Advertisement

Apoptosis

, Volume 21, Issue 11, pp 1227–1239 | Cite as

Endoplasmic reticulum (ER) stress triggers Hax1-dependent mitochondrial apoptotic events in cardiac cells

  • Eltyeb AbdelwahidEmail author
  • Haijie Li
  • Jianxin Wu
  • Ana Carolina Irioda
  • Katherine Athayde Teixeira de Carvalho
  • Xuelai LuoEmail author
Article

Abstract

Cardiomyocyte apoptosis is a major process in pathogenesis of a number of heart diseases, including ischemic heart diseases and cardiac failure. Ensuring survival of cardiac cells by blocking apoptotic events is an important strategy to improve cardiac function. Although the role of ER disruption in inducing apoptosis has been demonstrated, we do not yet fully understand how it influences the mitochondrial apoptotic machinery in cardiac cell models. Recent investigations have provided evidences that the prosurvival protein HCLS1-associated protein X-1 (Hax1) protein is intimately associated with the pathogenesis of heart disease, mitochondrial biology, and protection from apoptotic cell death. To study the role of Hax1 upon ER stress induction, Hax1 was overexpressed in cardiac cells subjected to ER stress, and cell death parameters as well as mitochondrial alterations were examined. Our results demonstrated that the Hax1 is significantly downregulated in cardiac cells upon ER stress induction. Moreover, overexpression of Hax1 protected from apoptotic events triggered by Tunicamycin-induced ER stress. Upon treatment with Tunicamycin, Hax1 protected from mitochondrial fission, downregulation of mitofusins 1 and 2 (MFN1 and MFN2), loss of mitochondrial membrane potential (∆Ψm), production of reactive oxygen species (ROS) and apoptotic cell death. Taken together, our results suggest that Hax1 inhibits ER stress-induced apoptosis at both the pre- and post-mitochondrial levels. These findings may offer an opportunity to develop new agents that inhibit cell death in the diseased heart.

Keywords

Hax1 ER stress Mitochondria Mitofusin Mitochondrial fission ROS Apoptosis 

Notes

Acknowledgments

We thank Dr. Luca Pellegrini (Faculty of Medicine, Université Laval, Quebec, QC, Canada) for Hax1 overexpression construct, Dr. Thomas Simmen for discussion and Dr. Aikaterini Kontrogianni-Konstantopoulos for kindly sharing information on Hax1 expression. We are grateful to Mr. Denislam Zaripov for art drawing. E.A. was supported by the National Heart, Lung, and Blood Institute (NIH/NHLBI), Grant SP0012613. X.L. was supported by the National Natural Science Foundation of China (81272278). K.A.T.C was supported by Coordination for the Improvement of Higher Education Personnel (CAPES) of Brazil, Grant PE 1711.

References

  1. 1.
    van Empel VP, Bertrand AT, Hofstra L, Crijns HJ, Doevendans PA, De Windt LJ (2005) Myocyte apoptosis in heart failure. Cardiovasc Res 67:21–29CrossRefPubMedGoogle Scholar
  2. 2.
    Wang Y, Huang S, Sah VP, Ross J Jr, Brown JH, Han J et al (1998) Cardiac muscle cell hypertrophy and apoptosis induced by distinct members of the p38 mitogen-activated protein kinase family. J Biol Chem 273:2161–2168CrossRefPubMedGoogle Scholar
  3. 3.
    Mel’nikova NP, Timoshin SS, Jivotova EY, Pelliniemi LJ, Jokinen E, Abdelwahid E (2006) Angiotensin-II activates apoptosis, proliferation and protein synthesis in the left heart ventricle of newborn albino rats. Int J Cardiol 112:219–222CrossRefPubMedGoogle Scholar
  4. 4.
    Abdelwahid E, Smith G (2007) Apoptosis in chronic heart failure. Int J Cardiol 114:375CrossRefPubMedGoogle Scholar
  5. 5.
    Abdelwahid E, Petrovic D, Feng Q, Mistiaen WP (2007) Molecular mechanisms and new developments in the regulation of programmed cell death (apoptosis) and its role in pathogenesis of heart diseases. Nova Science Publishers, Inc, New YorkGoogle Scholar
  6. 6.
    Abdelwahid E, Rolland S, Teng X, Conradt B, Hardwick JM, White K (2011) Mitochondrial involvement in cell death of non-mammalian eukaryotes. Biochim Biophys Acta 1813:597–607CrossRefPubMedGoogle Scholar
  7. 7.
    Abdelwahid E, Yokokura T, Krieser RJ, Balasundaram S, Fowle WH, White K (2007) Mitochondrial disruption in Drosophila apoptosis. Dev Cell 12:793–806CrossRefPubMedGoogle Scholar
  8. 8.
    Yu T, Sheu SS, Robotham JL, Yoon Y (2008) Mitochondrial fission mediates high glucose-induced cell death through elevated production of reactive oxygen species. Cardiovasc Res 79:341–351CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Youle RJ, Karbowski M (2005) Mitochondrial fission in apoptosis. Nat Rev Mol Cell Biol 6:657–663CrossRefPubMedGoogle Scholar
  10. 10.
    Su B, Wang X, Bonda D, Perry G, Smith M, Zhu X (2010) Abnormal mitochondrial dynamics–a novel therapeutic target for Alzheimer’s disease? Mol Neurobiol 41:87–96CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Kaufman RJ (2002) Orchestrating the unfolded protein response in health and disease. J Clin Invest 110:1389–1398CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Ruddock LW, Molinari M (2006) N-glycan processing in ER quality control. J Cell Sci 119:4373–4380CrossRefPubMedGoogle Scholar
  13. 13.
    Ono Y, Shimazawa M, Ishisaka M, Oyagi A, Tsuruma K, Hara H (2012) Imipramine protects mouse hippocampus against Tunicamycin-induced cell death. Eur J Pharmacol 696:83–88CrossRefPubMedGoogle Scholar
  14. 14.
    Gorlach A, Klappa P, Kietzmann T (2006) The endoplasmic reticulum: folding, calcium homeostasis, signaling, and redox control. Antioxid Redox Signal 8:1391–1418CrossRefPubMedGoogle Scholar
  15. 15.
    Sidrauski C, Chapman R, Walter P (1998) The unfolded protein response: an intracellular signalling pathway with many surprising features. Trends Cell Biol 8:245–249CrossRefPubMedGoogle Scholar
  16. 16.
    Urano F, Bertolotti A, Ron D (2000) IRE1 and efferent signaling from the endoplasmic reticulum. J Cell Sci 113(Pt 21):3697–3702PubMedGoogle Scholar
  17. 17.
    Nakagawa T, Zhu H, Morishima N, Li E, Xu J, Yankner BA et al (2000) Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta. Nature 403:98–103CrossRefPubMedGoogle Scholar
  18. 18.
    Puthalakath H, O’Reilly LA, Gunn P, Lee L, Kelly PN, Huntington ND et al (2007) ER stress triggers apoptosis by activating BH3-only protein Bim. Cell 129:1337–1349CrossRefPubMedGoogle Scholar
  19. 19.
    Kornmann B, Walter P (2010) ERMES-mediated ER-mitochondria contacts: molecular hubs for the regulation of mitochondrial biology. J Cell Sci 123:1389–1393CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Pizzo P, Pozzan T (2007) Mitochondria-endoplasmic reticulum choreography: structure and signaling dynamics. Trends Cell Biol 17:511–517CrossRefPubMedGoogle Scholar
  21. 21.
    Csordas G, Varnai P, Golenar T, Roy S, Purkins G, Schneider TG et al (2010) Imaging interorganelle contacts and local calcium dynamics at the ER-mitochondrial interface. Mol Cell 39:121–132CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Pirot P, Ortis F, Cnop M, Ma Y, Hendershot LM, Eizirik DL et al (2007) Transcriptional regulation of the endoplasmic reticulum stress gene chop in pancreatic insulin-producing cells. Diabetes 56:1069–1077CrossRefPubMedGoogle Scholar
  23. 23.
    Suzuki Y, Demoliere C, Kitamura D, Takeshita H, Deuschle U, Watanabe T (1997) HAX-1, a novel intracellular protein, localized on mitochondria, directly associates with HS1, a substrate of Src family tyrosine kinases. J Immunol 158:2736–2744PubMedGoogle Scholar
  24. 24.
    Lam CK, Zhao W, Cai W, Vafiadaki E, Florea SM, Ren X et al (2013) Novel role of HAX-1 in ischemic injury protection involvement of heat shock protein 90. Circ Res 112:79–89CrossRefPubMedGoogle Scholar
  25. 25.
    Han Y, Chen YS, Liu Z, Bodyak N, Rigor D, Bisping E et al (2006) Overexpression of HAX-1 protects cardiac myocytes from apoptosis through caspase-9 inhibition. Circ Res 99:415–423CrossRefPubMedGoogle Scholar
  26. 26.
    Chao JR, Parganas E, Boyd K, Hong CY, Opferman JT, Ihle JN (2008) Hax1-mediated processing of HtrA2 by Parl allows survival of lymphocytes and neurons. Nature 452:98–102CrossRefPubMedGoogle Scholar
  27. 27.
    Fadeel B, Grzybowska E (2009) HAX-1: a multifunctional protein with emerging roles in human disease. Biochim Biophys Acta 1790:1139–1148CrossRefPubMedGoogle Scholar
  28. 28.
    Boztug K, Ding XQ, Hartmann H, Ziesenitz L, Schaffer AA, Diestelhorst J et al (2010) HAX1 mutations causing severe congenital neuropenia and neurological disease lead to cerebral microstructural abnormalities documented by quantitative MRI. Am J Med Genet A 152 A:3157–3163CrossRefGoogle Scholar
  29. 29.
    Lanciotti M, Indaco S, Bonanomi S, Coliva T, Mastrodicasa E, Caridi G et al (2010) Novel HAX1 gene mutations associated to neurodevelopment abnormalities in two Italian patients with severe congenital neutropenia. Haematologica 95:168–169CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Koontz J, Kontrogianni-Konstantopoulos A (2014) Competition through dimerization between antiapoptotic and proapoptotic HS-1-associated protein X-1 (Hax-1). J Biol Chem 289:3468–3477CrossRefPubMedGoogle Scholar
  31. 31.
    Zhao W, Waggoner JR, Zhang ZG, Lam CK, Han P, Qian J et al (2009) The anti-apoptotic protein HAX-1 is a regulator of cardiac function. Proc Natl Acad Sci USA 106:20776–20781CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Grzybowska EA, Sarnowska E, Konopinski R, Wilczynska A, Sarnowski TJ, Siedlecki JA (2006) Identification and expression analysis of alternative splice variants of the rat Hax-1 gene. Gene 371:84–92CrossRefPubMedGoogle Scholar
  33. 33.
    Jeyaraju DV, Cisbani G, De Brito OM, Koonin EV, Pellegrini L (2009) Hax1 lacks BH modules and is peripherally associated to heavy membranes: implications for Omi/HtrA2 and PARL activity in the regulation of mitochondrial stress and apoptosis. Cell Death Differ 16:1622–1629CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Gurgen D, Hegner B, Kusch A, Catar R, Chaykovska L, Hoff U et al (2011) Estrogen receptor-beta signals left ventricular hypertrophy sex differences in normotensive deoxycorticosterone acetate-salt mice. Hypertension 57:648–654CrossRefPubMedGoogle Scholar
  35. 35.
    Santra M, Skorski T, Calabretta B, Lattime EC, Iozzo RV (1995) De novo decorin gene expression suppresses the malignant phenotype in human colon cancer cells. Proc Natl Acad Sci USA 92:7016–7020CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Frank S, Gaume B, Bergmann-Leitner ES, Leitner WW, Robert EG, Catez F et al (2001) The role of dynamin-related protein 1, a mediator of mitochondrial fission, in apoptosis. Dev Cell 1:515–525CrossRefPubMedGoogle Scholar
  37. 37.
    Liot G, Bossy B, Lubitz S, Kushnareva Y, Sejbuk N, Bossy-Wetzel E (2009) Complex II inhibition by 3-NP causes mitochondrial fragmentation and neuronal cell death via an NMDA- and ROS-dependent pathway. Cell Death Differ 16:899–909CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Chou CH, Lin CC, Yang MC, Wei CC, Liao HD, Lin RC et al (2012) GSK3beta-mediated Drp1 phosphorylation induced elongated mitochondrial morphology against oxidative stress. PLoS One 7:e49112CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Eura Y, Ishihara N, Yokota S, Mihara K (2003) Two mitofusin proteins, mammalian homologues of FZO, with distinct functions are both required for mitochondrial fusion. J Biochem 134:333–344CrossRefPubMedGoogle Scholar
  40. 40.
    Yaglom JA, Ekhterae D, Gabai VL, Sherman MY (2003) Regulation of necrosis of H9c2 myogenic cells upon transient energy deprivation. Rapid deenergization of mitochondria precedes necrosis and is controlled by reactive oxygen species, stress kinase JNK, HSP72 and ARC. J Biol Chem 278:50483–50496CrossRefPubMedGoogle Scholar
  41. 41.
    Shen M, Wang L, Wang B, Wang T, Yang G, Shen L et al (2014) Activation of volume-sensitive outwardly rectifying chloride channel by ROS contributes to ER stress and cardiac contractile dysfunction: involvement of CHOP through Wnt. Cell Death Dis 5:e1528CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Li G, Scull C, Ozcan L, Tabas I (2010) NADPH oxidase links endoplasmic reticulum stress, oxidative stress, and PKR activation to induce apoptosis. J Cell Biol 191:1113–1125CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Tanjore H, Lawson WE, Blackwell TS (2013) Endoplasmic reticulum stress as a pro-fibrotic stimulus. Biochim Biophys Acta 1832:940–947CrossRefPubMedGoogle Scholar
  44. 44.
    Han C, Nam MK, Park HJ, Seong YM, Kang S, Rhim H (2008) Tunicamycin-induced ER stress upregulates the expression of mitochondrial HtrA2 and promotes apoptosis through the cytosolic release of HtrA2. J Microbiol Biotechnol 18:1197–1202PubMedGoogle Scholar
  45. 45.
    Ngoh GA, Papanicolaou KN, Walsh K (2012) Loss of mitofusin 2 promotes endoplasmic reticulum stress. J Biol Chem 287:20321–20332CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Korobova F, Ramabhadran V, Higgs HN (2013) An actin-dependent step in mitochondrial fission mediated by the ER-associated formin INF2. Science 339:464–467CrossRefPubMedGoogle Scholar
  47. 47.
    Friedman JR, Lackner LL, West M, DiBenedetto JR, Nunnari J, Voeltz GK (2011) ER tubules mark sites of mitochondrial division. Science 334:358–362CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Iwasawa R, Mahul-Mellier AL, Datler C, Pazarentzos E, Grimm S (2011) Fis1 and Bap31 bridge the mitochondria-ER interface to establish a platform for apoptosis induction. EMBO J 30:556–568CrossRefPubMedGoogle Scholar
  49. 49.
    Ong SB, Subrayan S, Lim SY, Yellon DM, Davidson SM, Hausenloy DJ (2010) Inhibiting mitochondrial fission protects the heart against ischemia/reperfusion injury. Circulation 121:2012–2022CrossRefPubMedGoogle Scholar
  50. 50.
    Grossmann J, Walther K, Artinger M, Kiessling S, Scholmerich J (2001) Apoptotic signaling during initiation of detachment-induced apoptosis (“anoikis”) of primary human intestinal epithelial cells. Cell Growth Differ 12:147–155PubMedGoogle Scholar
  51. 51.
    Grossmann J (2002) Molecular mechanisms of “detachment-induced apoptosis–Anoikis”. Apoptosis 7:247–260CrossRefPubMedGoogle Scholar
  52. 52.
    Sun J, Sun G, Meng X, Wang H, Wang M, Qin M et al (2013) Ginsenoside RK3 Prevents Hypoxia-Reoxygenation Induced Apoptosis in H9c2 Cardiomyocytes via AKT and MAPK Pathway. Evid Based Complement Alternat Med 2013:690190PubMedPubMedCentralGoogle Scholar
  53. 53.
    Abdelwahid E, Kalvelyte A, Stulpinas A, de Carvalho KA, Guarita-Souza LC, Foldes G (2016) Stem cell death and survival in heart regeneration and repair. Apoptosis 21:252–268CrossRefPubMedGoogle Scholar
  54. 54.
    Abdelwahid E, Rice D, Pelliniemi LJ, Jokinen E (2001) Overlapping and differential localization of Bmp-2, Bmp-4, Msx-2 and apoptosis in the endocardial cushion and adjacent tissues of the developing mouse heart. Cell Tissue Res 305:67–78CrossRefPubMedGoogle Scholar
  55. 55.
    Wang C, Li YZ, Wang XR, Lu ZR, Shi DZ, Liu XH (2012) Panax quinquefolium saponins reduce myocardial hypoxia-reoxygenation injury by inhibiting excessive endoplasmic reticulum stress. Shock 37:228–233CrossRefPubMedGoogle Scholar
  56. 56.
    Wu XD, Zhang ZY, Sun S, Li YZ, Wang XR, Zhu XQ et al (2013) Hypoxic preconditioning protects microvascular endothelial cells against hypoxia/reoxygenation injury by attenuating endoplasmic reticulum stress. Apoptosis 18:85–98CrossRefPubMedGoogle Scholar
  57. 57.
    Lam CK, Zhao W, Liu GS, Cai WF, Gardner G, Adly G et al (2015) HAX-1 regulates cyclophilin-D levels and mitochondria permeability transition pore in the heart. Proc Natl Acad Sci USA 112:E6466–E6475CrossRefPubMedPubMedCentralGoogle Scholar
  58. 58.
    Lam AK, Galione A, Lai FA, Zissimopoulos S (2013) Hax-1 identified as a two-pore channel (TPC)-binding protein. FEBS Lett 587:3782–3786CrossRefPubMedGoogle Scholar
  59. 59.
    Radhika V, Onesime D, Ha JH, Dhanasekaran N (2004) Galpha13 stimulates cell migration through cortactin-interacting protein Hax-1. J Biol Chem 279:49406–49413CrossRefPubMedGoogle Scholar
  60. 60.
    Kawaguchi Y, Nakajima K, Igarashi M, Morita T, Tanaka M, Suzuki M et al (2000) Interaction of Epstein-Barr virus nuclear antigen leader protein (EBNA-LP) with HS1-associated protein X-1: implication of cytoplasmic function of EBNA-LP. J Virol 74:10104–10111CrossRefPubMedPubMedCentralGoogle Scholar
  61. 61.
    Vasquez-Trincado C, Garcia-Carvajal I, Pennanen C, Parra V, Hill JA, Rothermel BA et al (2016) Mitochondrial dynamics, mitophagy and cardiovascular disease. J Physiol 594:509–525CrossRefPubMedGoogle Scholar
  62. 62.
    Kane LA, Youle RJ (2010) Mitochondrial fission and fusion and their roles in the heart. J Mol Med (Berl) 88:971–979CrossRefGoogle Scholar
  63. 63.
    Hom J, Sheu SS (2009) Morphological dynamics of mitochondria–a special emphasis on cardiac muscle cells. J Mol Cell Cardiol 46:811–820CrossRefPubMedPubMedCentralGoogle Scholar
  64. 64.
    Ong SB, Hausenloy DJ (2010) Mitochondrial morphology and cardiovascular disease. Cardiovasc Res 88:16–29CrossRefPubMedPubMedCentralGoogle Scholar
  65. 65.
    Chen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC (2003) Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. J Cell Biol 160:189–200CrossRefPubMedPubMedCentralGoogle Scholar
  66. 66.
    Papanicolaou KN, Ngoh GA, Dabkowski ER, O’Connell KA, Ribeiro RF Jr, Stanley WC et al (2012) Cardiomyocyte deletion of mitofusin-1 leads to mitochondrial fragmentation and improves tolerance to ROS-induced mitochondrial dysfunction and cell death. Am J Physiol Heart Circ Physiol 302:H167–H179CrossRefPubMedGoogle Scholar
  67. 67.
    Chen Y, Liu Y, Dorn GW 2nd (2011) Mitochondrial fusion is essential for organelle function and cardiac homeostasis. Circ Res 109:1327–1331CrossRefPubMedPubMedCentralGoogle Scholar
  68. 68.
    Skulachev VP (2001) Mitochondrial filaments and clusters as intracellular power-transmitting cables. Trends Biochem Sci 26:23–29CrossRefPubMedGoogle Scholar
  69. 69.
    Narula J, Pandey P, Arbustini E, Haider N, Narula N, Kolodgie FD et al (1999) Apoptosis in heart failure: release of cytochrome c from mitochondria and activation of caspase-3 in human cardiomyopathy. Proc Natl Acad Sci USA 96:8144–8149CrossRefPubMedPubMedCentralGoogle Scholar
  70. 70.
    Papanicolaou KN, Khairallah RJ, Ngoh GA, Chikando A, Luptak I, O’Shea KM et al (2011) Mitofusin-2 maintains mitochondrial structure and contributes to stress-induced permeability transition in cardiac myocytes. Mol Cell Biol 31:1309–1328CrossRefPubMedPubMedCentralGoogle Scholar
  71. 71.
    Shi Y (2002) Mechanisms of caspase activation and inhibition during apoptosis. Mol Cell 9:459–470CrossRefPubMedGoogle Scholar
  72. 72.
    Olsson M, Zhivotovsky B (2011) Caspases and cancer. Cell Death Differ 18:1441–1449CrossRefPubMedPubMedCentralGoogle Scholar
  73. 73.
    Guo Y, Srinivasula SM, Druilhe A, Fernandes-Alnemri T, Alnemri ES (2002) Caspase-2 induces apoptosis by releasing proapoptotic proteins from mitochondria. J Biol Chem 277:13430–13437CrossRefPubMedGoogle Scholar
  74. 74.
    Enoksson M, Robertson JD, Gogvadze V, Bu P, Kropotov A, Zhivotovsky B et al (2004) Caspase-2 permeabilizes the outer mitochondrial membrane and disrupts the binding of cytochrome c to anionic phospholipids. J Biol Chem 279:49575–49578CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Eltyeb Abdelwahid
    • 1
    Email author
  • Haijie Li
    • 2
  • Jianxin Wu
    • 2
  • Ana Carolina Irioda
    • 3
  • Katherine Athayde Teixeira de Carvalho
    • 3
  • Xuelai Luo
    • 2
    Email author
  1. 1.Feinberg Cardiovascular Research Institute, Feinberg School of MedicineNorthwestern UniversityChicagoUSA
  2. 2.Cancer Research Institute, Tongji Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  3. 3.Pequeno Príncipe Faculty, Cell Therapy and Biotechnology in Regenerative Medicine Department, The Pelé Pequeno Príncipe InstituteChild and Adolescent Health ResearchCuritibaBrazil

Personalised recommendations