Abstract
Cellular senescence is considered as an important tumor-suppressive mechanism. Here, we demonstrated that heparan sulfate (HS) prevents cellular senescence by fine-tuning of the fibroblast growth factor receptor (FGFR) signaling pathway. We found that depletion of 3′-phosphoadenosine 5′-phosphosulfate synthetase 2 (PAPSS2), a synthetic enzyme of the sulfur donor PAPS, led to premature cell senescence in various cancer cells and in a xenograft tumor mouse model. Sodium chlorate, a metabolic inhibitor of HS sulfation also induced a cellular senescence phenotype. p53 and p21 accumulation was essential for PAPSS2-mediated cellular senescence. Such senescence phenotypes were closely correlated with cell surface HS levels in both cancer cells and human diploid fibroblasts. The determination of the activation of receptors such as FGFR1, Met, and insulin growth factor 1 receptor β indicated that the augmented FGFR1/AKT signaling was specifically involved in premature senescence in a HS-dependent manner. Thus, blockade of either FGFR1 or AKT prohibited p53 and p21 accumulation and cell fate switched from cellular senescence to apoptosis. In particular, desulfation at the 2-O position in the HS chain contributed to the premature senescence via the augmented FGFR1 signaling. Taken together, we reveal, for the first time, that the proper status of HS is essential for the prevention of cellular senescence. These observations allowed us to hypothesize that the FGF/FGFR signaling system could initiate novel tumor defenses through regulating premature senescence.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- FACS:
-
Fluorescence activated cell sorter
- FGF:
-
fibroblast growth factor
- FGFR:
-
fibroblast growth factor receptor
- HDF:
-
human diploid fibroblast
- HGF:
-
hepatocyte growth factor
- HS6ST1:
-
heparan sulfate 6-O-sulfotransferase 1
- HSPG:
-
heparan sulfate proteoglycan
- IGF1Rβ:
-
insulin-like growth factor 1 receptor β
- IP:
-
immunoprecipitation
- KI:
-
kinase inactive
- PAPSS2:
-
3′-phosphoadenosine 5′-phosphosulfate synthetase 2
- PARP:
-
poly ADP ribose polymerase
- PG:
-
proteoglycan
- SA-β-Gal:
-
senescence-associated β-galactosidase
- SASP:
-
senescence associated secretory phenotype
- WB:
-
western blot
- WT:
-
wild type
References
Narita M, Lowe SW . Senescence comes of age. Nat Med 2005; 11: 920–922.
Campisi J . Aging, cellular senescence, and cancer. Annu Rev Physiol 2013; 75: 685–705.
Collado M, Serrano M . Senescence in tumours: evidence from mice and humans. Nat Rev Cancer 2010; 10: 51–57.
Campisi J, d'Adda di Fagagna F . Cellular senescence: when bad things happen to good cells. Nat Rev Mol Cell Biol 2007; 8: 729–740.
Prieur A, Peeper DS . Cellular senescence in vivo: a barrier to tumorigenesis. Curr Opin Cell Biol 2008; 20: 150–155.
Ohtani N, Mann DJ, Hara E . Cellular senescence: its role in tumor suppression and aging. Cancer Sci 2009; 100: 792–797.
Nogueira V, Park Y, Chen CC, Xu PZ, Chen ML, Tonic I et al. Akt determines replicative senescence and oxidative or oncogenic premature senescence and sensitizes cells to oxidative apoptosis. Cancer Cell 2008; 14: 458–470.
Shay JW, Roninson IB . Hallmarks of senescence in carcinogenesis and cancer therapy. Oncogene 2004; 23: 2919–2933.
Schmitt CA . Cellular senescence and cancer treatment. Biochim Biophys Acta 2007; 1775: 5–20.
Lee M, Lee JS . Exploiting tumor cell senescence in anticancer therapy. BMB Rep 2014; 47: 51–59.
Kuilman T, Peeper DS . Senescence-messaging secretome: SMS-ing cellular stress. Nat Rev Cancer 2009; 9: 81–94.
Coppe JP, Desprez PY, Krtolica A, Campisi J . The senescence-associated secretory phenotype: the dark side of tumor suppression. Annu Rev Pathol 2010; 5: 99–118.
Rodier F, Campisi J . Four faces of cellular senescence. J Cell Biol 2011; 192: 547–556.
Perez-Mancera PA, Young AR, Narita M . Inside and out: the activities of senescence in cancer. Nat Rev Cancer 2014; 14: 547–558.
Campisi J . Cellular senescence: putting the paradoxes in perspective. Curr Opin Genet Dev 2011; 21: 107–112.
Munoz-Espin D, Canamero M, Maraver A, Gomez-Lopez G, Contreras J, Murillo-Cuesta S et al. Programmed cell senescence during mammalian embryonic development. Cell 2013; 155: 1104–1118.
Storer M, Mas A, Robert-Moreno A, Pecoraro M, Ortells MC, Di Giacomo V et al. Senescence is a developmental mechanism that contributes to embryonic growth and patterning. Cell 2013; 155: 1119–1130.
Bernfield M, Gotte M, Park PW, Reizes O, Fitzgerald ML, Lincecum J et al. Functions of cell surface heparan sulfate proteoglycans. Annu Rev Biochem 1999; 68: 729–777.
Hacker U, Nybakken K, Perrimon N . Heparan sulphate proteoglycans: the sweet side of development. Nat Rev Mol Cell Biol 2005; 6: 530–541.
Bishop JR, Schuksz M, Esko JD . Heparan sulphate proteoglycans fine-tune mammalian physiology. Nature 2007; 446: 1030–1037.
Couchman JR . Transmembrane signaling proteoglycans. Annu Rev Cell Dev Biol 2010; 26: 89–114.
Turnbull J, Powell A, Guimond S . Heparan sulfate: decoding a dynamic multifunctional cell regulator. Trends Cell Biol 2001; 11: 75–82.
Li H, Deyrup A, Mensch Jr JR, Domowicz M, Konstantinidis AK, Schwartz NB . The isolation and characterization of cDNA encoding the mouse bifunctional ATP sulfurylase-adenosine 5'-phosphosulfate kinase. J Biol Chem 1995; 270: 29453–29459.
Xu ZH, Otterness DM, Freimuth RR, Carlini EJ, Wood TC, Mitchell S et al. Human 3'-phosphoadenosine 5′-phosphosulfate synthetase 1 (PAPSS1) and PAPSS2: gene cloning, characterization and chromosomal localization. Biochem Biophys Res Commun 2000; 268: 437–444.
Besset S, Vincourt JB, Amalric F, Girard JP . Nuclear localization of PAPS synthetase 1: a sulfate activation pathway in the nucleus of eukaryotic cells. FASEB J 2000; 14: 345–354.
Schroder E, Gebel L, Eremeev AA, Morgner J, Grum D, Knauer SK et al. Human PAPS synthase isoforms are dynamically regulated enzymes with access to nucleus and cytoplasm. PLoS One 2012; 7: e29559.
Esko JD, Selleck SB . Order out of chaos: assembly of ligand binding sites in heparan sulfate. Annu Rev Biochem 2002; 71: 435–471.
Kusche-Gullberg M, Kjellen L . Sulfotransferases in glycosaminoglycan biosynthesis. Curr Opin Struct Biol 2003; 13: 605–611.
Zhang L . Glycosaminoglycan (GAG) biosynthesis and GAG-binding proteins. Prog Mol Biol Transl Sci 2010; 93: 1–17.
Spivak-Kroizman T, Lemmon MA, Dikic I, Ladbury JE, Pinchasi D, Huang J et al. Heparin-induced oligomerization of FGF molecules is responsible for FGF receptor dimerization, activation, and cell proliferation. Cell 1994; 79: 1015–1024.
Yayon A, Klagsbrun M, Esko JD, Leder P, Ornitz DM . Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor. Cell 1991; 64: 841–848.
Rapraeger AC, Krufka A, Olwin BB . Requirement of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation. Science 1991; 252: 1705–1708.
Qu X, Carbe C, Tao C, Powers A, Lawrence R, van Kuppevelt TH et al. Lacrimal gland development and Fgf10-Fgfr2b signaling are controlled by 2-O- and 6-O-sulfated heparan sulfate. J Biol Chem 2011; 286: 14435–14444.
Turner N, Grose R . Fibroblast growth factor signalling: from development to cancer. Nat Rev Cancer 2010; 10: 116–129.
Safaiyan F, Kolset SO, Prydz K, Gottfridsson E, Lindahl U, Salmivirta M . Selective effects of sodium chlorate treatment on the sulfation of heparan sulfate. J Biol Chem 1999; 274: 36267–36273.
Qiu H, Jiang JL, Liu M, Huang X, Ding SJ, Wang L . Quantitative phosphoproteomics analysis reveals broad regulatory role of heparan sulfate on endothelial signaling. Mol Cell Proteomics 2013; 12: 2160–2173.
Lund J, Sondergaard MT, Conover CA, Overgaard MT . Heparin-binding mechanism of the IGF2/IGF-binding protein 2 complex. J Mol Endocrinol 2014; 52: 345–355.
Arai T, Busby Jr W, Clemmons DR . Binding of insulin-like growth factor (IGF) I or II to IGF-binding protein-2 enables it to bind to heparin and extracellular matrix. Endocrinology 1996; 137: 4571–4575.
Harmer NJ, Chirgadze D, Hyun Kim K, Pellegrini L, Blundell TL . The structural biology of growth factor receptor activation. Biophys Chem 2003; 100: 545–553.
Ashikari-Hada S, Habuchi H, Sugaya N, Kobayashi T, Kimata K . Specific inhibition of FGF-2 signaling with 2-O-sulfated octasaccharides of heparan sulfate. Glycobiology 2009; 19: 644–654.
Frese MA, Milz F, Dick M, Lamanna WC, Dierks T . Characterization of the human sulfatase Sulf1 and its high affinity heparin/heparan sulfate interaction domain. J Biol Chem 2009; 284: 28033–28044.
Nagamine S, Tamba M, Ishimine H, Araki K, Shiomi K, Okada T et al. Organ-specific sulfation patterns of heparan sulfate generated by extracellular sulfatases Sulf1 and Sulf2 in mice. J Biol Chem 2012; 287: 9579–9590.
Hayflick L, Moorhead PS . The serial cultivation of human diploid cell strains. Exp Cell Res 1961; 25: 585–621.
Byun HO, Han NK, Lee HJ, Kim KB, Ko YG, Yoon G et al. Cathepsin D and eukaryotic translation elongation factor 1 as promising markers of cellular senescence. Cancer Res 2009; 69: 4638–4647.
Lee JJ, Kim BC, Park MJ, Lee YS, Kim YN, Lee BL et al. PTEN status switches cell fate between premature senescence and apoptosis in glioma exposed to ionizing radiation. Cell Death Differ 2011; 18: 666–677.
Ben-Porath I, Weinberg RA . The signals and pathways activating cellular senescence. Int J Biochem Cell Biol 2005; 37: 961–976.
Wajapeyee N, Serra RW, Zhu X, Mahalingam M, Green MR . Oncogenic BRAF induces senescence and apoptosis through pathways mediated by the secreted protein IGFBP7. Cell 2008; 132: 363–374.
Itoh N, Ornitz DM . Fibroblast growth factors: from molecular evolution to roles in development, metabolism and disease. J Biochem 2011; 149: 121–130.
Itoh N, Ornitz DM . Evolution of the Fgf and Fgfr gene families. Trends Genet 2004; 20: 563–569.
Westwood G, Dibling BC, Cuthbert-Heavens D, Burchill SA . Basic fibroblast growth factor (bFGF)-induced cell death is mediated through a caspase-dependent and p53-independent cell death receptor pathway. Oncogene 2002; 21: 809–824.
Williamson AJ, Dibling BC, Boyne JR, Selby P, Burchill SA . Basic fibroblast growth factor-induced cell death is effected through sustained activation of p38MAPK and up-regulation of the death receptor p75NTR. J Biol Chem 2004; 279: 47912–47928.
Wang H, Rubin M, Fenig E, DeBlasio A, Mendelsohn J, Yahalom J et al. Basic fibroblast growth factor causes growth arrest in MCF-7 human breast cancer cells while inducing both mitogenic and inhibitory G1 events. Cancer Res 1997; 57: 1750–1757.
Krejci P, Bryja V, Pachernik J, Hampl A, Pogue R, Mekikian P et al. FGF2 inhibits proliferation and alters the cartilage-like phenotype of RCS cells. Exp Cell Res 2004; 297: 152–164.
Pye DA, Vives RR, Hyde P, Gallagher JT . Regulation of FGF-1 mitogenic activity by heparan sulfate oligosaccharides is dependent on specific structural features: differential requirements for the modulation of FGF-1 and FGF-2. Glycobiology 2000; 10: 1183–1192.
Leontieva OV, Blagosklonny MV . DNA damaging agents and p53 do not cause senescence in quiescent cells, while consecutive re-activation of mTOR is associated with conversion to senescence. Aging 2010; 2: 924–935.
Kolesnichenko M, Hong L, Liao R, Vogt PK, Sun P . Attenuation of TORC1 signaling delays replicative and oncogenic RAS-induced senescence. Cell Cycle 2012; 11: 2391–2401.
Xue W, Zender L, Miething C, Dickins RA, Hernando E, Krizhanovsky V et al. Senescence and tumour clearance is triggered by p53 restoration in murine liver carcinomas. Nature 2007; 445: 656–660.
Krizhanovsky V, Yon M, Dickins RA, Hearn S, Simon J, Miething C et al. Senescence of activated stellate cells limits liver fibrosis. Cell 2008; 134: 657–667.
Plotnikov AN, Schlessinger J, Hubbard SR, Mohammadi M . Structural basis for FGF receptor dimerization and activation. Cell 1999; 98: 641–650.
Carey DJ . Syndecans: multifunctional cell-surface co-receptors. Biochem J 1997; 327: 1–16.
Matsuo I, Kimura-Yoshida C . Extracellular modulation of Fibroblast Growth Factor signaling through heparan sulfate proteoglycans in mammalian development. Curr Opin Genet Dev 2013; 23: 399–407.
Richard C, Liuzzo JP, Moscatelli D . Fibroblast growth factor-2 can mediate cell attachment by linking receptors and heparan sulfate proteoglycans on neighboring cells. J Biol Chem 1995; 270: 24188–24196.
Roghani M, Mansukhani A, Dell'Era P, Bellosta P, Basilico C, Rifkin DB et al. Heparin increases the affinity of basic fibroblast growth factor for its receptor but is not required for binding. J Biol Chem 1994; 269: 3976–3984.
Ai X, Do AT, Lozynska O, Kusche-Gullberg M, Lindahl U, Emerson CP Jr . QSulf1 remodels the 6-O sulfation states of cell surface heparan sulfate proteoglycans to promote Wnt signaling. J Cell Biol 2003; 162: 341–351.
Jemth P, Kreuger J, Kusche-Gullberg M, Sturiale L, Gimenez-Gallego G, Lindahl U . Biosynthetic oligosaccharide libraries for identification of protein-binding heparan sulfate motifs. Exploring the structural diversity by screening for fibroblast growth factor (FGF)1 and FGF2 binding. J Biol Chem 2002; 277: 30567–30573.
Kiefer MC, Baird A, Nguyen T, George-Nascimento C, Mason OB, Boley LJ et al. Molecular cloning of a human basic fibroblast growth factor receptor cDNA and expression of a biologically active extracellular domain in a baculovirus system. Growth Factors 1991; 5: 115–127.
Presta M, Statuto M, Isacchi A, Caccia P, Pozzi A, Gualandris et al. Structure-function relationship of basic fibroblast growth factor: site-directed mutagenesis of a putative heparin-binding and receptor-binding region. Biochem Biophys Res Commun 1992; 185: 1098–1107.
Fujita K, Horikawa I, Mondal AM, Jenkins LM, Appella E, Vojtesek B et al. Positive feedback between p53 and TRF2 during telomere-damage signalling and cellular senescence. Nat Cell Biol 2010; 12: 1205–1212.
Biroccio A, Cherfils-Vicini J, Augereau A, Pinte S, Bauwens S, Ye J et al. TRF2 inhibits a cell-extrinsic pathway through which natural killer cells eliminate cancer cells. Nat Cell Biol 2013; 15: 818–828.
Persaud A, Alberts P, Hayes M, Guettler S, Clarke I, Sicheri F et al. Nedd4-1 binds and ubiquitylates activated FGFR1 to control its endocytosis and function. EMBO J 2011; 30: 3259–3273.
Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C et al. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci USA 1995; 92: 9363–9367.
Kim BC, Lee HC, Lee JJ, Choi CM, Kim DK, Lee JC et al. Wig1 prevents cellular senescence by regulating p21 mRNA decay through control of RISC recruitment. EMBO J 2012; 31: 4289–4303.
Acknowledgements
This work was supported by the Nuclear Research and Development Program (Grant No. 2012M2B2B1055637) and Medical Research Center (MRC) (Grant No. 2014009392) of the National Research Foundation of Korea (NRF), funded by the Korean government (MSIP).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Edited by J Bartek
Supplementary Information accompanies this paper on Cell Death and Differentiation website .
Supplementary information
Rights and permissions
About this article
Cite this article
Jung, S., Lee, H., Yu, DM. et al. Heparan sulfation is essential for the prevention of cellular senescence. Cell Death Differ 23, 417–429 (2016). https://doi.org/10.1038/cdd.2015.107
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/cdd.2015.107
This article is cited by
-
A novel long noncoding RNA Linc-ASEN represses cellular senescence through multileveled reduction of p21 expression
Cell Death & Differentiation (2020)
-
Enhanced PAPSS2/VCAN sulfation axis is essential for Snail-mediated breast cancer cell migration and metastasis
Cell Death & Differentiation (2019)
-
Integrin α6β4-Src-AKT signaling induces cellular senescence by counteracting apoptosis in irradiated tumor cells and tissues
Cell Death & Differentiation (2019)
-
mTOR kinase leads to PTEN-loss-induced cellular senescence by phosphorylating p53
Oncogene (2019)
-
Large-scale interaction effects reveal missing heritability in schizophrenia, bipolar disorder and posttraumatic stress disorder
Translational Psychiatry (2017)
