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Midbody accumulation through evasion of autophagy contributes to cellular reprogramming and tumorigenicity

Nature Cell Biology volume 13, pages 1214–1223 (2011)Cite this article

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A Corrigendum to this article was published on 01 December 2011

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Abstract

The midbody is a singular organelle formed between daughter cells during cytokinesis and required for their final separation. Midbodies persist in cells long after division as midbody derivatives (MBds), but their fate is unclear. Here we show that MBds are inherited asymmetrically by the daughter cell with the older centrosome. They selectively accumulate in stem cells, induced pluripotent stem cells and potential cancer ‘stem cells’ in vivo and in vitro. MBd loss accompanies stem-cell differentiation, and involves autophagic degradation mediated by binding of the autophagic receptor NBR1 to the midbody protein CEP55. Differentiating cells and normal dividing cells do not accumulate MBds and possess high autophagic activity. Stem cells and cancer cells accumulate MBds by evading autophagosome encapsulation and exhibit low autophagic activity. MBd enrichment enhances reprogramming to induced pluripotent stem cells and increases the in vitro tumorigenicity of cancer cells. These results indicate unexpected roles for MBds in stem cells and cancer ‘stem cells’.

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Figure 1: MBds accumulate within cells.
Figure 2: MBds are preferentially inherited by the cell with the older centrosome.
Figure 3: MBds accumulate in stem cells in vivo and in vitro.
Figure 4: MBd accumulation is high in stem cells and subpopulations of cancer cells and does not correlate with cell doubling time.
Figure 5: MBds in stem and cancer cells evade membrane encapsulation and lysosomal degradation.
Figure 6: Autophagy controls intracellular MBd levels.
Figure 7: NBR1 is a receptor for targeting MBds to the autophagy pathway.
Figure 8: MBd enrichment increases reprogramming efficiency and enhances in vitro tumorigenicity.

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  • 15 November 2011

    In the version of this article initially published online, the first sentence in the Acknowledgements section was incorrect. This error has been corrected in the HTML and PDF versions of the article.

References

  1. Eggert, U. S., Mitchison, T. J. & Field, C. M. Animal cytokinesis: from parts list to mechanisms. Annu. Rev. Biochem. 75, 543–66 (2006).

    Article  CAS  Google Scholar 

  2. Neumüller, R. A. & Knoblich, J. A. Dividing cellular asymmetry: asymmetric cell division and its implications for stem cells and cancer. Genes Dev. 23, 2675–2699 (2009).

    Article  CAS  Google Scholar 

  3. Doxsey, S., McCollum, D. & Theurkauf, W. Centrosomes in cellular regulation. Annu. Rev. Cell Dev. Biol. 21, 411–434 (2005).

    Article  CAS  Google Scholar 

  4. Yamashita, Y. M., Mahowald, A. P., Perlin, J. R. & Fuller, M. T. Asymmetric inheritance of mother versus daughter centrosome in stem cell division. Science 315, 518–521 (2007).

    Article  CAS  Google Scholar 

  5. Wang, X. et al. Asymmetric centrosome inheritance maintains neural progenitors in the neocortex. Nature 461, 947–955 (2009).

    Article  CAS  Google Scholar 

  6. Yamashita, Y. M., Jones, D. L. & Fuller, M. T. Orientation of asymmetric stem cell division by the APC tumor suppressor and centrosome. Science 301, 1547–1550 (2003).

    Article  CAS  Google Scholar 

  7. Barr, F. A. & Gruneberg, U. Cytokinesis: placing and making the final cut. Cell 131, 847–860 (2007).

    Article  CAS  Google Scholar 

  8. Mullins, J. M. & Biesele, J. J. Terminal phase of cytokinesis in D-98s cells. J. Cell Biol. 73, 672–684 (1977).

    Article  CAS  Google Scholar 

  9. Gromley, et al. Centriolin anchoring of exocyst and SNARE complexes at the midbody is required for secretory-vesicle-mediated abscission. Cell 123, 75–87 (2005).

    Article  CAS  Google Scholar 

  10. Steigemann, P. et al. Aurora B-mediated abscission checkpoint protects against tetraploidization. Cell 136, 473–484 (2009).

    Article  CAS  Google Scholar 

  11. Goss, J. W. & Toomre, D. K. Both daughter cells traffic and exocytose membrane at the cleavage furrow during mammalian cytokinesis. J. Cell Biol. 181, 1047–1054 (2008).

    Article  Google Scholar 

  12. Pohl, C. & Jentsch, S. Midbody ring disposal by autophagy is a post-abscission event of cytokinesis. Nat. Cell Biol. 11, 65–70 (2009).

    Article  Google Scholar 

  13. Marzesco, A. M. et al. Release of extracellular membrane particles carrying the stem cell marker prominin-1 (CD133) from neural progenitors and other epithelial cells. J. Cell Sci. 118, 2849–2858 (2005).

    Article  CAS  Google Scholar 

  14. Dubreuil, V. et al. Midbody and primary cilium of neural progenitors release extracellular membrane particles enriched in the stem cell marker prominin-1. J. Cell. Biol. 176, 483–495 (2007).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  16. Mizushima, N. & Klionsky, D. Protein turnover via autophagy: implications for metabolism. Annu. Rev. Nutr. 27, 19–40 (2007).

    Article  CAS  Google Scholar 

  17. Yorimitsu, T. & Klionsky, D. J. Eating the endoplasmic reticulum: quality control by autophagy. Trends Cell Biol. 17, 279–285 (2007).

    Article  CAS  Google Scholar 

  18. Levine, B. & Kroemer, G. Autophagy in the pathogenesis of disease. Cell 132, 27–42 (2008).

    Article  CAS  Google Scholar 

  19. Kuma, A. et al. The role of autophagy during the early neonatal starvation period. Nature 432, 1032–1036 (2004).

    Article  CAS  Google Scholar 

  20. Fimia, G. M. et al. Ambra1 regulates autophagy and development of the nervous system. Nature 447, 1121–1125 (2007).

    Article  CAS  Google Scholar 

  21. Tsukamoto, S. et al. Autophagy is essential for preimplantation development of mouse embryos. Science 321, 117–120 (2008).

    Article  CAS  Google Scholar 

  22. Cecconi, F. & Levine, B. The role of autophagy in mammalian development: cell makeover rather than cell death. Dev. Cell 15, 344–357 (2008).

    Article  CAS  Google Scholar 

  23. Hara, T. et al. Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice. Nature 441, 885–889 (2006).

    Article  CAS  Google Scholar 

  24. Komatsu, M. et al. Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice. J. Cell Biol. 169, 425–434 (2005).

    Article  CAS  Google Scholar 

  25. Rujano, M. A. et al. Polarised asymmetric inheritance of accumulated protein damage in higher eukaryotes. PLoS Biol. 4, 2325–2335 (2006).

    Article  CAS  Google Scholar 

  26. Johnston, J. A., Illing, M. E. & Kopito, R. R. Cytoplasmic dynein/dynactin mediates the assembly of aggresomes. Cell. Motil. Cytoskeleton 53, 26–38 (2002).

    Article  CAS  Google Scholar 

  27. Anderson, C. T. & Stearns, T. Centriole age underlies asynchronous primary cilium growth in mammalian cells. Curr. Biol. 19, 1498–1502 (2009).

    Article  CAS  Google Scholar 

  28. Piel, M., Meyer, P., Khodjakov, A., Rieder, C. L. & Bornens, M. The respective contributions of the mother and daughter centrioles to centrosome activity and behavior in vertebrate cells. J. Cell Biol. 149, 317–330 (2000).

    Article  CAS  Google Scholar 

  29. Oatley, J. M. & Brinster, R. L. Regulation of spermatogonial stem cell self-renewal in mammals. Annu. Rev. Cell Dev. Biol. 24, 263–286 (2008).

    Article  CAS  Google Scholar 

  30. Barroca, V. et al. Mouse differentiating spermatogonia can generate germinal stem cells in vivo. Nat. Cell Biol. 11, 190–196 (2009).

    Article  CAS  Google Scholar 

  31. Bilgüvar, K. et al. Whole-exome sequencing identifies recessive WDR62 mutations in severe brain malformations. Nature 467, 207–210 (2010).

    Article  CAS  Google Scholar 

  32. Morris, R. J. et al. Capturing and profiling adult hair follicle stem cells. Nat. Biotechnol. 22, 411–417 (2004).

    Article  CAS  Google Scholar 

  33. Conboy, M. J., Cerletti, M., Wagers, A. J. & Conboy, I. M. Immuno-analysis and FACS sorting of adult muscle fiber-associated stem/precursor cells. Methods Mol. Biol. 621, 165–173 (2010).

    Article  CAS  Google Scholar 

  34. Park, I. H. et al. Reprogramming of human somatic cells to pluripotency with defined factors. Nature 451, 141–146 (2008).

    Article  CAS  Google Scholar 

  35. Chan, E. M. et al. Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells. Nat. Biotechnol. 27, 1033–1037 (2009).

    Article  CAS  Google Scholar 

  36. Zwaka, T. P. & Thomson, J. A. Homologous recombination in human embryonic stem cells. Nat. Biotechnol. 21, 319–321 (2003).

    Article  CAS  Google Scholar 

  37. Visvader, J. E. & Lindeman, G. J. Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat. Rev. Cancer 8, 755–768 (2008).

    Article  CAS  Google Scholar 

  38. O’Brien, C. A., Pollett, A., Gallinger, S. & Dick, J. E. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445, 106–110 (2007).

    Article  CAS  Google Scholar 

  39. Pece, S. et al. Biological and molecular heterogeneity of breast cancers correlates with their cancer stem cell content. Cell 140, 62–73 (2010).

    Article  CAS  Google Scholar 

  40. Pardal, R., Clarke, M. F. & Morrison, S. J. Applying the principles of stem-cell biology to cancer. Nat. Rev. Cancer 3, 895–902 (2003).

    Article  CAS  Google Scholar 

  41. Salic, A. & Mitchison, T. J. A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc. Natl Acad. Sci. USA 105, 2415–2420 (2008).

    Article  Google Scholar 

  42. Lorenz, H., Hailey, D. W. & Lippincott-Schwartz, J. Fluorescence protease protection of GFP chimeras to reveal protein topology and subcellular localization. Nat. Methods 3, 205–210 (2006).

    Article  CAS  Google Scholar 

  43. Eskelinen, E. L., Tanaka, Y. & Saftig, P. At the acidic edge: emerging functions for lysosomal membrane proteins. Trends Cell Biol. 13, 137–145 (2003).

    Article  CAS  Google Scholar 

  44. Klionsky, D. J. et al. Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy 4, 151–175 (2008).

    Article  CAS  Google Scholar 

  45. Liang, X. H. et al. Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature 402, 672–676 (1999).

    Article  CAS  Google Scholar 

  46. Sato, K. et al. Autophagy is activated in colorectal cancer cells and contributes to the tolerance to nutrient deprivation. Cancer Res. 67, 9677–9684 (2007).

    Article  CAS  Google Scholar 

  47. Sarkar, S. et al. A rational mechanism for combination treatment of Huntington’s disease using lithium and rapamycin. Hum. Mol. Genet. 17, 170–178 (2008).

    Article  CAS  Google Scholar 

  48. Sarkar, S. et al. Lithium induces autophagy by inhibiting inositol monophosphatase. J. Cell Biol. 170, 1101–1111 (2005).

    Article  CAS  Google Scholar 

  49. Mizushima, N. & Yoshimori, T. How to interpret LC3 immunoblotting. Autophagy 3, 542–545 (2007).

    Article  CAS  Google Scholar 

  50. Bjorkoy, G. et al. p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J. Cell Biol. 171, 603–614 (2005).

    Article  CAS  Google Scholar 

  51. Komatsu, M. et al. Homeostatic levels of p62 control cytoplasmic inclusion body formation in autophagy-deficient mice. Cell 131, 1149–1163 (2007).

    Article  CAS  Google Scholar 

  52. Pankiv, S. et al. p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of of ubiquitinated protein aggregates by autophagy. J. Biol. Chem. 282, 24131–24145 (2007).

    Article  CAS  Google Scholar 

  53. Kirkin, V. et al. A role for NBR1 in autophagosomal degradation of ubiquitinated substrates. Mol. Cell 27, 505–516 (2009).

    Article  Google Scholar 

  54. Waters, S. et al. Interactions with LC3 and polyubiquitin chains link nbr1 to autophagic protein turnover. FEBS Lett. 583, 1846–1852 (2009).

    Article  CAS  Google Scholar 

  55. Yu, J. et al. Induced pluripotent stem cell lines derived from human somatic cells. Science 318, 1917–1920 (2007).

    Article  CAS  Google Scholar 

  56. Engelmann, K., Shen, H. & Finn, O. J. MCF7 side population cells with characteristics of cancer stem/progenitor cells express the tumor antigen MUC1. Cancer Res. 68, 2419–2426 (2008).

    Article  CAS  Google Scholar 

  57. Zhang, Y. et al. SEPA-1 mediates the specific recognition and degradation of P granule components by autophagy in C. elegans. Cell 136, 308–321 (2009).

    Article  CAS  Google Scholar 

  58. Strome, S. Specification of the germ line. WormBook 28, 1–10 (2005).

    Google Scholar 

  59. Fuchs, E. The tortoise and the hair: slow-cycling cells in the stem cell race. Cell 137, 811–819 (2009).

    Article  CAS  Google Scholar 

  60. Lamark, T. et al. Interaction codes within the family of mammalian Phox and Bem1p domain-containing proteins. J. Biol. Chem. 278, 34568–34581 (2003).

    Article  CAS  Google Scholar 

  61. Majeski, A. E. & Dice, J. F. Mechanisms of chaperone-mediated autophagy. Int. J. Biochem. Cell Biol. 36, 2435–2444 (2004).

    Article  CAS  Google Scholar 

  62. Kaushik, et al. Constitutive activation of chaperone-mediated autophagy in cells with impaired macroautophagy. Mol. Biol. Cell 19, 2179–2192 (2008).

    Article  CAS  Google Scholar 

  63. Nedelsky, N. et al. Autophagy and the ubiquitin–proteasome system: collaborators in neuroprotection. Biochim. Biophys. Acta. 1782, 691–699 (2008).

    Article  CAS  Google Scholar 

  64. Xu, P. & Davis, R. J. c-Jun NH2-terminal kinase is required for lineage-specific differentiation but not stem cell self-renewal. Mol. Cell Biol. 30, 1329–1340 (2010).

    Article  CAS  Google Scholar 

  65. Greenbaum, M. P., Ma, L. & Matzuk, M. M. Conversion of midbodies into germ cell intercellular bridges. Dev. Biol. 305, 389–396 (2007).

    Article  CAS  Google Scholar 

  66. Mitchison, T., Evans, L., Schulze, E. & Kirschner, M. Sites of microtubule assembly and disassembly in the mitotic spindle. Cell 45, 515–527 (1986).

    Article  CAS  Google Scholar 

  67. Yu, L. et al. Regulation of an ATG7-beclin 1 program of autophagic cell death by caspase-8. Science 304, 1500–1502 (2004).

    Article  CAS  Google Scholar 

  68. Loewer, S. et al. Large intergenic non-coding RNA-RoR modulates reprogramming of human induced pluripotent stem cells. Nat. Genet. 42, 1113–1117 (2010).

    Article  CAS  Google Scholar 

  69. Yu, J. et al. Human induced pluripotent stem cells free of vector and transgene sequences. Science 324, 797–801 (2009).

    Article  CAS  Google Scholar 

  70. Sachdev, S., Bu, Y. & Gelman, I. H. Paxillin-Y118 phosphorylation contributes to the control of Src-induced anchorage-independent growth by FAK and adhesion. BMC Cancer 12, 9–12 (2009).

    Google Scholar 

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Acknowledgements

We thank E. Baehrecke for critical reading of the manuscript, T. Schlaeger and colleagues for assistance with H1-OGN and associated cell lines, the University of Massachusetts Medical School (UMMS) Flow Facility for assistance with MBd enrichment, P. Furcinitti of UMMS Digital Light Microscopy Core Facility for assistance with imaging, the UMMS DERC Morphology Core for assistance with immunohistochemitry, D. Guertin and C. Sparks for assistance with SMP preparation, S. Lyle and C. Powers for sample preparation and H-L. Liu for assistance with clone construction. We thank N. Mizushima for GFP–LC3-expressing Atg5−/− and matchedwild-type MEFs, M. Komatsu and T. Ishii for p 62−/− and matched wild-type MEFs, S. Jones for ex vivo C57BL/6 MEFs, B. Lewis for mouse hepatocellular cancer lines, S. Pino for in vitro activated T cells, W. Jiang for MKLP1–GFP plasmid, K. Khanna for CEP55–eGFP plasmid, J. Lippincott-Schwartz and G. Gaietta for plasmids for FPP assay, B. Levine (UT Southwestern) for Flag-tagged BECN1-expressing plasmid, A. Khodjakov for CETN1–GFP-expressing plasmids, K. Lee for hCenexin1 antibody and the Progeria Society for cell lines. The α6F antibody to Na/K-ATPase developed by D. M. Fambrough and the H4B4 antibody to LAMP2 developed by J.T. August and J. E. K. Hildreth were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the National Institute of Child Health and Human Development and maintained by The University of Iowa, Department of Biology. This work was supported by funding from the National Institutes of Health (GM051994 to S.D. and F32 GM084660-02 to D.B.), the W.M. Keck Foundation to S.D., the Ellison Foundation (AG-SS-1918-07) to S.D., the Department of Defense (W81XWH-08-1-0457 to S.D. and W81XWH-06-1-0140 to C-T.C.) and the Diabetes and Endocrine Resource Center (5P30DK3252025). Core resources supported by the Diabetes Endocrinology Research Center grant DK32520 were also used.

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  1. Tse-Chun Kuo and Chun-Ting Chen: These authors contributed equally to this work

Authors and Affiliations

  1. Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA

    Tse-Chun Kuo, Chun-Ting Chen, Desiree Baron, Cara M. Weismann, Ping Xu & Stephen Doxsey

  2. Stem Cell Program, Children’s Hospital Boston, Boston, Massachusetts 02115, USA

    Tamer T. Onder, Sabine Loewer & George Q. Daley

  3. Division of Pediatric Hematology and Oncology, Stem Cell Transplantation Program, Manton Center for Orphan Disease Research, Children’s Hospital Boston and Dana Farber Cancer Institute, Boston, Massachusetts 02115, USA

    Tamer T. Onder, Sabine Loewer & George Q. Daley

  4. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA

    Tamer T. Onder, Sabine Loewer & George Q. Daley

  5. Harvard Stem Cell Institute, Cambridge, Massachusetts 02138, USA

    Tamer T. Onder, Sabine Loewer & George Q. Daley

  6. Departments of Neurology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA

    Sandra Almeida, Cara M. Weismann & Fen-Biao Gao

  7. Department of Pathology, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA

    Jean-Marie Houghton

  8. Howard Hughes Medical Institute, Chevy Chase, Maryland 20815, USA

    George Q. Daley

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Contributions

C-T.C. and S.D. conceived the project and wrote the manuscript with the help of T-C.K. and D.B. The experiments on the inheritance and localization of MBds as well as some for MBd degradation were conducted by C-T.C. The experiments on MBd accumulation were conducted by C-T.C. with the help of T-C.K. and C.M.W. Investigation of the mechanisms for MBd degradation was conceived by T-C.K. and S.D., and much of the work executed by T-C.K. Autophagic flux assay, soft-agar assay of FACS-isolated cells and MBd localization in neural progenitors were conducted by D.B., who contributed substantially to the work and intellectual input on multiple aspects of this project. The reprogramming assay was conducted and analysed by T-C.K., T.T.O and S.L. The preparation of hESCs for live imaging was conducted by S.A. Tissue preparation was assisted by P.X. and J.M.H.

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Correspondence to Stephen Doxsey.

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Kuo, TC., Chen, CT., Baron, D. et al. Midbody accumulation through evasion of autophagy contributes to cellular reprogramming and tumorigenicity. Nat Cell Biol 13, 1214–1223 (2011). https://doi.org/10.1038/ncb2332

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