Abstract
Autophagy is a catabolic process in which cell components are degraded to maintain cellular homeostasis by nutrient limitations. Defects of autophagy are involved in numerous diseases, including cancer. Here, we demonstrate a new role of phospholipase D (PLD) as a regulator of autophagy. PLD inhibition enhances autophagic flux via ATG1 (ULK1), ATG5 and ATG7, which are essential autophagy gene products critical for autophagosome formation. Moreover, PLD suppresses autophagy by differentially modulating phosphorylation of ULK1 mediated by mTOR and adenosine monophosphate-activated protein kinase (AMPK), and by suppressing the interaction of Beclin 1 with vacuolar-sorting protein 34 (Vps34), indicating that PLD coordinates major players of the autophagic pathway, AMPK-mTOR-ULK1 and Vps34/Beclin 1. Ultimately, PLD inhibition significantly sensitized in vitro and in vivo cancer regression via genetic and pharmacological inhibition of autophagy, providing rationale for a new therapeutic approach to enhancing the anticancer efficacy of PLD inhibition. Collectively, we show a novel role for PLD in the molecular machinery regulating autophagy.
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
- PLD:
-
phospholipase D
- PA:
-
phosphatidic acid
- AMPK:
-
adenosine monophosphate-activated protein kinase
- LC3:
-
microtubule-associated protein light chain 3
- Vps34:
-
vacuolar-sorting protein 34
- CQ:
-
chloroquine
- Baf A1:
-
bafilomycin A1
- ABC:
-
avidin-biotinylated enzyme complex
- DAB:
-
3,3-diaminobenzidine
- mRFP:
-
monomeric red fluorescent protein
- MEF:
-
mouse embryonic fibroblast
- PI:
-
propidium iodide
- TEM:
-
transmission electron microscopy
References
Mathew R, Karantza-Wadsworth V, White E . Role of autophagy in cancer. Nat Rev Cancer 2007; 7: 961–967.
Tanida I, Ueno T, Kominami E . LC3 conjugation system in mammalian autophagy. Int J Biochem Cell Biol 2004; 36: 2503–2518.
Mizushima N, Yoshimori T, Levine B . Methods in mammalian autophagy research. Cell 2010; 140: 313–326.
Kuma A, Hatano M, Matsui M, Yamamoto A, Nakaya H, Yoshimori T et al. The role of autophagy during the early neonatal starvation period. Nature 2004; 432: 1032–1036.
Levine B, Kroemer G . Autophagy in the pathogenesis of disease. Cell 2008; 132: 27–42.
Chen N, Debnath J . Autophagy and tumorigenesis. FEBS Lett 2010; 584: 1427–1435.
Degenhardt K, Mathew R, Beaudoin B, Bray K, Anderson D, Chen G et al. Autophagy promotes tumor cell survival and restricts necrosis, inflammation, and tumorigenesis. Cancer Cell 2006; 10: 51–64.
Chen N, Karantza V . Autophagy as a therapeutic target in cancer. Cancer Biol Ther 2011; 11: 157–168.
Kuma A, Mizushima N . Physiological role of autophagy as an intracellular recycling system: with an emphasis on nutrient metabolism. Semin Cell Dev Biol 2010; 21: 683–690.
Efeyan A, Sabatini DM . mTOR and cancer: many loops in one pathway. Curr Opin Cell Biol 2010; 22: 169–176.
Kim J, Kundu M, Viollet B, Guan KL . AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol 2011; 13: 132–141.
Egan DF, Shackelford DB, Mihaylova MM, Gelino S, Kohnz RA, Mair W et al. Phosphorylation of ULK1 (hATG1) by AMP-activated protein kinase connects energy sensing to mitophagy. Science 2011; 331: 456–461.
Funderburk SF, Wang QJ, Yue Z . The Beclin 1-VPS34 complex—at the crossroads of autophagy and beyond. Trends Cell Biol 2010; 20: 355–362.
Pattingre S, Tassa A, Qu X, Garuti R, Liang XH, Mizushima N et al. Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell 2005; 122: 927–939.
Wei Y, Pattingre S, Sinha S, Bassik M, Levine B . JNK1-mediated phosphorylation of Bcl-2 regulates starvation-induced autophagy. Mol Cell 2008; 30: 678–688.
Foster DA . Regulation of mTOR by phosphatidic acid? Cancer Res 2007; 67: 1–4.
Sun Y, Chen J . mTOR signaling: PLD takes center stage. Cell Cycle 2008; 7: 3118–3123.
Kang DW, Choi KY, Min do S . Phospholipase D meets Wnt signaling: a new target for cancer therapy. Cancer Res 2011; 71: 293–297.
Huang P, Frohman MA . The potential for phospholipase D as a new therapeutic target. Expert Opin Ther Targets 2007; 11: 707–716.
Mathew R, Karp CM, Beaudoin B, Vuong N, Chen G, Chen HY et al. Autophagy suppresses tumorigenesis through elimination of p62. Cell 2009; 137: 1062–1075.
Kimura S, Noda T, Yoshimori T . Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy 2007; 3: 452–460.
Farkas T, Hoyer-Hansen M, Jaattela M . Identification of novel autophagy regulators by a luciferase-based assay for the kinetics of autophagic flux. Autophagy 2009; 5: 1018–1025.
Frankel LB, Wen J, Lees M, Hoyer-Hansen M, Farkas T, Krogh A et al. microRNA-101 is a potent inhibitor of autophagy. EMBO J 2011; 30: 4628–4641.
Hosokawa N, Hara Y, Mizushima N . Generation of cell lines with tetracycline-regulated autophagy and a role for autophagy in controlling cell size. FEBS Lett 2007; 581: 2623–2629.
Komatsu M, Waguri S, Ueno T, Iwata J, Murata S, Tanida I et al. Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice. J Cell Biol 2005; 169: 425–434.
Faivre S, Kroemer G, Raymond E . Current development of mTOR inhibitors as anticancer agents. Nat Rev Drug Discov 2006; 5: 671–688.
Meley D, Bauvy C, Houben-Weerts JH, Dubbelhuis PF, Helmond MT, Codogno P et al. AMP-activated protein kinase and the regulation of autophagic proteolysis. J Biol Chem 2006; 281: 34870–34879.
Kang R, Zeh HJ, Lotze MT, Tang D . The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ 2011; 18: 571–580.
Zeng X, Overmeyer JH, Maltese WA . Functional specificity of the mammalian Beclin-Vps34 PI 3-kinase complex in macroautophagy versus endocytosis and lysosomal enzyme trafficking. J Cell Sci 2006; 119: 259–270.
Furuya N, Yu J, Byfield M, Pattingre S, Levine B . The evolutionarily conserved domain of Beclin 1 is required for Vps34 binding, autophagy and tumor suppressor function. Autophagy 2005; 1: 46–52.
White E, DiPaola RS . The double-edged sword of autophagy modulation in cancer. Clin Cancer Res 2009; 15: 5308–5316.
Carew JS, Nawrocki ST, Kahue CN, Zhang H, Yang C, Chung L et al. Targeting autophagy augments the anticancer activity of the histone deacetylase inhibitor SAHA to overcome Bcr-Abl-mediated drug resistance. Blood 2007; 110: 313–322.
Yang ZJ, Chee CE, Huang S, Sinicrope FA . The role of autophagy in cancer: therapeutic implications. Mol Cancer Ther 2011; 10: 1533–1541.
Salazar M, Carracedo A, Salanueva IJ, Hernandez-Tiedra S, Lorente M, Egia A et al. Cannabinoid action induces autophagy-mediated cell death through stimulation of ER stress in human glioma cells. J Clin Invest 2009; 119: 1359–1372.
Amaravadi RK, Yu D, Lum JJ, Bui T, Christophorou MA, Evan GI et al. Autophagy inhibition enhances therapy-induced apoptosis in a Myc-induced model of lymphoma. J Clin Invest 2007; 117: 326–336.
Yang ZJ, Chee CE, Huang S, Sinicrope F . Autophagy modulation for cancer therapy. Cancer Biol Ther 2011; 11: 169–176.
Kim J, Lee YH, Kwon TK, Chang JS, Chung KC, Min DS . Phospholipase D prevents etoposide-induced apoptosis by inhibiting the expression of early growth response-1 and phosphatase and tensin homologue deleted on chromosome 10. Cancer Res 2006; 66: 784–793.
Rubinsztein DC, Cuervo AM, Ravikumar B, Sarkar S, Korolchuk V, Kaushik S et al. In search of an ‘autophagomometer’. Autophagy 2009; 5: 585–589.
Lavieu G, Scarlatti F, Sala G, Carpentier S, Levade T, Ghidoni R et al. Regulation of autophagy by sphingosine kinase 1 and its role in cell survival during nutrient starvation. J Biol Chem 2006; 281: 8518–8527.
Dall'Armi C, Hurtado-Lorenzo A, Tian H, Morel E, Nezu A, Chan R et al. The phospholipase D1 pathway modulates macroautophagy. Nat Commun 2010; 1: 142.
Feng Z, Zhang H, Levine AJ, Jin S . The coordinate regulation of the p53 and mTOR pathways in cells. Proc Natl Acad Sci USA 2005; 102: 8204–8209.
Crighton D, Wilkinson S, O'Prey J, Syed N, Smith P, Harrison PR et al. DRAM, a p53-induced modulator of autophagy, is critical for apoptosis. Cell 2006; 126: 121–134.
Tasdemir E, Maiuri MC, Galluzzi L, Vitale I, Djavaheri-Mergny M, D'Amelio M et al. Regulation of autophagy by cytoplasmic p53. Nat Cell Biol 2008; 10: 676–687.
Backer JM . The regulation and function of class III PI3Ks: novel roles for Vps34. Biochem J 2008; 410: 1–17.
Pattingre S, Espert L, Biard-Piechaczyk M, Codogno P . Regulation of macroautophagy by mTOR and Beclin 1 complexes. Biochimie 2008; 90: 313–323.
Yoon MS, Du G, Backer JM, Frohman MA, Chen J . Class III PI-3-kinase activates phospholipase D in an amino acid-sensing mTORC1 pathway. J Cell Biol 2011; 195: 435–447.
Xie Z, Klionsky DJ . Autophagosome formation: core machinery and adaptations. Nat Cell Biol 2007; 9: 1102–1109.
Nishida Y, Arakawa S, Fujitani K, Yamaguchi H, Mizuta T, Kanaseki T et al. Discovery of Atg5/Atg7-independent alternative macroautophagy. Nature 2009; 461: 654–658.
Shimizu S, Arakawa S, Nishida Y . Autophagy takes an alternative pathway. Autophagy 2010; 6: 290–291.
Degtyarev M, De Maziere A, Orr C, Lin J, Lee BB, Tien JY et al. Akt inhibition promotes autophagy and sensitizes PTEN-null tumors to lysosomotropic agents. J Cell Biol 2008; 183: 101–116.
Jang YH, Ahn BH, Namkoong S, Kim YM, Jin JK, Kim YS et al. Differential regulation of apoptosis by caspase-mediated cleavage of phospholipase D isozymes. Cell Signal 2008; 20: 2190–2207.
Jang YH, Min do S . Nuclear localization of phospholipase D1 mediates the activation of nuclear protein kinase C(alpha) and extracellular signal-regulated kinase signaling pathways. J Biol Chem 2011; 286: 4680–4689.
Acknowledgements
This study was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST; No. 2012002009) and a Translational Research Center for Protein Function Control Grant (NSF 2009-0092960). We thank Dr. T Yosihmori (Osaka University), Misushima (Tokyo Medical and Dental University), G Velasco (Complutense University), M Komatsu (Tokyo Metropolitan Institute of Medical Science), M Colombo (Universidad Nacional de Cuyo, Mendoza), M Kundu (St. Jude Children’s Hospital), B Levine (University of Texas Southwestern Medical School), G Paolo (Columbia University) and Marja Jäättelä (Apoptosis Department and Centre for Genotoxic Stress Research, Denmark) for materials.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Edited by E Baehrecke
Supplementary Information accompanies this paper on Cell Death and Differentiation website
Supplementary information
Rights and permissions
About this article
Cite this article
Jang, Y., Choi, K. & Min, D. Phospholipase D-mediated autophagic regulation is a potential target for cancer therapy. Cell Death Differ 21, 533–546 (2014). https://doi.org/10.1038/cdd.2013.174
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/cdd.2013.174
Keywords
This article is cited by
-
Autophagy and autophagy signaling in Epilepsy: possible role of autophagy activator
Molecular Medicine (2023)
-
Sam68 promotes osteogenic differentiation of aortic valvular interstitial cells by TNF-α/STAT3/autophagy axis
Journal of Cell Communication and Signaling (2023)
-
SCD1, autophagy and cancer: implications for therapy
Journal of Experimental & Clinical Cancer Research (2021)
-
Targeting phospholipase D in cancer, infection and neurodegenerative disorders
Nature Reviews Drug Discovery (2017)
-
Phospholipase D1 deficiency in mice causes nonalcoholic fatty liver disease via an autophagy defect
Scientific Reports (2016)
