Abstract
Background
Ebola virus (EBOV) causes a serious hemorrhagic disease in humans, with a mortality rate of up to 80%. Despite significant achievements in the past decades elucidating the pathogenesis of EBOV, there is still much to be elucidated about the cell type-specific host response and their functional roles during infection.
Objective
This study aimed to gain insight into cell type-specific host responses to EBOV infection.
Methods
Real-time RT-qPCR analysis was used to identify host transcriptional changes in epithelial Caco-2 cells and endothelial HUVECs by EBOV infection.
Results
EBOV efficiently infected to both Caco-2 cells and HUVECs, depending on the time of infection. However, changes in the transcriptional levels of several host cellular genes following viral infection showed significant differences between Caco-2 cells and HUVECs. EBOV infection increases the transcription of TGF-β1, a key factor in epithelium-to-mesenchyme transition (EMT), only in HUVECs, but not in Caco-2 cells. This upregulation in turn induces the transcription of other EMT signaling molecules such as snail, slug and MMP9, ultimately leading to endothelial-to-mesenchymal transition (EndMT). Furthermore, this EndMT process appears to be associated with increased transcription of stem-cell markers such as Klf4, Sox2 and Oct4. However, most of these transcriptional changes due to EBOV infection did not occur in Caco-2 cells, suggesting that EMT or EndMT by EBOV infection is cell type-specific.
Conclusion
We propose that EBOV infection induces the expression of TGF-β1-mediated signals in endothelial HUVECs, resulting in EndMT. This could provide broader information to elucidate the pathogenesis of Ebola virus disease.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ansari AA (2014) Clinical features and pathobiology of Ebolavirus infection. J Autoimmun 55:1–9. https://doi.org/10.1016/j.jaut.2014.09.001
Bai X, Li YY, Zhang HY, Wang F, He HL, Yao JC, Liu L, Li SS (2017) Role of matrix metalloproteinase-9 in transforming growth factor-β1-induced epithelial-mesenchymal transition in esophageal squamous cell carcinoma. Onco Targets Ther 10:2837–2847. https://doi.org/10.2147/ott.s134813
Balda MS, Fallon MB, Van Itallie CM, Anderson JM (1992) Structure, regulation, and pathophysiology of tight junctions in the gastrointestinal tract. Yale J Biol Med 65:725–735
Batlle E, Sancho E, Francí C, Domínguez D, Monfar M, Baulida J, García De Herreros A (2000) The transcription factor snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol 2:84–89. https://doi.org/10.1038/35000034
Blow JA, Dohm DJ, Negley DL, Mores CN (2004) Virus inactivation by nucleic acid extraction reagents. J Virol Methods 119:195–198. https://doi.org/10.1016/j.jviromet.2004.03.015
Bose SK, Meyer K, Di Bisceglie AM, Ray RB, Ray R (2012) Hepatitis C virus induces epithelial-mesenchymal transition in primary human hepatocytes. J Virol 86:13621–13628. https://doi.org/10.1128/jvi.02016-12
Choi J, Park SY, Joo CK (2007) Transforming growth factor-β1 represses E-cadherin production via slug expression in lens epithelial cells. Invest Ophthalmol vis Sci 48:2708–2718. https://doi.org/10.1167/iovs.06-0639
De Craene B, Berx G (2013) Regulatory networks defining EMT during cancer initiation and progression. Nat Rev Cancer 13:97–110. https://doi.org/10.1038/nrc3447
Dianzani U, Malavasi F (1995) Lymphocyte adhesion to endothelium. Crit Rev Immunol 15:167–200. https://doi.org/10.1615/critrevimmunol.v15.i2.40
Feldmann H, Geisbert TW (2011) Ebola haemorrhagic fever. Lancet 377:849–862. https://doi.org/10.1016/s0140-6736(10)60667-8
Gaide Chevronnay HP, Selvais C, Emonard H, Galant C, Marbaix E, Henriet P (2012) Regulation of matrix metalloproteinases activity studied in human endometrium as a paradigm of cyclic tissue breakdown and regeneration. Biochim Biophys Acta 1824:146–156. https://doi.org/10.1016/j.bbapap.2011.09.003
Gao H, Teng C, Huang W, Peng J, Wang C (2015) SOX2 promotes the epithelial to mesenchymal transition of esophageal squamous cells by modulating slug expression through the activation of STAT3/HIF-α signaling. Int J Mol Sci 16:21643–21657. https://doi.org/10.3390/ijms160921643
Geisbert TW, Jahrling PB, Hanes MA, Zack PM (1992) Association of Ebola-related Reston virus particles and antigen with tissue lesions of monkeys imported to the United States. J Comp Pathol 106:137–152. https://doi.org/10.1016/0021-9975(92)90043-t
Gialeli C, Theocharis AD, Karamanos NK (2011) Roles of matrix metalloproteinases in cancer progression and their pharmacological targeting. FEBS J 278:16–27. https://doi.org/10.1111/j.1742-4658.2010.07919.x
Hajra KM, Chen DY, Fearon ER (2002) The SLUG zinc-finger protein represses E-cadherin in breast cancer. Cancer Res 62:1613–1618
Harcourt BH, Sanchez A, Offermann MK (1998) Ebola virus inhibits induction of genes by double-stranded RNA in endothelial cells. Virology 252:179–188. https://doi.org/10.1006/viro.1998.9446
Hargest V, Bub T, Neale G, Schultz-Cherry S (2022) Astrovirus-induced epithelial-mesenchymal transition via activated TGF-β increases viral replication. PLoS Pathog 18:e1009716. https://doi.org/10.1371/journal.ppat.1009716
Hemavathy K, Guru SC, Harris J, Chen JD, Ip YT (2000) Human Slug is a repressor that localizes to sites of active transcription. Mol Cell Biol 20:5087–5095. https://doi.org/10.1128/mcb.20.14.5087-5095.2000
Hochedlinger K, Yamada Y, Beard C, Jaenisch R (2005) Ectopic expression of Oct-4 blocks progenitor-cell differentiation and causes dysplasia in epithelial tissues. Cell 121:465–477. https://doi.org/10.1016/j.cell.2005.02.018
Jiang WG (1996) E-cadherin and its associated protein catenins, cancer invasion and metastasis. Br J Surg 83:437–446. https://doi.org/10.1002/bjs.1800830404
Jordà M, Olmeda D, Vinyals A, Valero E, Cubillo E, Llorens A, Cano A, Fabra A (2005) Upregulation of MMP-9 in MDCK epithelial cell line in response to expression of the Snail transcription factor. J Cell Sci 118:3371–3385. https://doi.org/10.1242/jcs.02465
Joseph MJ, Dangi-Garimella S, Shields MA, Diamond ME, Sun L, Koblinski JE, Munshi HG (2009) Slug is a downstream mediator of transforming growth factor-beta1-induced matrix metalloproteinase-9 expression and invasion of oral cancer cells. J Cell Biochem 108:726–736. https://doi.org/10.1002/jcb.22309
Kindrachuk J, Wahl-Jensen V, Safronetz D, Trost B, Hoenen T, Arsenault R, Feldmann F, Traynor D, Postnikova E, Kusalik A, Napper S, Blaney JE, Feldmann H, Jahrling PB (2014) Ebola virus modulates transforming growth factor β signaling and cellular markers of mesenchyme-like transition in hepatocytes. J Virol 88:9877–9892. https://doi.org/10.1128/jvi.01410-14
Kokudo T, Suzuki Y, Yoshimatsu Y, Yamazaki T, Watabe T, Miyazono K (2008) Snail is required for TGFβ-induced endothelial-mesenchymal transition of embryonic stem cell-derived endothelial cells. J Cell Sci 121:3317–3324. https://doi.org/10.1242/jcs.028282
Lebrin F, Goumans MJ, Jonker L, Carvalho RL, Valdimarsdottir G, Thorikay M, Mummery C, Arthur HM, ten Dijke P (2004) Endoglin promotes endothelial cell proliferation and TGF-β/ALK1 signal transduction. EMBO J 23:4018–4028. https://doi.org/10.1038/sj.emboj.7600386
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Martines RB, Ng DL, Greer PW, Rollin PE, Zaki SR (2015) Tissue and cellular tropism, pathology and pathogenesis of Ebola and Marburg viruses. J Pathol 235:153–174. https://doi.org/10.1002/path.4456
Medici D, Kalluri R (2012) Endothelial-mesenchymal transition and its contribution to the emergence of stem cell phenotype. Semin Cancer Biol 22:379–384. https://doi.org/10.1016/j.semcancer.2012.04.004
Medici D, Shore EM, Lounev VY, Kaplan FS, Kalluri R, Olsen BR (2010) Conversion of vascular endothelial cells into multipotent stem-like cells. Nat Med 16:1400–1406. https://doi.org/10.1038/nm.2252
Medici D, Potenta S, Kalluri R (2011) Transforming growth factor-β2 promotes Snail-mediated endothelial-mesenchymal transition through convergence of Smad-dependent and Smad-independent signalling. Biochem J 437:515–520. https://doi.org/10.1042/bj20101500
Michelson S, Alcami J, Kim SJ, Danielpour D, Bachelerie F, Picard L, Bessia C, Paya C, Virelizier JL (1994) Human cytomegalovirus infection induces transcription and secretion of transforming growth factor β1. J Virol 68:5730–5737. https://doi.org/10.1128/jvi.68.9.5730-5737.1994
Miettinen PJ, Ebner R, Lopez AR, Derynck R (1994) TGF-β induced transdifferentiation of mammary epithelial cells to mesenchymal cells: involvement of type I receptors. J Cell Biol 127:2021–2036. https://doi.org/10.1083/jcb.127.6.2021
Moonen JR, Krenning G, Brinker MG, Koerts JA, van Luyn MJ, Harmsen MC (2010) Endothelial progenitor cells give rise to pro-angiogenic smooth muscle-like progeny. Cardiovasc Res 86:506–515. https://doi.org/10.1093/cvr/cvq012
Munshi HG, Stack MS (2006) Reciprocal interactions between adhesion receptor signaling and MMP regulation. Cancer Metast Rev 25:45–56. https://doi.org/10.1007/s10555-006-7888-7
Nie Y, Cui D, Pan Z, Deng J, Huang Q, Wu K (2008) HSV-1 infection suppresses TGF-β1 and SMAD3 expression in human corneal epithelial cells. Mol vis 14:1631–1638
Park GB, Kim D, Kim YS, Kim S, Lee HK, Yang JW, Hur DY (2014) The Epstein-Barr virus causes epithelial-mesenchymal transition in human corneal epithelial cells via Syk/src and Akt/Erk signaling pathways. Invest Ophthalmol vis Sci 55:1770–1779. https://doi.org/10.1167/iovs.13-12988
Peinado H, Quintanilla M, Cano A (2003) Transforming growth factor β-1 induces snail transcription factor in epithelial cell lines: mechanisms for epithelial mesenchymal transitions. J Biol Chem 278:21113–21123. https://doi.org/10.1074/jbc.m211304200
Speranza E, Connor JH (2017) Host transcriptional response to ebola virus infection. Vaccines (basel) 5:30. https://doi.org/10.3390/vaccines5030030
Ströher U, West E, Bugany H, Klenk HD, Schnittler HJ, Feldmann H (2001) Infection and activation of monocytes by Marburg and Ebola viruses. J Virol 75:11025–11033. https://doi.org/10.1128/jvi.75.22.11025-11033.2001
Sun L, Diamond ME, Ottaviano AJ, Joseph MJ, Ananthanarayan V, Munshi HG (2008) Transforming growth factor-β1 promotes matrix metalloproteinase-9-mediated oral cancer invasion through snail expression. Mol Cancer Res 6:10–20. https://doi.org/10.1158/1541-7786.mcr-07-0208
Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126:663–676. https://doi.org/10.1016/j.cell.2006.07.024
Tamhankar M, Gerhardt DM, Bennett RS, Murphy N, Jahrling PB, Patterson JL (2018) Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells. Virol J 15:135. https://doi.org/10.1186/s12985-018-1045-0
Taylor MA, Parvani JG, Schiemann WP (2010) The pathophysiology of epithelial-mesenchymal transition induced by transforming growth factor-β in normal and malignant mammary epithelial cells. J Mammary Gland Biol Neoplasia 15:169–190. https://doi.org/10.1007/s10911-010-9181-1
Thiery JP, Acloque H, Huang RY, Nieto MA (2009) Epithelial-mesenchymal transitions in development and disease. Cell 139:871–890. https://doi.org/10.1016/j.cell.2009.11.007
Tiwari N, Meyer-Schaller N, Arnold P, Antoniadis H, Pachkov M, van Nimwegen E, Christofori G (2013) Klf4 is a transcriptional regulator of genes critical for EMT, including Jnk1 (Mapk8). PLoS ONE 8:e57329. https://doi.org/10.1371/journal.pone.0057329
van Meeteren LA, ten Dijke P (2012) Regulation of endothelial cell plasticity by TGF-β. Cell Tissue Res 347:177–186. https://doi.org/10.1007/s00441-011-1222-6
Wahl-Jensen V, Kurz S, Feldmann F, Buehler LK, Kindrachuk J, DeFilippis V, da Silva CJ, Früh K, Kuhn JH, Burton DR, Feldmann H (2011) Ebola virion attachment and entry into human macrophages profoundly effects early cellular gene expression. PLoS Negl Trop Dis 5:e1359. https://doi.org/10.1371/journal.pntd.0001359
Wesseling M, Sakkers TR, de Jager SCA, Pasterkamp G, Goumans MJ (2018) The morphological and molecular mechanisms of epithelial/endothelial-to-mesenchymal transition and its involvement in atherosclerosis. Vascul Pharmacol 106:1–8. https://doi.org/10.1016/j.vph.2018.02.006
Yao J, Guihard PJ, Blazquez-Medela AM, Guo Y, Moon JH, Jumabay M, Boström KI, Yao Y (2015) Serine protease activation essential for endothelial-mesenchymal transition in vascular calcification. Circ Res 117:758–769. https://doi.org/10.1161/circresaha.115.306751
Yori JL, Johnson E, Zhou G, Jain MK, Keri RA (2010) Kruppel-like factor 4 inhibits epithelial-to-mesenchymal transition through regulation of E-cadherin gene expression. J Biol Chem 285:16854–16863. https://doi.org/10.1074/jbc.m110.114546
Yu F, Li J, Chen H, Fu J, Ray S, Huang S, Zheng H, Ai W (2011) Kruppel-like factor 4 (KLF4) is required for maintenance of breast cancer stem cells and for cell migration and invasion. Oncogene 30:2161–2172. https://doi.org/10.1038/onc.2010.591
Zeisberg EM, Tarnavski O, Zeisberg M, Dorfman AL, McMullen JR, Gustafsson E, Chandraker A, Yuan X, Pu WT, Roberts AB, Neilson EG, Sayegh MH, Izumo S, Kalluri R (2007) Endothelial-to-mesenchymal transition contributes to cardiac fibrosis. Nat Med 13:952–961. https://doi.org/10.1038/nm1613
Zhang Y, Li C, Huang Y, Zhao S, Xu Y, Chen Y, Jiang F, Tao L, Shen X (2020) EOFAZ inhibits endothelial-to-mesenchymal transition through downregulation of KLF4. Int J Mol Med 46:300–310. https://doi.org/10.3892/ijmm.2020.4572
Zhang Y, Zhao S, Tu M, He L, Xu Y, Gan S, Shen X (2022) Inhibitory effect of essential oil from Fructus of Alpinia zerumbet on endothelial-to-mesenchymal transformation induced by TGF-β1 and downregulation of KLF4. J Cardiovasc Pharmacol 80:82–94. https://doi.org/10.1097/fjc.0000000000001283
Zhao Y, Qiao X, Wang L, Tan TK, Zhao H, Zhang Y, Zhang J, Rao P, Cao Q, Wang Y, Wang Y, Wang YM, Lee VW, Alexander SI, Harris DC, Zheng G (2016) Matrix metalloproteinase 9 induces endothelial-mesenchymal transition via Notch activation in human kidney glomerular endothelial cells. BMC Cell Biol 17:21. https://doi.org/10.1186/s12860-016-0101-0
Zhou Q, Fan D, Huang K, Chen X, Chen Y, Mai Q (2018) Activation of KLF4 expression by small activating RNA promotes migration and invasion in colorectal epithelial cells. Cell Biol Int 42:495–503. https://doi.org/10.1002/cbin.10926
Funding
No funding was received.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that there is no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Ro, YT., Patterson, J.L. Transcriptional induction of TGF-β1 and endothelial-to-mesenchymal transition cell markers in human umbilical vein endothelial cells by Ebola virus infection. Genes Genom 44, 1499–1507 (2022). https://doi.org/10.1007/s13258-022-01333-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13258-022-01333-x