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Inhibition of Autophagy by Captopril Attenuates Prion Peptide-Mediated Neuronal Apoptosis via AMPK Activation

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Abstract

Accumulation of prion protein (PrPc) into a protease-resistant form (PrPsc) in the brains of humans and animals affects the central nervous system. PrPsc occurs only in mammals with transmissible prion diseases. Prion protein refers to either the infectious pathogen itself or the main component of the pathogen. Recent studies suggest that autophagy is one of the major functions that keep cells alive and which has a protective effect against neurodegeneration. In this study, we investigated whether the anti-hypertensive drug, captopril, could attenuate prion peptide PrP (106–126)-induced calcium alteration-mediated neurotoxicity. Treatment with captopril increased both LC3-II (microtubule-associated protein 1A/1B-light chain 3-II) and p62 protein levels, indicating autophagy flux inhibition. Electron microscopy confirmed the occurrence of autophagic flux inhibition in neuronal cells treated with captopril. Captopril attenuated PrP (106–126)-induced neuronal cell death via AMPK activation and autophagy inhibition. Compound C suppressed AMPK activation as well as the neuroprotective effects of captopril. Thus, these data showed that an anti-hypertensive drug has a protective effect against prion-mediated neuronal cell death via autophagy inhibition and AMPK activation, and also suggest that anti-hypertensive drugs may be effective therapeutic agents against neurodegenerative disorders, including prion diseases.

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References

  1. Costanzo M, Zurzolo C (2013) The cell biology of prion-like spread of protein aggregates: mechanisms and implication in neurodegeneration. Biochem J 452(1):1–17. https://doi.org/10.1042/BJ20121898

    Article  CAS  PubMed  Google Scholar 

  2. Oesch B, Westaway D, Walchli M, McKinley MP, Kent SB, Aebersold R, Barry RA, Tempst P et al (1985) A cellular gene encodes scrapie PrP 27-30 protein. Cell 40(4):735–746

    Article  CAS  PubMed  Google Scholar 

  3. Aguzzi A, Weissmann C (1997) Prion research: the next frontiers. Nature 389(6653):795–798. https://doi.org/10.1038/39758

    Article  CAS  PubMed  Google Scholar 

  4. Aguzzi A (2008) Staining, straining and restraining prions. Nat Neurosci 11(11):1239–1240. https://doi.org/10.1038/nn1108-1239

    Article  CAS  PubMed  Google Scholar 

  5. Sarnataro D, Pepe A, Zurzolo C (2017) Chapter three - cell biology of prion protein. In: Legname G, Vanni S (eds) Progress in molecular biology and translational science, vol 150. Academic Press, pp 57–82. doi:https://doi.org/10.1016/bs.pmbts.2017.06.018

    Google Scholar 

  6. Boellaard JW, Kao M, Schlote W, Diringer H (1991) Neuronal autophagy in experimental scrapie. Acta Neuropathol 82(3):225–228

    Article  CAS  PubMed  Google Scholar 

  7. Berridge MJ, Bootman MD, Lipp P (1998) Calcium--a life and death signal. Nature 395(6703):645–648. https://doi.org/10.1038/27094

    Article  CAS  PubMed  Google Scholar 

  8. Berridge MJ, Lipp P, Bootman MD (2000) Signal transduction. The calcium entry pas de deux. Science (New York, NY) 287(5458):1604–1605

    Article  CAS  Google Scholar 

  9. Paschen W (2003) Mechanisms of neuronal cell death: diverse roles of calcium in the various subcellular compartments. Cell Calcium 34(4–5):305–310

    Article  CAS  PubMed  Google Scholar 

  10. Florio T, Thellung S, Amico C, Robello M, Salmona M, Bugiani O, Tagliavini F, Forloni G et al (1998) Prion protein fragment 106-126 induces apoptotic cell death and impairment of L-type voltage-sensitive calcium channel activity in the GH3 cell line. J Neurosci Res 54(3):341–352. https://doi.org/10.1002/(sici)1097-4547(19981101)54:3<341::aid-jnr5>3.0.co;2-g

    Article  CAS  PubMed  Google Scholar 

  11. Thellung S, Florio T, Villa V, Corsaro A, Arena S, Amico C, Robello M, Salmona M et al (2000) Apoptotic cell death and impairment of L-type voltage-sensitive calcium channel activity in rat cerebellar granule cells treated with the prion protein fragment 106-126. Neurobiol Dis 7(4):299–309. https://doi.org/10.1006/nbdi.2000.0301

    Article  CAS  PubMed  Google Scholar 

  12. Mizushima N (2007) Autophagy: process and function. Genes Dev 21(22):2861–2873. https://doi.org/10.1101/gad.1599207

    Article  CAS  PubMed  Google Scholar 

  13. Kroemer G, Levine B (2008) Autophagic cell death: the story of a misnomer. Nat Rev Mol Cell Biol 9(12):1004–1010. https://doi.org/10.1038/nrm2529

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Klionsky DJ, Emr SD (2000) Autophagy as a regulated pathway of cellular degradation. Science (New York, NY) 290(5497):1717–1721

    Article  CAS  Google Scholar 

  15. Inoki K, Zhu T, Guan KL (2003) TSC2 mediates cellular energy response to control cell growth and survival. Cell 115(5):577–590

    Article  CAS  PubMed  Google Scholar 

  16. Corton JM, Gillespie JG, Hardie DG (1994) Role of the AMP-activated protein kinase in the cellular stress response. Curr Biol 4(4):315–324

    Article  CAS  PubMed  Google Scholar 

  17. Ng CH, Guan MS, Koh C, Ouyang X, Yu F, Tan EK, O'Neill SP, Zhang X et al (2012) AMP kinase activation mitigates dopaminergic dysfunction and mitochondrial abnormalities in Drosophila models of Parkinson’s disease. J Neurosci 32(41):14311–14317. https://doi.org/10.1523/jneurosci.0499-12.2012

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Hawley SA, Pan DA, Mustard KJ, Ross L, Bain J, Edelman AM, Frenguelli BG, Hardie DG (2005) Calmodulin-dependent protein kinase kinase-beta is an alternative upstream kinase for AMP-activated protein kinase. Cell Metab 2(1):9–19. https://doi.org/10.1016/j.cmet.2005.05.009

    Article  CAS  PubMed  Google Scholar 

  19. Shaw RJ, Bardeesy N, Manning BD, Lopez L, Kosmatka M, DePinho RA, Cantley LC (2004) The LKB1 tumor suppressor negatively regulates mTOR signaling. Cancer Cell 6(1):91–99. https://doi.org/10.1016/j.ccr.2004.06.007

    Article  CAS  PubMed  Google Scholar 

  20. Woods A, Dickerson K, Heath R, Hong SP, Momcilovic M, Johnstone SR, Carlson M, Carling D (2005) Ca2+/calmodulin-dependent protein kinase kinase-beta acts upstream of AMP-activated protein kinase in mammalian cells. Cell Metab 2(1):21–33. https://doi.org/10.1016/j.cmet.2005.06.005

    Article  CAS  PubMed  Google Scholar 

  21. Hoyer-Hansen M, Bastholm L, Szyniarowski P, Campanella M, Szabadkai G, Farkas T, Bianchi K, Fehrenbacher N et al (2007) Control of macroautophagy by calcium, calmodulin-dependent kinase kinase-beta, and Bcl-2. Mol Cell 25(2):193–205. https://doi.org/10.1016/j.molcel.2006.12.009

    Article  CAS  PubMed  Google Scholar 

  22. Godfraind T (2014) Calcium channel blockers in cardiovascular pharmacotherapy. J Cardiovasc Pharmacol Ther 19(6):501–515. https://doi.org/10.1177/1074248414530508

    Article  CAS  PubMed  Google Scholar 

  23. Meeting SFDRTA, Walker BC, Walker SR (1988) Trends and changes in drug research and development. Springer

  24. Patlak M (2004) From viper’s venom to drug design: treating hypertension. FASEB J 18(3):421

    Article  CAS  PubMed  Google Scholar 

  25. Alvin Z, Laurence GG, Coleman BR, Zhao A, Hajj-Moussa M, Haddad GE (2011) Regulation of L-type inward calcium channel activity by captopril and angiotensin II via the phosphatidyl inositol 3-kinase pathway in cardiomyocytes from volume-overload hypertrophied rat hearts. Can J Physiol Pharmacol 89(3):206–215. https://doi.org/10.1139/y11-011

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Tsien RY, Pozzan T, Rink TJ (1982) T-cell mitogens cause early changes in cytoplasmic free Ca2+ and membrane potential in lymphocytes. Nature 295(5844):68–71

    Article  CAS  PubMed  Google Scholar 

  27. Florio T, Grimaldi M, Scorziello A, Salmona M, Bugiani O, Tagliavini F, Forloni G, Schettini G (1996) Intracellular calcium rise through L-type calcium channels, as molecular mechanism for prion protein fragment 106-126-induced astroglial proliferation. Biochem Biophys Res Commun 228(2):397–405. https://doi.org/10.1006/bbrc.1996.1673

    Article  CAS  PubMed  Google Scholar 

  28. Herms JW, Madlung A, Brown DR, Kretzschmar HA (1997) Increase of intracellular free Ca2+ in microglia activated by prion protein fragment. Glia 21(2):253–257

    Article  CAS  PubMed  Google Scholar 

  29. Korotzer AR, Whittemore ER, Cotman CW (1995) Differential regulation by beta-amyloid peptides of intracellular free Ca2+ concentration in cultured rat microglia. Eur J Pharmacol 288(2):125–130

    Article  CAS  PubMed  Google Scholar 

  30. Silei V, Fabrizi C, Venturini G, Salmona M, Bugiani O, Tagliavini F, Lauro GM (1999) Activation of microglial cells by PrP and beta-amyloid fragments raises intracellular calcium through L-type voltage sensitive calcium channels. Brain Res 818(1):168–170

    Article  CAS  PubMed  Google Scholar 

  31. Rubinsztein DC, Cuervo AM, Ravikumar B, Sarkar S, Korolchuk V, Kaushik S, Klionsky DJ (2009) In search of an “autophagomometer”. Autophagy 5(5):585–589

    Article  CAS  PubMed  Google Scholar 

  32. Bjorkoy G, Lamark T, Brech A, Outzen H, Perander M, Overvatn A, Stenmark H, Johansen T (2005) p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol 171(4):603–614. https://doi.org/10.1083/jcb.200507002

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Brouwer RM, Bolli P, Erné P, Conen D, Kiowski W, Bühler FR (1985) Antihypertensive treatment using calcium antagonists in combination with captopril rather than diuretics. J Cardiovasc Pharmacol 7(1):S88–S91

    Article  PubMed  Google Scholar 

  34. Asaad MM, Antonaccio MJ (1982) Vascular wall renin in spontaneously hypertensive rats. Potential relevance to hypertension maintenance and antihypertensive effect of captopril. Hypertension 4(4):487–493. https://doi.org/10.1161/01.hyp.4.4.487

    Article  CAS  PubMed  Google Scholar 

  35. Schoenberger JA, Testa M, Ross AD, Brennan WK, Bannon JA (1990) Efficacy, safety, and quality-of-life assessment of captopril antihypertensive therapy in clinical practice. Arch Intern Med 150(2):301–306. https://doi.org/10.1001/archinte.1990.00390140051011

    Article  CAS  PubMed  Google Scholar 

  36. AbdAlla S, Langer A, Fu X, Quitterer U (2013) ACE inhibition with captopril retards the development of signs of neurodegeneration in an animal model of Alzheimer’s disease. Int J Mol Sci 14(8):16917–16942. https://doi.org/10.3390/ijms140816917

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Connell BJ, Khan BV, Rajagopal D, Saleh TM (2012) Novel neurovascular protective agents: effects of INV-155, INV-157, INV-159, and INV-161 versus lipoic acid and captopril in a rat stroke model. Cardiol Res Pract 2012:319230. https://doi.org/10.1155/2012/319230

    Article  PubMed  PubMed Central  Google Scholar 

  38. Sengul G, Coskun S, Cakir M, Coban MK, Saruhan F, Hacimuftuoglu A (2011) Neuroprotective effect of ACE inhibitors in glutamate - induced neurotoxicity: rat neuron culture study. Turk Neurosurg 21(3):367–371. https://doi.org/10.5137/1019-5149.jtn.4313-11.0

    Article  PubMed  Google Scholar 

  39. Sonsalla PK, Coleman C, Wong LY, Harris SL, Richardson JR, Gadad BS, Li W, German DC (2013) The angiotensin converting enzyme inhibitor captopril protects nigrostriatal dopamine neurons in animal models of parkinsonism. Exp Neurol 250:376–383. https://doi.org/10.1016/j.expneurol.2013.10.014

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Inoki K, Kim J, Guan KL (2012) AMPK and mTOR in cellular energy homeostasis and drug targets. Annu Rev Pharmacol Toxicol 52:381–400. https://doi.org/10.1146/annurev-pharmtox-010611-134537

    Article  CAS  PubMed  Google Scholar 

  41. Otomo C, Metlagel Z, Takaesu G, Otomo T (2013) Structure of the human ATG12~ATG5 conjugate required for LC3 lipidation in autophagy. Nat Struct Mol Biol 20(1):59–66. https://doi.org/10.1038/nsmb.2431

    Article  CAS  PubMed  Google Scholar 

  42. Yang YP, Liang ZQ, Gu ZL, Qin ZH (2005) Molecular mechanism and regulation of autophagy. Acta Pharmacol Sin 26(12):1421–1434. https://doi.org/10.1111/j.1745-7254.2005.00235.x

    Article  CAS  PubMed  Google Scholar 

  43. Levine B, Yuan J (2005) Autophagy in cell death: an innocent convict? J Clin Invest 115(10):2679–2688. https://doi.org/10.1172/jci26390

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Moscat J, Diaz-Meco MT (2009) p62 at the crossroads of autophagy, apoptosis, and cancer. Cell 137(6):1001–1004. https://doi.org/10.1016/j.cell.2009.05.023

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Mizushima N, Yoshimori T (2007) How to interpret LC3 immunoblotting. Autophagy 3(6):542–545

    Article  CAS  PubMed  Google Scholar 

  46. Bauvy C, Meijer AJ, Codogno P (2009) Assaying of autophagic protein degradation. Methods Enzymol 452:47–61. https://doi.org/10.1016/s0076-6879(08)03604-5

    Article  CAS  PubMed  Google Scholar 

  47. Yoon YH, Cho KS, Hwang JJ, Lee SJ, Choi JA, Koh JY (2010) Induction of lysosomal dilatation, arrested autophagy, and cell death by chloroquine in cultured ARPE-19 cells. Invest Ophthalmol Vis Sci 51(11):6030–6037. https://doi.org/10.1167/iovs.10-5278

    Article  PubMed  Google Scholar 

  48. Saiki S, Sasazawa Y, Imamichi Y, Kawajiri S, Fujimaki T, Tanida I, Kobayashi H, Sato F et al (2011) Caffeine induces apoptosis by enhancement of autophagy via PI3K/Akt/mTOR/p70S6K inhibition. Autophagy 7(2):176–187

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Kumar D, Shankar S, Srivastava RK (2013) Rottlerin-induced autophagy leads to the apoptosis in breast cancer stem cells: molecular mechanisms. Mol Cancer 12(1):171. https://doi.org/10.1186/1476-4598-12-171

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Arsikin K, Kravic-Stevovic T, Jovanovic M, Ristic B, Tovilovic G, Zogovic N, Bumbasirevic V, Trajkovic V et al (2012) Autophagy-dependent and -independent involvement of AMP-activated protein kinase in 6-hydroxydopamine toxicity to SH-SY5Y neuroblastoma cells. Biochim Biophys Acta 1822(11):1826–1836. https://doi.org/10.1016/j.bbadis.2012.08.006

    Article  CAS  PubMed  Google Scholar 

  51. Jeong JK, Moon MH, Bae BC, Lee YJ, Seol JW, Kang HS, Kim JS, Kang SJ et al (2012) Autophagy induced by resveratrol prevents human prion protein-mediated neurotoxicity. Neurosci Res 73(2):99–105. https://doi.org/10.1016/j.neures.2012.03.005

    Article  CAS  PubMed  Google Scholar 

  52. Lee JH, Jeong JK, Park SY (2014) Sulforaphane-induced autophagy flux prevents prion protein-mediated neurotoxicity through AMPK pathway. Neuroscience 278:31–39. https://doi.org/10.1016/j.neuroscience.2014.07.072

    Article  CAS  PubMed  Google Scholar 

  53. Uchiyama Y, Koike M, Shibata M (2008) Autophagic neuron death in neonatal brain ischemia/hypoxia. Autophagy 4(4):404–408

    Article  CAS  PubMed  Google Scholar 

  54. Shintani T, Klionsky DJ (2004) Autophagy in health and disease: a double-edged sword. Science (New York, NY) 306(5698):990–995. https://doi.org/10.1126/science.1099993

    Article  CAS  Google Scholar 

  55. Deretic V, Saitoh T, Akira S (2013) Autophagy in infection, inflammation and immunity. Nat Rev Immunol 13(10):722–737. https://doi.org/10.1038/nri3532

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Wong E, Cuervo AM (2010) Autophagy gone awry in neurodegenerative diseases. Nat Neurosci 13(7):805–811. https://doi.org/10.1038/nn.2575

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This study was supported by the National Research Foundation of the Korea Grant (MISP) funded by the Korean Government (2016R1A2B2009293).

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  1. Ji-Hong Moon and Jae-Kyo Jeong contributed equally to this work.

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  1. Biosafety Research Institute, College of Veterinary Medicine, Chonbuk National University, Iksan, Jeonbuk, 54596, South Korea

    Ji-Hong Moon, Jae-Kyo Jeong, Jeong-Min Hong, Jae-Won Seol & Sang-Youel Park

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  1. Ji-Hong Moon
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Contributions

J.H.M., J.K.J., J.W.S., and S.Y.P. designed, executed the study, analyzed data, and wrote the manuscript. J.H.M. and J.K.J performed the electron microscopy. All authors have read and approved the final manuscript.

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Correspondence to Sang-Youel Park.

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Ethical approval for the project was granted by the institutional review board of the Chonbuk National University.

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Moon, JH., Jeong, JK., Hong, JM. et al. Inhibition of Autophagy by Captopril Attenuates Prion Peptide-Mediated Neuronal Apoptosis via AMPK Activation. Mol Neurobiol 56, 4192–4202 (2019). https://doi.org/10.1007/s12035-018-1370-8

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