229–3232 (1993)

    Google Scholar 

  7. Brünner, N., Boysen, B., Jirus, S., Skaar, T.C., Holst-Hansen, C., Lippman, J., Frandsen, T., Spang-Thomsen, M., Fuqua, S.A.W., Clarke, R.: Mcf7/lcc9: an antiestrogen-resistant mcf-7 variant in which acquired resistance to the steroidal antiestrogen ici 182,780 confers an early cross-resistance to the nonsteroidal antiestrogen tamoxifen. Cancer Res. 57(16), 3486–3493 (1997)

    Google Scholar 

  8. Chen, Y., Klionsky, D.J.: The regulation of autophagy–unanswered questions. J. Cell Sci. 124(2), 161–170 (2011)

    Article  Google Scholar 

  9. Cheung, W.L., Ajiro, K., Samejima, K., Kloc, M., Cheung, P., Mizzen, C.A., Beeser, A., Etkin, L.D., Chernoff, J., Earnshaw, W.C., et al.: Apoptotic phosphorylation of histone h2b is mediated by mammalian sterile twenty kinase. Cell 113(4), 507–517 (2003)

    Article  Google Scholar 

  10. Clark, S.L.: Cellular differentiation in the kidneys of newborn mice studied with the electron microscope. J. Biophys. Biochem. Cytol. 3(3), 349–362 (1957)

    Article  Google Scholar 

  11. Clarke, R., Liu, M.C., Bouker, K.B., Gu, Z., Lee, R.Y., Zhu, Y., Skaar, T.C., Gomez, B., O’Brien, K., Wang, Y., et al.: Antiestrogen resistance in breast cancer and the role of estrogen receptor signaling. Oncogene 22(47), 7316–7339 (2003)

    Article  Google Scholar 

  12. Clarke, R., Shajahan, A.N., Riggins, R.B., Cho, Y., Crawford, A., Xuan, J., Wang, Y., Zwart, A., Nehra, R., Liu, M.C.: Gene network signaling in hormone responsiveness modifies apoptosis and autophagy in breast cancer cells. J. Steroid Biochem. Mol. Biol. 114(1), 8–20 (2009)

    Article  Google Scholar 

  13. Deter, R.L., Baudhuin, P., De Duve, C.: Participation of lysosomes in cellular autophagy induced in rat liver by glucagon. J. Cell Biol. 35(2), C11–C16 (1967)

    Article  Google Scholar 

  14. Eskelinen, E.-L., Prescott, A.R., Cooper, J., Brachmann, S.M., Wang, L., Tang, X., Backer, J.M., Lucocq, J.M.: Inhibition of autophagy in mitotic animal cells. Traffic 3(12), 878–893 (2002)

    Article  Google Scholar 

  15. Eskelinen, E.-L., Reggiori, F., Baba, M., Kovács, A.L., Seglen, P.O.: Seeing is believing: the impact of electron microscopy on autophagy research. Autophagy 7(9), 935–956 (2011)

    Article  Google Scholar 

  16. Ferrell, J.E., Machleder, E.M.: The biochemical basis of an all-or-none cell fate switch in xenopus oocytes. Science 280(5365), 895–898 (1998)

    Article  Google Scholar 

  17. Filippi-Chiela, E.C., Villodre, E.S., Zamin, L.L., Lenz, G.: Autophagy interplay with apoptosis and cell cycle regulation in the growth inhibiting effect of resveratrol in glioma cells. PLoS One 6(6), e20849 (2011)

    Article  Google Scholar 

  18. Furuya, T., Kim, M., Lipinski, M., Li, J., Kim, D., Lu, T., Shen, Y., Rameh, L., Yankner, B., Tsai, L.-H., et al.: Negative regulation of VPS34 by CDK mediated phosphorylation. Mol. Cell 38(4), 500–511 (2010)

    Article  Google Scholar 

  19. Goldstein, J.C., Kluck, R.M., Green, D.R.: A single cell analysis of apoptosis: ordering the apoptotic phenotype. Ann. N. Y. Acad. Sci. 926(1), 132–141 (2000)

    Article  Google Scholar 

  20. Goldstein, J.C., Waterhouse, N.J., Juin, P., Evan, G.I., Green, D.R.: The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant. Nat. Cell Biol. 2(3), 156–162 (2000)

    Article  Google Scholar 

  21. Goldstein, J.C., Munoz-Pinedo, C., Ricci, J.E., Adams, S.R., Kelekar, A., Schuler, M., Tsien, R.Y., Green, D.R.: Cytochrome c is released in a single step during apoptosis. Cell Death Differ. 12(5), 453–462 (2005)

    Article  Google Scholar 

  22. Haley, E.M.: Autophagy induction in the quiescent state: in vitro and in vivo characterization of cellular self-cannibalism in quiescent cells. Ph.D. thesis, Princeton University (2011)

    Google Scholar 

  23. Harhaji-Trajkovic, L., Vilimanovich, U., Kravic-Stevovic, T., Bumbasirevic, V., Trajkovic, V.: AMPK-mediated autophagy inhibits apoptosis in cisplatin-treated tumour cells. J. Cell. Mol. Med. 13(9b), 3644–3654 (2009)

    Article  Google Scholar 

  24. Heatwole, V.M.: Tunel assay for apoptotic cells. In: Immunocytochemical Methods and Protocols, pp. 141–148. Springer, New York (1999)

    Google Scholar 

  25. Janji, B., Viry, E., Baginska, J., Van Moer, K., Berchem, G.: Role of autophagy in cancer and tumor progression. Autophagy: A double-edged sword-Cell survival or death, 189–215 (2013)

    Google Scholar 

  26. Kafri, R., Levy, J., Ginzberg, M.B., Oh, S., Lahav, G., Kirschner, M.W.: Dynamics extracted from fixed cells reveal feedback linking cell growth to cell cycle. Nature 494(7438), 480–483 (2013)

    Article  Google Scholar 

  27. Kaminskyy, V., Abdi, A., Zhivotovsky, B.: A quantitative assay for the monitoring of autophagosome accumulation in different phases of the cell cycle. Autophagy 7(1), 83–90 (2011)

    Article  Google Scholar 

  28. Katayama, H., Kogure, T., Mizushima, N., Yoshimori, T., Miyawaki, A.: A sensitive and quantitative technique for detecting autophagic events based on lysosomal delivery. Chem. Biol. 18(8), 1042–1052 (2011)

    Article  Google Scholar 

  29. Klionsky, D.J., Cuervo, A.M., Seglen, P.O.: Methods for monitoring autophagy from yeast to human. Autophagy 3(3), 181–206 (2007)

    Article  Google Scholar 

  30. Klionsky, D.J., Abdalla, F.C., Abeliovich, H., Abraham, R.T., Acevedo-Arozena, A., Adeli, K., Agholme, L., Agnello, M., Agostinis, P., Aguirre-Ghiso, J.A., et al.: Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 8(4), 445–544 (2012)

    Article  Google Scholar 

  31. Kroemer, G., Levine, B.: Autophagic cell death: the story of a misnomer. Nat. Rev. Mol. Cell Biol. 9(12), 1004–1010 (2008)

    Article  Google Scholar 

  32. Liang, C.: Negative regulation of autophagy. Cell Death Differ. 17(12), 1807–1815 (2010)

    Article  Google Scholar 

  33. Liu, L., Xie, R., Nguyen, S., Ye, M., McKeehan, W.L.: Robust autophagy/mitophagy persists during mitosis. Cell Cycle 8(10), 1616–1620 (2009)

    Article  Google Scholar 

  34. Loewer, A., Lahav, G.: We are all individuals: causes and consequences of non-genetic heterogeneity in mammalian cells. Curr. Opin. Genet. Dev. 21(6), 753–758 (2011)

    Article  Google Scholar 

  35. Loos, B., Engelbrecht, A.-M.: Cell death: a dynamic response concept. Autophagy 5(5), 590–603 (2009)

    Article  Google Scholar 

  36. Lum, J.J., Bauer, D.E., Kong, M., Harris, M.H., Li, C., Lindsten, T., Thompson, C.B.: Growth factor regulation of autophagy and cell survival in the absence of apoptosis. Cell 120(2), 237–248 (2005)

    Article  Google Scholar 

  37. Maiuri, M.C., Zalckvar, E., Kimchi, A., Kroemer, G.: Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nat. Rev. Mol. Cell Biol. 8(9), 741–752 (2007)

    Article  Google Scholar 

  38. Mizushima, N., Levine, B., Cuervo, A.M., Klionsky, D.J.: Autophagy fights disease through cellular self-digestion. Nature 451(7182), 1069–1075 (2008)

    Article  Google Scholar 

  39. Ogata, M., Hino, S.-I., Saito, A., Morikawa, K., Kondo, S., Kanemoto, S., Murakami, T., Taniguchi, M., Tanii, I., Yoshinaga, K., et al.: Autophagy is activated for cell survival after endoplasmic reticulum stress. Mol. Cell. Biol. 26(24), 9220–9231 (2006)

    Article  Google Scholar 

  40. Pankiv, S., Clausen, T.H., Lamark, T., Brech, A., Bruun, J.-A., Outzen, H., Øvervatn, A., Bjørkøy, G., Johansen, T.: P62/SQSTM1 binds directly to ATG8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J. Biol. Chem. 282(33), 24131–24145 (2007)

    Article  Google Scholar 

  41. Proikas-Cezanne, T., Codogno, P.: A new fluorescence-based assay for autophagy. Chem. Biol. 18(8), 940–941 (2011)

    Article  Google Scholar 

  42. Purvis, J.E., Lahav, G.: Encoding and decoding cellular information through signaling dynamics. Cell 152(5), 945–956 (2013)

    Article  Google Scholar 

  43. Rubinsztein, D.C.: CDKS regulate autophagy via VPS34. Mol. Cell 38(4), 483–484 (2010)

    Article  Google Scholar 

  44. Saraste, A.: Morphologic criteria and detection of apoptosis. Herz 24(3), 189–195 (1999)

    Article  Google Scholar 

  45. Sgonc, R., Wick, G.: Methods for the detection of apoptosis. Int. Arch. Allergy Immunol. 105(4), 327–332 (2009)

    Article  Google Scholar 

  46. Shvets, E., Fass, E., Elazar, Z.: Utilizing flow cytometry to monitor autophagy in living mammalian cells. Autophagy 4(5), 621–628 (2008)

    Article  Google Scholar 

  47. Spencer, S.L., Sorger, P.K.: Measuring and modeling apoptosis in single cells. Cell 144(6), 926–939 (2011)

    Article  Google Scholar 

  48. Tanida, I., Minematsu-Ikeguchi, N., Ueno, T., Kominami, E.: Lysosomal turnover, but not a cellular level, of endogenous LC3 is a marker for autophagy. Autophagy 1(2), 84–91 (2005)

    Article  Google Scholar 

  49. Tasdemir, E., Maiuri, M.C., Tajeddine, N., Vitale, I., Criollo, A., Vicencio, J.M., Hickman, J.A., Geneste, O., Kroemer, G.: Cell cycle-dependent induction of autophagy, mitophagy and reticulophagy. Cell Cycle 6(18), 2263–2267 (2007)

    Article  Google Scholar 

  50. Tyson, J.J., Baumann, W.T., Chen, C., Verdugo, A., Tavassoly, I., Wang, Y., Weiner, L.M., Clarke, R.: Dynamic modelling of oestrogen signalling and cell fate in breast cancer cells. Nat. Rev. Cancer 11(7), 523–532 (2011)

    Article  Google Scholar 

  51. Vermes, I., Haanen, C., Steffens-Nakken, H., Reutellingsperger, C.: A novel assay for apoptosis flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin v. J. Immunol. Methods 184(1), 39–51 (1995)

    Article  Google Scholar 

  52. Wang, R.C., Levine, B.: Autophagy in cellular growth control. FEBS Lett. 584(7), 1417–1426 (2010)

    Article  Google Scholar 

  53. Wells, W.A.: Autophagy unveiled. J. Cell Biol. 168(5), 676–677 (2005)

    Google Scholar 

  54. Wrighton, K.H.: Autophagy: kinase ups and downs. Nat. Rev. Mol. Cell Biol. 11(7), 464 (2010)

    Google Scholar 

  55. Xu, Y., Yuan, J., Lipinski, M.M.: Live imaging and single-cell analysis reveal differential dynamics of autophagy and apoptosis. Autophagy 9(9), 0–1 (2013)

    Google Scholar 

  56. Yao, K.C., Komata, T., Kondo, Y., Kanzawa, T., Kondo, S., Germano, I.M.: Molecular response of human glioblastoma multiforme cells to ionizing radiation: cell cycle arrest, modulation of cyclin-dependent kinase inhibitors, and autophagy. J. Neurosurg. 98(2), 378–384 (2003)

    Article  Google Scholar 

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  1. Program of Genetics, Bioinformatics and Computational Biology (GBCB), Virginia Polytechnic Institute and State University, Blacksburg, VA, USA

    Iman Tavassoly

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  1. Iman Tavassoly
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Tavassoly, I. (2015). An Experimental Framework to Study the Dynamics of Autophagic Response. In: Dynamics of Cell Fate Decision Mediated by the Interplay of Autophagy and Apoptosis in Cancer Cells. Springer Theses. Springer, Cham. https://doi.org/10.1007/978-3-319-14962-2_3

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