Cytoprotective Effect of Recombinant Human Erythropoietin Produced in Transgenic Tobacco Plants

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dc.contributor.authorKittur, Farooqahmed S.en
dc.contributor.authorBah, Mamudouen
dc.contributor.authorArcher-Hartmann, Stephanieen
dc.contributor.authorHung, Chiu-Yuehen
dc.contributor.authorAzadi, Parastooen
dc.contributor.authorIshihara, Mayumien
dc.contributor.authorSane, David C.en
dc.contributor.authorXie, Jiahuaen
dc.date.accessioned2018-10-17T14:56:40Zen
dc.date.available2018-10-17T14:56:40Zen
dc.date.issued2013-10-04en
dc.description.abstractAsialo-erythropoietin, a desialylated form of human erythropoietin (EPO) lacking hematopoietic activity, is receiving increased attention because of its broader protective effects in preclinical models of tissue injury. However, attempts to translate its protective effects into clinical practice is hampered by unavailability of suitable expression system and its costly and limit production from expensive mammalian cell-made EPO (rhuEPOM) by enzymatic desialylation. In the current study, we took advantage of a plant-based expression system lacking sialylating capacity but possessing an ability to synthesize complex N-glycans to produce cytoprotective recombinant human asialo-rhuEPO. Transgenic tobacco plants expressing asialo-rhuEPO were generated by stably co-expressing human EPO and β1,4-galactosyltransferase (GalT) genes under the control of double CaMV 35S and glyceraldehyde-3-phosphate gene (GapC) promoters, respectively. Plant-produced asialo-rhuEPO (asialo-rhuEPOP) was purified by immunoaffinity chromatography. Detailed N-glycan analysis using NSI-FTMS and MS/MS revealed that asialo-rhuEPOP bears paucimannosidic, high mannose-type and complex N-glycans. In vitro cytoprotection assays showed that the asialo-rhuEPOP (20 U/ml) provides 2-fold better cytoprotection (44%) to neuronal-like mouse neuroblastoma cells from staurosporine-induced cell death than rhuEPOM (21%). The cytoprotective effect of the asialo-rhuEPOP was found to be mediated by receptor-initiated phosphorylation of Janus kinase 2 (JAK2) and suppression of caspase 3 activation. Altogether, these findings demonstrate that plants are a suitable host for producing cytoprotective rhuEPO derivative. In addition, the general advantages of plant-based expression system can be exploited to address the cost and scalability issues related to its production.en
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1371/journal.pone.0076468en
dc.identifier.eissn1932-6203en
dc.identifier.issue10en
dc.identifier.othere76468en
dc.identifier.pmid24124563en
dc.identifier.urihttp://hdl.handle.net/10919/85393en
dc.identifier.volume8en
dc.language.isoenen
dc.publisherPLOSen
dc.rightsCreative Commons Attribution 4.0en
dc.rights.urihttps://creativecommons.org/licenses/by/4.0en
dc.titleCytoprotective Effect of Recombinant Human Erythropoietin Produced in Transgenic Tobacco Plantsen
dc.title.serialPLOS ONEen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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