Retinoic Acid-Loaded Nanoparticles Promote Neurovascular Protection in Stroke.

HomeStrokeVol. 54, No. 4Retinoic Acid–Loaded Nanoparticles Promote Neurovascular Protection in Stroke Free AccessResearch ArticlePDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissionsDownload Articles + Supplements ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toSupplemental MaterialFree AccessResearch ArticlePDF/EPUBRetinoic Acid–Loaded Nanoparticles Promote Neurovascular Protection in Stroke Marta Machado-Pereira, Alba Grayston, Miguel Garcia-Gabilondo, Vitor Francisco, Ana Cristóvão, João Marto, Helena Vieira, Miguel Viana-Baptista, Lino Ferreira, Liliana Bernardino, Anna Rosell and Raquel Ferreira Marta Machado-PereiraMarta Machado-Pereira https://orcid.org/0000-0002-8986-8679 Health Sciences Research Centre (M.M.-P., A.C., L.B., R.F.), University of Beira Interior, Portugal. Search for more papers by this author , Alba GraystonAlba Grayston https://orcid.org/0000-0002-1466-0099 Health Sciences Research Centre (M.M.-P., A.C., L.B., R.F.), University of Beira Interior, Portugal. Neurovascular Research Laboratory, Vall d’Hebron Institut de Recerca, Universitat Autónoma de Barcelona, Spain (A.G., M.G.-G., A.R.). Search for more papers by this author , Miguel Garcia-GabilondoMiguel Garcia-Gabilondo https://orcid.org/0000-0002-2015-2621 Neurovascular Research Laboratory, Vall d’Hebron Institut de Recerca, Universitat Autónoma de Barcelona, Spain (A.G., M.G.-G., A.R.). Search for more papers by this author , Vitor FranciscoVitor Francisco https://orcid.org/0000-0002-2704-9950 Center for Neuroscience and Cell Biology (V.F., L.F.), University of Coimbra, Portugal. Faculty of Medicine (V.F., L.F.), University of Coimbra, Portugal. Search for more papers by this author , Ana CristóvãoAna Cristóvão https://orcid.org/0000-0001-5806-4478 NeuroSoV, UBImedical (A.C.), University of Beira Interior, Portugal. Search for more papers by this author , João MartoJoão Marto https://orcid.org/0000-0003-2277-5950 Department of Neurology, Hospital Egas Moniz, Centro Hospitalar de Lisboa Ocidental, Portugal (J.M., M.V.-B.). NMS Research, Nova Medical School Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Portugal (J.M., H.V., M.V.-B., R.F.). Search for more papers by this author , Helena VieiraHelena Vieira https://orcid.org/0000-0001-9415-3742 NMS Research, Nova Medical School Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Portugal (J.M., H.V., M.V.-B., R.F.). Applied Molecular Biosciences Unit (H.V.), NOVA School of Science and Technology, Universidade NOVA de Lisboa, Portugal. Associate Laboratory i4HB – Institute for Health and Bioeconomy, Department of Chemistry (H.V.), NOVA School of Science and Technology, Universidade NOVA de Lisboa, Portugal. Search for more papers by this author , Miguel Viana-BaptistaMiguel Viana-Baptista https://orcid.org/0000-0001-6166-2073 Department of Neurology, Hospital Egas Moniz, Centro Hospitalar de Lisboa Ocidental, Portugal (J.M., M.V.-B.). NMS Research, Nova Medical School Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Portugal (J.M., H.V., M.V.-B., R.F.). Search for more papers by this author , Lino FerreiraLino Ferreira https://orcid.org/0000-0001-8985-9302 Center for Neuroscience and Cell Biology (V.F., L.F.), University of Coimbra, Portugal. Faculty of Medicine (V.F., L.F.), University of Coimbra, Portugal. Search for more papers by this author , Liliana BernardinoLiliana Bernardino Liliana Bernardino, PhD, Health Sciences Research Centre, University of Beira Interior, Portugal, Email E-mail Address: [email protected] https://orcid.org/0000-0003-3395-5973 Health Sciences Research Centre (M.M.-P., A.C., L.B., R.F.), University of Beira Interior, Portugal. Search for more papers by this author , Anna RosellAnna Rosell Neurovascular Research Laboratory, Vall d’Hebron Institut de Recerca, Universitat Autónoma de Barcelona, Spain (A.G., M.G.-G., A.R.). Search for more papers by this author and Raquel FerreiraRaquel Ferreira Correspondence to: Raquel Ferreira, PhD, NOVA Medical School Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Campo dos Mártires da Pátria 130, 1169-056 Portugal, Email E-mail Address: [email protected] https://orcid.org/0000-0003-1014-0499 Health Sciences Research Centre (M.M.-P., A.C., L.B., R.F.), University of Beira Interior, Portugal. NMS Research, Nova Medical School Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Portugal (J.M., H.V., M.V.-B., R.F.). Now with Technophage, Lisbon, Portugal (R.F.). Search for more papers by this author Originally published13 Mar 2023https://doi.org/10.1161/STROKEAHA.122.041839Stroke. 2023;54:e149–e151Other version(s) of this articleYou are viewing the most recent version of this article. Previous versions: March 13, 2023: Ahead of Print The role of Ago2 (argonaute-2) in stroke is insufficiently described. Loss of this increases endothelial cell death and impairs tubule formation.1 Conversely, the abnormal human brain vasculature overexpresses Ago2, and we have highlighted its clinical potential as a systemic microRNA carrier; signaling pathways remain unexplored in stroke.2 The actions of retinoic acid (RA) are complex to manage (poor solubility, photosensitivity, narrow therapeutic window), but its encapsulation in trackable polymeric nanoparticles (RA polymeric nanoparticles [RA-NP]) has enhanced its efficacy.3 In vitro, 3 µg/mL RA-NP (not the free molecule [0.12 µM]), recuperated Ago2 after oxygen and glucose deprivation, followed by a recovery period intended to mimic reperfusion (OGD/R; Figure [A]). Normalizing intracellular Ago2 in an inflammatory context recuperates the neurovascular unit, via the NO pathway, which also intersects with RA signaling.3,4 After OGD/R, RA-NP normalized the phosphorylation of Akt and overexpression of endothelial nitric oxide synthase (eNOS; Figure [B]) and consequent NO overproduction (Figure [C]). By silencing Ago2 or by inhibiting its activity, NO production was also enhanced, producing the first evidence that RA-NP can operate via Ago2 and the regulation of the Akt (protein kinase B)/eNOS/NO pathway. In vivo,5 a single intravenous injection of RA-NP (20 µg/g), equivalent to 0.246 µg/g free RA, reached the brain without systemic toxicity and reduced infarct volume, three days poststroke (Figure [D]). In this period, RA-NP normalized microglial anti-ionized calcium binding adaptor molecule-1 and astrocytic GFAP (anti-glial fibrillary acidic protein) levels (Figure [E]), when inflammation is more exacerbated. Seven days poststroke, treated mice recovered grip strength (Figure [D]), and in parallel, RA-NP increased vessel density (Figure [F]), suggesting a promising proangiogenic effect in the peri-infarct region. Therefore, RA-NP could be envisioned as a potential therapeutic systemic agent aimed at treating neurovascular and/or neuroinflammatory conditions.Download figureDownload PowerPointFigure. Retinoic acid polymeric nanoparticles (RA-NP) modulate markers relevant for the recovery of the ischemic brain. RA-NP recuperated Ago2 (argonaute-2; A), pAkt (phosphorylated- Akt), and eNOS (B) in primary brain endothelial cells after oxygen and glucose deprivation followed by a recovery period (OGD/R), resulting in normalized NO production (C). Using a model of transient middle cerebral artery occlusion, RA-NP reduced infarct volume and increased grip strength (D). RA-NP normalized microglial and astrocytic markers (anti-ionized calcium binding adaptor molecule-1 [Iba-1] and GFAP [anti-glial fibrillary acidic protein]) 3 days postinjection (E), and increased vessel density compromised by stroke 7 days postinjection (F). CT indicates contralateral; IP, ipsilateral; and siAgo2, small interfering ribonucleic acid for argonaute-2.MethodsExpanded Methods are detailed in the in Supplemental Material.4,5 All experiments were performed in accordance with the National Institutes of Health and European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes (European Union directive number 2010/63/EU) for the care and use of laboratory animals, the Ethics Committee for Animal Experimentation of the Vall d’Hebron Research Institute, according to Spanish legislation.The data that support the findings of this study are available from the corresponding author upon reasonable request, according to the American Heart Association Journals’ implementation of the Transparency and Openness Promotion Guidelines.Article InformationSources of FundingSupported by Portuguese Platform of BioImaging, POCI-01-0145-FEDER-022122; FCT, UID/Multi/00709/2013, SFRH/BD/137440/2018, ISCIII, FI17/00073; PTDC/BTM- SAL/5174/2020, EXPL/BTM-ORG/1348/2021; INTER-REG Atlantic Area (EAPA_791/2018_ NEUROATLANTIC Project), INTER-REG V A España Portugal (POCTEP; 0624_2IQBIONEURO_6_E), European Regional Development Fund; SLT017/20/000197 from AGAUR; RICORS-Stroke Network from ISCIII (RD21/0006/0007); IF/00178/2015/CP1300/CT0001; PERIS-SLT017/20/000197 (Generalitat Catalunya).Supplemental MaterialSupplemental MethodsDisclosures None.FootnotesFor Sources of Funding and Disclosures, see page e150.Supplemental Material is available at https://www.ahajournals.org/doi/suppl/10.1161/STROKEAHA.122.041839.Correspondence to: Raquel Ferreira, PhD, NOVA Medical School Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Campo dos Mártires da Pátria 130, 1169-056 Portugal, Email raquel.ferreira@nms.unl.ptLiliana Bernardino, PhD, Health Sciences Research Centre, University of Beira Interior, Portugal, Email libernardino@fcsaude.ubi.ptReferences1. Asai T, Suzuki Y, Matsushita S, Yonezawa S, Yokota J, Katanasaka Y, Ishida T, Dewa T, Kiwada H, Nango M, et al. Disappearance of the angiogenic potential of endothelial cells caused by Argonaute2 knockdown.Biochem Biophys Res Commun. 2008; 368:243–248. doi: 10.1016/j.bbrc.2008.01.074CrossrefGoogle Scholar2. Ferreira R, Santos T, Amar A, Gong A, Chen TC, Tahara SM, Giannotta SL, Hofman FM. Argonaute-2 promotes miR-18a entry in human brain endothelial cells.J Am Heart Assoc. 2014; 3:e000968. doi: 10.1161/JAHA.114.000968LinkGoogle Scholar3. Ferreira R, Napoli J, Enver T, Bernardino L, Ferreira L. Advances and challenges in retinoid delivery systems in regenerative and therapeutic medicine.Nat Commun. 2020; 11:1–14. doi: 10.1038/s41467-020-18042-2CrossrefGoogle Scholar4. Machado-Pereira M, Saraiva C, Bernardino L, Cristóvão AC, Ferreira R. Argonaute- 2 protects the neurovascular unit from damage caused by systemic inflammation.J Neuroinflammation. 2022; 19:1–17. doi: 10.1186/s12974-021-02324-7CrossrefGoogle Scholar5. Morancho A, García-Bonilla L, Barceló V, Giralt D, Campos-Martorell M, Garcia S, Montaner J, Rosell A. A new method for focal transient cerebral ischaemia by distal compression of the middle cerebral artery.Neuropathol Appl Neurobiol. 2012; 38:617–627. doi: 10.1111/j.1365-2990.2012.01252.xCrossrefMedlineGoogle Scholar eLetters(0)eLetters should relate to an article recently published in the journal and are not a forum for providing unpublished data. Comments are reviewed for appropriate use of tone and language. Comments are not peer-reviewed. Acceptable comments are posted to the journal website only. Comments are not published in an issue and are not indexed in PubMed. Comments should be no longer than 500 words and will only be posted online. References are limited to 10. Authors of the article cited in the comment will be invited to reply, as appropriate.Comments and feedback on AHA/ASA Scientific Statements and Guidelines should be directed to the AHA/ASA Manuscript Oversight Committee via its Correspondence page.Sign In to Submit a Response to This Article Previous Back to top Next FiguresReferencesRelatedDetails April 2023Vol 54, Issue 4 Advertisement Article InformationMetrics © 2023 American Heart Association, Inc.https://doi.org/10.1161/STROKEAHA.122.041839PMID: 36912140 Originally publishedMarch 13, 2023 Keywordsnanoparticlesribonucleoproteininflammationendothelial cellstrokePDF download Advertisement SubjectsCell Signaling/Signal TransductionCerebrovascular Disease/StrokeIschemic Stroke