Abstract
Ebola virus (EBOV) causes a highly lethal hemorrhagic fever syndrome in humans and has been associated with mortality rates of up to 91% in Zaire, the most lethal strain. Though the viral envelope glycoprotein (GP) mediates widespread inflammation and cellular damage, these changes have mainly focused on alterations at the protein level, the role of microRNAs (miRNAs) in the molecular pathogenesis underlying this lethal disease is not fully understood. Here, we report that the mi-RNAs hsa-miR-1246, hsa-miR-320a and hsa-miR-196b-5p were induced in human umbilical vein endothelial cells (HUVECs) following expression of EBOV GP. Among the proteins encoded by predicted targets of these miRNAs, the adhesion-related molecules tissue factor pathway inhibitor (TFPI), dystroglycan1 (DAG1) and the caspase 8 and FADD-like apoptosis regulator (CFLAR) were significantly downregulated in EBOV GP-expressing HUVECs. Moreover, inhibition of hsa-miR-1246, hsa-miR-320a and hsa-miR-196b-5p, or overexpression of TFPI, DAG1 and CFLAR rescued the cell viability that was induced by EBOV GP. Our results provide a novel molecular basis for EBOV pathogenesis and may contribute to the development of strategies to protect against future EBOV pandemics.
Article PDF
References
Colebunders R, Borchert M. Ebola haemorrhagic fever-a review. J Infect, 2000, 40: 16–20
Kuhn JH, Becker S, Ebihara H, Geisbert TW, Johnson KM, Kawaoka Y, Lipkin WI, Negredo AI, Netesov SV, Nichol ST, Palacios G, Peters CJ, Tenorio A, Volchkov VE, Jahrling PB. Proposal for a revised taxonomy of the family filoviridae: classification, names of taxa and viruses, and virus abbreviations. Arch Virol, 2010, 155: 2083–2103
Nkoghe D, Formenty P, Leroy EM, Nnegue S, Edou SY, Ba JI, Allarangar Y, Cabore J, Bachy C, Andraghetti R, de Benoist AC, Galanis E, Rose A, Bausch D, Reynolds M, Rollin P, Choueibou C, Shongo R, Gergonne B, Kone LM, Yada A, Roth C, Mve MT. Multiple Ebola virus haemorrhagic fever outbreaks in gabon, from October 2001 to April 2002. Bull Soc Pathol Exot, 2005, 98: 224–229
Simmons G, Wool-Lewis RJ, Baribaud F, Netter RC, Bates P. Ebola virus glycoproteins induce global surface protein down-modulation and loss of cell adherence. J Virol, 2002, 76: 2518–2528
Peters CJ, LeDuc JW. An introduction to Ebola: the virus and the disease. J Infect Dis, 1999, 179(Suppl 1): ix–xvi
Takada A, Watanabe S, Ito H, Okazaki K, Kida H, Kawaoka Y. Downregulation of beta1 integrins by Ebola virus glycoprotein: implication for virus entry. Virology, 2000, 278: 20–26
Chan SY, Empig CJ, Welte FJ, Speck RF, Schmaljohn A, Kreisberg JF, Goldsmith MA. Folate receptor-alpha is a cofactor for cellular entry by marburg and Ebola viruses. Cell, 2001, 106: 117–126
Alvarez CP, Lasala F, Carrillo J, Muniz O, Corbi AL, Delgado R. C-type lectins DC-SIGN and L-SIGN mediate cellular entry by Ebola virus in cis and in trans. J Virol, 2002, 76: 6841–6844
Takada A, Fujioka K, Tsuiji M, Morikawa A, Higashi N, Ebihara H, Kobasa D, Feldmann H, Irimura T, Kawaoka Y. Human macrophage C-type lectin specific for galactose and N-acetylgalactosamine promotes filovirus entry. J Virol, 2004, 78: 2943–2947
Shimojima M, Takada A, Ebihara H, Neumann G, Fujioka K, Irimura T, Jones S, Feldmann H, Kawaoka Y. Tyro3 family-mediated cell entry of Ebola and Marburg viruses. J Virol, 2006, 80: 10109–10116
Carette JE, Raaben M, Wong AC, Herbert AS, Obernosterer G, Mulherkar N, Kuehne AI, Kranzusch PJ, Griffin AM, Ruthel G, Dal Cin P, Dye JM, Whelan SP, Chandran K, Brummelkamp TR. Ebola virus entry requires the cholesterol transporter Niemann-Pick C1. Nature, 2011, 477: 340–343
Kondratowicz AS, Lennemann NJ, Sinn PL, Davey RA, Hunt CL, Moller-Tank S, Meyerholz DK, Rennert P, Mullins RF, Brindley M, Sandersfeld LM, Quinn K, Weller M, McCray PB, Jr., Chiorini J, Maury W. T-cell immunoglobulin and mucin domain 1 (TIM-1) is a receptor for Zaire Ebolavirus and Lake Victoria Marburgvirus. Proc Natl Acad Sci USA, 2011, 108: 8426–8431
Miller EH, Obernosterer G, Raaben M, Herbert AS, Deffieu MS, Krishnan A, Ndungo E, Sandesara RG, Carette JE, Kuehne AI, Ruthel G, Pfeffer SR, Dye JM, Whelan SP, Brummelkamp TR, Chandran K. Ebola virus entry requires the host-programmed recognition of an intracellular receptor. EMBO J, 2012, 31: 1947–1960
White JM, Schornberg KL. A new player in the puzzle of filovirus entry. Nat Rev Microbiol, 2012, 10: 317–322
Chan SY, Speck RF, Ma MC, Goldsmith MA. Distinct mechanisms of entry by envelope glycoproteins of Marburg and Ebola (Zaire) viruses. J Virol, 2000, 74: 4933–4937
Ito H, Watanabe S, Sanchez A, Whitt MA, Kawaoka Y. Mutational analysis of the putative fusion domain of Ebola virus glycoprotein. J Virol, 1999, 73: 8907–8912
Lee JE, Fusco ML, Hessell AJ, Oswald WB, Burton DR, Saphire EO. Structure of the Ebola virus glycoprotein bound to an antibody from a human survivor. Nature, 2008, 454: 177–182
Feldmann H, Geisbert TW. Ebola haemorrhagic fever. Lancet, 2011, 377: 849–862
Ray RB, Basu A, Steele R, Beyene A, McHowat J, Meyer K, Ghosh AK, Ray R. Ebola virus glycoprotein-mediated anoikis of primary human cardiac microvascular endothelial cells. Virology, 2004, 321: 181–188
Yang ZY, Duckers HJ, Sullivan NJ, Sanchez A, Nabel EG, Nabel GJ. Identification of the Ebola virus glycoprotein as the main viral determinant of vascular cell cytotoxicity and injury. Nat Med, 2000, 6: 886–889
Sullivan N, Yang ZY, Nabel GJ. Ebola virus pathogenesis: implications for vaccines and therapies. J Virol, 2003, 77: 9733–9737
Sullivan NJ, Peterson M, Yang ZY, Kong WP, Duckers H, Nabel E, Nabel GJ. Ebola virus glycoprotein toxicity is mediated by a dynamin-dependent protein-trafficking pathway. J Virol, 2005, 79: 547–553
Zampieri CA, Fortin JF, Nolan GP, Nabel GJ. The ERK mitogen-activated protein kinase pathway contributes to Ebola virus glycoprotein-induced cytotoxicity. J Virol, 2006, 81: 1230–1240
Wahl-Jensen VM, Afanasieva TA, Seebach J, Stroher U, Feldmann H, Schnittler HJ. Effects of Ebola virus glycoproteins on endothelial cell activation and barrier function. J Virol, 2005, 79: 10442–10450
Geisbert TW, Young HA, Jahrling PB, Davis KJ, Kagan E, Hensley LE. Mechanisms underlying coagulation abnormalities in Ebola hemorrhagic fever: overexpression of tissue factor in primate monocytes/macrophages is a key event. J Infect Dis, 2003, 188: 1618–1629
Ruf W. Emerging roles of tissue factor in viral hemorrhagic fever. Trends Immunol, 2004, 25: 461–464
Bray M, Geisbert TW. Ebola virus: the role of macrophages and dendritic cells in the pathogenesis of Ebola hemorrhagic fever. Int J Biochem Cell Biol, 2005, 37: 1560–1566
Sobarzo A, Ochayon DE, Lutwama JJ, Balinandi S, Guttman O, Marks RS, Kuehne AI, Dye JM, Yavelsky V, Lewis EC, Lobel L. Persistent immune responses after Ebola virus infection. N Engl J Med, 2013, 369: 492–493
Boehm M, Slack FJ. microRNA control of lifespan and metabolism. Cell Cycle, 2006, 5: 837–840
Carleton M, Cleary MA, Linsley PS. microRNAs and cell cycle regulation. Cell Cycle, 2007, 6: 2127–2132
Scaria V, Hariharan M, Maiti S, Pillai B, Brahmachari SK. Host-virus interaction: a new role for microRNAs. Retrovirology, 2006, 3: 68
Lecellier CH, Dunoyer P, Arar K, Lehmann-Che J, Eyquem S, Himber C, Saib A, Voinnet O. A cellular microRNA mediates antiviral defense in human cells. Science, 2005, 308: 557–560
Motsch N, Pfuhl T, Mrazek J, Barth S, Grasser FA. Epstein-Barr virus-encoded latent membrane protein 1 (LMP1) induces the expression of the cellular microRNA miR-146a. RNA Biol, 2007, 4: 131–137
Song L, Liu H, Gao S, Jiang W, Huang W. Cellular microRNAs inhibit replication of the H1N1 influenza A virus in infected cells. J Virol, 2010, 84: 8849–8860
Trapnell C, Roberts A, Goff L, Pertea G, Kim D, Kelley DR, Pimentel H, Salzberg SL, Rinn JL, Pachter L. Differential gene and transcript expression analysis of RNA-seq experiments with Tophat and Cufflinks. Nat Prot, 2012, 7: 562–578
An J, Lai J, Lehman ML, Nelson CC. 2-miRDeep*: an integrated application tool for miRNA identification from RNA sequencing data. Nucleic Acids Res, 2013, 41: 727–737
Richardson JS, Yao MK, Tran KN, Croyle MA, Strong JE, Feldmann H, Kobinger GP. Enhanced protection against Ebola virus mediated by an improved adenovirus-based vaccine. PLoS One, 2009, 4: e5308
Jia YY, Gao P, Chen HZ, Wan YZ, Zhang R, Zhang ZQ, Yang RF, Wang X, Xu J, Liu DP. SIRT1 suppresses PMA and ionomycin-induced ICAM-1 expression in endothelial cells. Sci China Life Sci, 2013, 56: 19–25
Chan SY, Ma MC, Goldsmith MA. Differential induction of cellular detachment by envelope glycoproteins of Marburg and Ebola (Zaire) viruses. J Gen Virol, 2000, 81: 2155–2159
Provencal M, Michaud M, Beaulieu E, Ratel D, Rivard GE, Gingras D, Beliveau R. Tissue factor pathway inhibitor (TFPI) interferes with endothelial cell migration by inhibition of both the Erk pathway and focal adhesion proteins. Thromb Haemost, 2008, 99: 576–585
Chen J, Zhang B, Pan C, Ren L, Chen Y. Effects of monocyte chemotactic protein-3 on ICAM-1, VCAM-1, TF, and TFPI expression and apoptosis in human umbilical vein endothelial cells (in Chinese). Nan Fang Yi Ke Da Xue Xue Bao, 2013, 33: 86–92
Belkin AM, Smalheiser NR. Localization of cranin (dystroglycan) at sites of cell-matrix and cell-cell contact: recruitment to focal adhesions is dependent upon extracellular ligands. Cell Adhesion Commun, 1996, 4: 281–296
Park D, Shim E, Kim Y, Kim YM, Lee H, Choe J, Kang D, Lee YS, Jeoung D. C-FLIP promotes the motility of cancer cells by activating FAK and ERK, and increasing MMP-9 expression. Mol Cells, 2008, 25: 184–195
Winckers K, ten Cate H, Hackeng TM. The role of tissue factor pathway inhibitor in atherosclerosis and arterial thrombosis. Blood Rev, 2013, 27: 119–132
He MX, He YW. CFLAR/c-FLIPL: a star in the autophagy, apoptosis and necroptosis alliance. Autophagy, 2013, 9: 791–793
Karolchik D, Barber GP, Casper J, Clawson H, Cline MS, Diekhans M, Dreszer TR, Fujita PA, Guruvadoo L, Haeussler M, Harte RA, Heitner S, Hinrichs AS, Learned K, Lee BT, Li CH, Raney BJ, Rhead B, Rosenbloom KR, Sloan CA, Speir ML, Zweig AS, Haussler D, Kuhn RM, Kent WJ. The UCSC Genome Browser database: 2014 update. Nucleic Acids Res, 2014, 42: D764–770
Alazard-Dany N, Volchkova V, Reynard O, Carbonnelle C, Dolnik O, Ottmann M, Khromykh A, Volchkov VE. Ebola virus glycoprotein GP is not cytotoxic when expressed constitutively at a moderate level. J Gen Virol, 2006, 87: 1247–1257
Herquel B, Ouararhni K, Khetchoumian K, Ignat M, Teletin M, Mark M, Bechade G, van Dorsselaer A, Sanglier-Cianferani S, Hamiche A, Cammas F, Davidson I, Losson R. Transcription cofactors TRIM24, TRIM28, and TRIM33 associate to form regulatory complexes that suppress murine hepatocellular carcinoma. Proc Natl Acad Sci USA, 2011, 108: 8212–8217
Adams M. Tissue factor pathway inhibitor: new insights into an old inhibitor. Semin Thromb Hemost, 2012, 38: 129–134
Geisbert TW, Hensley LE, Jahrling PB, Larsen T, Geisbert JB, Paragas J, Young HA, Fredeking TM, Rote WE, Vlasuk GP. Treatment of Ebola virus infection with a recombinant inhibitor of factor VIIa/tissue factor: a study in rhesus monkeys. Lancet, 2003, 362: 1953–1958
Ledgerwood JE, Costner P, Desai N, Holman L, Enama ME, Yamshchikov G, Mulangu S, Hu Z, Andrews CA, Sheets RA, Koup RA, Roederer M, Bailer R, Mascola JR, Pau MG, Sullivan NJ, Goudsmit J, Nabel GJ, Graham BS. A replication defective recombinant Ad5 vaccine expressing Ebola virus GP is safe and immunogenic in healthy adults. Vaccine, 2010, 29: 304–313
Author information
Authors and Affiliations
Corresponding authors
Additional information
Contributed equally to this work
This article is published with open access at link.springer.com
Electronic supplementary material
Rights and permissions
Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0), which permits use, duplication, adaptation, distribution, and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
About this article
Cite this article
Sheng, M., Zhong, Y., Chen, Y. et al. Hsa-miR-1246, hsa-miR-320a and hsa-miR-196b-5p inhibitors can reduce the cytotoxicity of Ebola virus glycoprotein in vitro. Sci. China Life Sci. 57, 959–972 (2014). https://doi.org/10.1007/s11427-014-4742-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11427-014-4742-y