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siRNA-cell-penetrating peptides complexes as a combinatorial therapy against chronic myeloid leukemia using BV173 cell line as model

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dc.contributor.authorFreire, João Miguel
dc.contributor.authorFigueiredo, Inês Rego de
dc.contributor.authorValle, Javier
dc.contributor.authorVeiga, Ana Salomé
dc.contributor.authorAndreu, David
dc.contributor.authorEnguita, Francisco J.
dc.contributor.authorCastanho, Miguel A. R. B.
dc.date.accessioned2018-04-03T11:50:53Z
dc.date.available2018-04-03T11:50:53Z
dc.date.issued2017
dc.description© 2016 Elsevier B.V. All rights reserved.pt_PT
dc.description.abstractChronic myeloid leukemia (CML) is a myeloproliferative disorder caused by a single gene mutation, a reciprocal translocation that originates the Bcr-Abl gene with constitutive tyrosine kinase activity. As a monogenic disease, it is an optimum target for RNA silencing therapy. We developed a siRNA-based therapeutic approach in which the siRNA is delivered by pepM or pepR, two cell-penetrating peptides (CPPs) derived from the dengue virus capsid protein. These peptides have a dual role: siRNA delivery into cells and direct action as bioportides, i.e. intracellularly bioactive CPPs, targetting cancer-related signaling processes. Both pepM and pepR penetrate the positive Bcr-Abl+ Cell Line (BV173). Five in silico designed anti-Bcr-Abl siRNA were selected for in vitro analysis after thorough screening. The Bcr-Abl downregulation kinetics (48h to 168h) was followed by quantitative PCR. The bioportide action of the peptide vectors was evaluated by genome-wide microarray analysis and further validated by testing BV173 cell cycle and cell proliferation monitoring different genes involved in housekeeping/cell stress (RPL13A, HPRT1), cell proliferation (ki67), cell apoptosis (Caspase 3 and Caspase 9) and cell cycle steps (CDK2, CCDN2, CDKN1A). Assays with a commercial transfection agent were carried out for comparison purposes. Maximal Bcr-Abl gene knockdown was observed for one of the siRNA when delivered by pepM at 120h. Both pepM and pepR showed downregulation effects on proliferative CML-related signaling pathways having direct impact on BV173 cell cycle and proliferation, thus reinforcing the siRNA effect by acting as anticancer molecules. With this work we show the therapeutic potential of a CPP shuttle that combines intrinsic anticancer properties with the ability to deliver functional siRNA into CML cell models. By such combination, the pepM-siRNA conjugates lowered Bcr-Abl gene expression levels more extensively than conventional siRNA delivery technologies and perturbed leukemogenic cell homeostasis, hence revealing their potential as novel alternative scaffolds for CML therapy.pt_PT
dc.description.sponsorshipThis work was supported by Fundação para a Ciência e Tecnologia –Ministério da Educação e Ciência (FCT-MEC, Portugal) [PTDC/QUI-BIQ/112929/2009], by Gabinete de Apoio à Investigação Científica, Tecnológica e Inovação (GAPIC – grants n° 20110007 and 20120006), Fundação AstraZeneca Innovative Competition 2013 (iMed 5.0), the European Union [FP7-HEALTH-F3-2008-223414 (LEISHDRUG)], by the Spanish Ministry of Economy and Competitiveness (SAF2011-24899) and the Generalitat de Catalunya (2009 SGR 492). JMF acknowledges FCT-MEC for PhD fellowship SFRH/BD/70423/2010 and ASV for fellowship IF/00803/2012 under the FCT Investigator Programmept_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationJournal of Controlled Release 245 (2017) 127–136pt_PT
dc.identifier.doi10.1016/j.jconrel.2016.11.027pt_PT
dc.identifier.issn0168-3659
dc.identifier.urihttp://hdl.handle.net/10451/32559
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relationFP7-HEALTH-F3-2008-223414pt_PT
dc.relationIF/00803/2012pt_PT
dc.relationSTRUCTURAL PROTEINS OF ENVELOPED VIRUS: FROM BIOCHEMISTRY TO MOLECULAR MEDICINE. A FOCUS ON DENGUE HEMORRHAGIC FEVER VIRUS
dc.relation.publisherversionhttps://www.journals.elsevier.com/journal-of-controlled-release/pt_PT
dc.subjectBcr-Ablpt_PT
dc.subjectBioportidept_PT
dc.subjectCell-penetrating peptidept_PT
dc.subjectChronic myeloid leukemiapt_PT
dc.subjectDengue virus capsid proteinpt_PT
dc.subjectMicroarraypt_PT
dc.subjectsiRNApt_PT
dc.titlesiRNA-cell-penetrating peptides complexes as a combinatorial therapy against chronic myeloid leukemia using BV173 cell line as modelpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardNumberPTDC/QUI-BIQ/112929/2009
oaire.awardNumberSFRH/BD/70423/2010
oaire.awardTitleSTRUCTURAL PROTEINS OF ENVELOPED VIRUS: FROM BIOCHEMISTRY TO MOLECULAR MEDICINE. A FOCUS ON DENGUE HEMORRHAGIC FEVER VIRUS
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FQUI-BIQ%2F112929%2F2009/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/OE/SFRH%2FBD%2F70423%2F2010/PT
oaire.citation.endPage136pt_PT
oaire.citation.startPage127pt_PT
oaire.citation.titleJournal of Controlled Releasept_PT
oaire.citation.volume245pt_PT
oaire.fundingStream3599-PPCDT
oaire.fundingStreamOE
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT
relation.isProjectOfPublication954cb0bb-03f8-45f6-a48d-27c88a2f8690
relation.isProjectOfPublication740ee52c-b5d2-4811-9651-6283006aebee
relation.isProjectOfPublication.latestForDiscovery740ee52c-b5d2-4811-9651-6283006aebee

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