Results:

► Design, preparation and characte-rization of siRNA / shRNA-carryingnanoparticles ( siRNA/ shRNA-NPs)targeted to P-selectin expressed onvalvular endothelial cells surface (VEC)(patent application OSIM nr. A/00811,Constantinescu CA et al., Pharmaceutics,2019). These P-selectin targeted siRNA /shRNA nanocarriers bind to cultured EC andaortic valve of ApoE-deficient mice.

Ongoing experiments are designed tooptimize nanocarriers targeted to P-selectinor other molecules, such as VCAM-1 orcollagen IV to obtain maximal transfectionefficiency of siRNA/shRNA in VEC and VIC

exposed to high glucose conditions in thepresence or absence of osteogenic factors.Specific shRNA sequences for relevantmoleculesinvolved in EndMT and VICosteodifferentiation are envisaged for theevaluation of the in vitro and invivotherapeutic effects of targetednanocarriers.

PERSPECTIVES

● Design of novel nanoparticle-based drugdelivery systems to specifically target inflamedendothelium and monocytes/macrophages and tomodulate their affected functions;

● The development of new targetednanotherapies for aortic valve disease.

Ongoing experiments are focused on theinvestigation of the in vivo therapeutic effectsof endothelium-targeted polyphenols-loadedLN in appropriate pre-clinical animal models.

2. Development of nano-therapeutics for targeteddelivery of siRNA/shRNA toaortic valve to down-regulate genes that drivesaortic valve disease

Recent data suggest that in the early phaseof aortic valve disease (AVD), inflammationcan determine a subset of aortic valvularendothelial cells (VEC) to undergo endothelialto mesenchymal transformation (EndMT) and

also, can initiate the osteodifferentiation ofvalvular interstitial cells (VIC) that activelycontribute to aortic valve calcification.

We hypothesize that the blocking of the twokey features of aortic valve lesions, namelyEndMT and VIC’s osteodifferentiation usingnanocarriers of siRNA/shRNA todownregulate genes that drive EndMT (e.g.SMAD3, SNAI1) and VIC osteodifferentiation(e.g. RUNX2)may represent a therapeuticstrategy with an impact for early stages ofAVD.

The objective is to design effectivenanocarriers of siRNA/shRNA able to targetvalvular cells or extracellular matrix indiabetes-affected aortic valve and to blockEndMT and VIC to osteoblasts transformation.

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targeted delivery of siRnA/shRnA to valvular cells using nanocarriers.

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(A) Zetasizer nano ZS (Malvern Instruments, uK)used for (B) size and zeta potential measurements ofsiRnA/shRnA nanocarriers; (C) the accumulationof fluorescently labelled P-selectin targetedlipoplexes (c) and non-targeted lipoplexes coupledwith a scrambled peptide (b) to the aortic valve ofApoe-deficient mice. Control aortic valve from amouse injected with PbS (a); (D) Spectral unmixingshowing the autofluorescence of the tissue (green)and the specific accumulation of P-selectin targetedfluorescently-labelled lipoplexes (red) in the aorticvalve of Apoe-deficient mouse.

Medical and Pharmaceutical BioNanoTechnologies Laboratory

INTERNATIONAL

● Cardiovascular Tissue Engineering andRegenerative Medicine Lab, ClemsonUniversity, USA (Dan Simionescu)

● Biocompatibility and TissueRegeneration Lab, Clemson University, USA(Agneta Simionescu)

● Department of Clinical Sciences,Malmö, Lund University, Sweden (AlexandruSchiopu)

● Department of KardiovaskuläreMolekularbiologie, UniversitätsklinikumAachen, Germany (Christian Weber, RoryKoenen)

● Department of Pharmacy, Martin-LutherUniversity, Halle, Germany (Gerd Bendas)

● Department of Pharmacy, University ofBonn, Germany (Gerd Bendas, MartinSchlesinger)

● Faculty of Pharmacy, University ofIstanbul, Turkey (Erdal Cevher)

● Institute of Physiology, University ofZürich, Switzerland (Lubor Borsig, MarkoRoblek)

● University of Minho, Portugal (SandraCarvahlo)

● Montana University of Leoben, Austria(Robert Franz)

NATIONAL

● Institute of Cardiovascular Disease“Prof. Dr. C.C. Iliescu”,Bucharest (CarmenGinghina)

● Emergency Hospital of M.A.I. Prof. Dr.D. Gerota, Bucharest, Romania (MonicaCăpraru)

● University of Medicine and Pharmacy”Carol Davila”, Bucharest, România (DragoșVinereanu)

● Center of Surface Science andNanotechnology, University Politehnica ofBucharest (Marius Enăchescu)

● Centrul Clinic de Urgenţă deBoliCardiovasculare”Dr. Constantin Zamfir”(Ionel Droc)

● The Institute for Research andDevelopment of Textiles and Leather,Bucharest (Carmen Gaidau)

● Institute of Macromolecular Chemistry“Petru Poni, Iași (Mariana Pinteală)

● National Institute of Materials Physics(Ionuț Enculescu)

● Department of Natural and SyntheticPolymers, “Gh. Asachi” Technical Universityof Iași (Geta David)

● SC Optoelectronica 2000 SRL,Magurele (Teodor Necșoiu, MihaiȘerbănescu)

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COLLABORATIONof the department

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GRANTS AWARDED BYCOmpetition(1999-2019)

● 2018-2022:PCCf project, cod PN-III-P4-ID-PCCf-2016-0172 (INNATE-MI):“targeting innate immune mechanisms to improverisk stratification and to identify future therapeuticoptions in myocardial infarction”, projectcoordinator: Acad. Maya Simionescu - IBPC “N.Simionescu. Partners: University of Medicine,Pharmacy, Science and Technology of TârguMureş, “CAROL DAVILA” University ofMedicine and Pharmacy Bucharest.

● 2018-2022:PCCf project, cod PN-III-P4-ID-PCCf-2016-0050 (5D-nanoP),“Mimickingliving matter mechanisms by five-dimensionalchemistry approaches” partners:Institute ofMacro-molecular Chemistry “Petru Poni, Iași;IBPC “N. Simionescu”, Bucharest; Center forOrganic Chemistry “Costin D. Neniţescu”,Bucharest (partner responsible: Maya Simionescuand Manuela Călin).

● 2018-2020: PCCDI Complex Project nr.13 PCCDI/2018(INTERA) “Intelligenttherapies for non-communicable diseases basedon controlled release of pharma-cologicalcompounds from encapsulated engineered cellsand targeted bionanoparticles“ (complex projectcoordinator: Maya Simionescu), INTERA2component project: “Development of a 3Dplatform designed for pre-clinical drug testingcomposed of cells incorporated into three-dimensional bio-matrices”, (project INTERA2coordinator: Elena Butoi); INTERA3 componentproject:” Intelligent nanobioparticles designed tofunction as vectors for targeted delivery ofbioactive compounds in vascular inflammationtherapy” (project INTERA3 coordinator:Manuela Călin).

● 2014-2020: Competitiveness Opera-tional Programme Project, PriorityAxis1/Action 1.1.4 (THERAVALDIS):“targeted therapies for diabetes -relatedaortic valve disease”,Financing Contractno.115/13.09.2016/ MySMIS: 104362 -executive manager: Ileana Mânduțeanu.

● 2017-2019: ELI-RO/ PN-III-P5-Subprogramme 5.1, on-line measurement oflaser-driven proton beams effect on human cells.Project coordinator Adrian Enache- NationalInstitute of Materials Physics, (ICBP- NSimionescu, partner responsible: Elena Butoi).

● 2015-2018: SIINN ERA-NET projectfP7 scheme (NANO_SAfE_LEATHER):“the effect on human health of Ag/tio2 nM-treated leathers for footwear industry”, partners:The Institute for Research and Development ofTextiles and Leather, Romania; ICBP “NicolaeSimionescu”, România; University of Minho,Portugal; Montana University of Leoben,Austria; SC TARO COMMIMPEX LTD,România (partner responsible: Manuela Călin).

● 2015-2017: grant PN-II-RU-TE-2014-4-0965, “MicroRnA signature of vascular cellscross-talk relevant for the atherosclerotic plaquerupture in patients with type II diabetes” (projectcoordinator: Elena Butoi).

● 2015-2017: PNCDI II grant nr. PN-II-RU-TE-2014-4-1837 (funded by UEFISCDI):“endothelium-targeted nano-therapies desig-ned to silence receptor for advanced glycationproducts (RAGe) in atherosclerosis”, (projectcoordinator: Manuela Călin).

● 2011-2014: EuroNanoMed ERA-NETproject fP7 scheme (NANODIATER)“nanoparticles designed to target chemokine-related inflammatory processes in vasculardiseases and cancer metastasis andimplementation of a biosensor to diagnose thesedisorders”, partners: ICBP “Nicolae Simionescu”,Romania; Center of Surface Science andNanotechnology, University Polyethnic ofBucharest; University of Bonn, Germany; IstanbulUniversity, Turkey; University of Zürich,Switzerland;EPO Berlin GmbH , Germany; SCOptoelectronica 2000 SRL (project director: MayaSimionescu; Research & Developmentcoordinator: Manuela Călin).

● 2011-2014: Project PNII: IDEAS:”Molecules and mechanisms involved invascular inflammation dependent on cytokinesand chemokines as possible targets for new

Medical and Pharmaceutical BioNanoTechnologies Laboratory

nanoterapeutic strategies”, (project coordinator:Ileana Mânduțeanu).

● 2007-2010: Project PNCDI2- P4project: “Molecular links between chronicinflammation and accelerated athero-sclerosis:role of resistin and the newly identifiedchemokines:fractalkine and CXCL16:newavenues for targeted therapies”, (projectcoordinator: Ileana Mânduțeanu).

● 2006-2008: National Authority forScientific Research and Innovation (ANCS)grant nr.1423/2006: “Study of signalingpathways involved in hyperglycemia - inducedfractalkine expression and their targeting, a newapproach to the therapy of cardiovascularpathologies associated with diabetes” (projectcoordinator: Manuela Călin)

● 2004-2006: VIASAN PNCDI grant, nr.330/2004: “A new strategy to stabilize theatherosclerotic lesions in acute coronarysyndromes: suppression of activated macrophagesusing clodronate-loaded liposomes”(projectcoordinator: Manuela Călin)

● 2005-2008: fP6 International project,Specific Support Action (SERA), Memberin the management cometee, WP LeaderIleana Mânduțeanu.

● 2003-2005: grant VIASAN, MEC:“the chemokine modulation in differentvascular pathologies; their functional role”,(project coordinator: Elena Butoi).

● 2001-2003: VIASAN PNCDI grant,nr. 031/2001: “targeted delivery of drugs toactivated endothelium using “smart”liposomes: a strategy for cardiovasculardiseases therapy” (project coordinator:Manuela Călin).

● 2003-2004: Romanian Ministry ofEducation and Research grant:“Protective effects of aspirin in diabetesmellitus model, in vitro” (project coordinator:Elena Butoi).

● 2003-2004: Romanian Academygrant: “Study of the effect of superoxide

dismutase administered in liposomes on thereactivity of mesenteric arteries isolated fromdiabetic hamsters”(project coordinator:Manuela Călin).

● 2001-2002: Romanian Ministry ofEducation and Research grant: “the effectof the anti-inflammatory drugs on theactivated vascular endothelium” (projectcoordinator: Elena Butoi).

● 1999-2001: National Agency forScience, Technology and Innovation (ANSTI)grant nr.5243/1999: “Specific drug delivery tovascular endothelium with liposomes” (projectcoordinator: Manuela Călin).

● 2000: Romanian Academy grant:“Liposome characterization for drugsdelivery”. (project coordinator: Elena Butoi).

● 1999-2001: National Agency forScience, Technology and Innovation(ANSTI) grant: “the expression of celladhesion molecules in valvular endothelium.Involvement in the future treatement ofvalvular diseases”(project coordinator:Ileana Mânduțeanu).

● 1999: Romanian Academy grant,376/1999: “Interaction of liposomes withvascular endothelium” (project coordinator:Manuela Călin).

● 1997-1998: Ministry of research andTechnology grant:“Mechanisms involved inmonocyte adhesion to valvular endothelialcells grown in high glucose concentrations”(project coordinator:Ileana Mânduțeanu).

● 1997: Romanian Academy grant:“Monocyte adhesion to valvular endothelialcells grown in high glucose conditions”(project coordinator:Ileana Mânduțeanu).

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AWARDS

● for Women in Science - L’Oréal-UNESCO Award, 2019 (Monica Țucureanu)

● Innovation Award and gold Diplomaat “euroInvent 11th edition europeanexhibition of Creativity and Innovation2019”(Iași) and Excellence Award at PROINVENT, 2019 (Cluj Napoca) for invention:„Vertical (electro) magnetic separator ofisomagnetic nanoparticles. Inventors:Denisa Ficai, Ioana Ardelelean, Cornelia Ilie,Manuela Călin, Elena-Veleria Fuior, AdrianFifere, Mariana Pinteală, GheorgheFundueanu-Constantin, Anton Ficai, MayaSimionescu, Ecaterina Andronescu.

● Herbert-Berler Prize of Excellence ofthe Romanian Academy of Medical Sciences,2015 [Manuela (Călin) Voinea]

● Romanian Academy Prize in biology“Nicolae Simionescu” for the series ofpapers published on the use of targetednanotherapies for inflammation, 2015[Manuela (Călin) Voinea and Elena (Butoi)Dragomir]

● Constantin Velican Award of theRomanian Society for Cell Biology, 2012[Manuela (Călin) Voinea and Elena (Butoi)Dragomir]

● for Women in Science - L’Oréal-UNESCO Award, 2011 [Elena (Butoi)Dragomir]

● first prize awarded by the RomanianSociety of Cell Biology, 2011 [Manuela(Călin) Voinea]

● Prize for Excellence awarded by theRomanian Medical Association, 2010[Manuela (Călin) Voinea]

● first prize at the National SymposiaVIASAN-CEEX, 2008 [Elena (Butoi)Dragomir]

● Prize for Excellence awarded by theRomanian Society of Cell Biology, 2003[Manuela (Călin) Voinea]

● Agora Diabetologica Prize, of theXXVII National Congress for Diabetes,Nutrition and Metabolic Diseases, 2002[Elena (Butoi) Dragomir]

● European Life Scientist OrganizationPrize, 2002 [Manuela (Călin) Voinea]

● Bio-Rad Laboratories Prize forvaluable research with Bio-Rad equipment,2002 (Ileana Mânduțeanu)

● Daniel Danielopolu Prize of theRomanian Academy, 1994 (Cristina Lupu,Maria Calb)

● Emil Racoviță Prize of the RomanianAcademy, 1993 (Ileana Mânduțeanu)

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