AlfredCuschieriInstituteforMedicalScienceandTechnology

DivisionofSurgeryandCancerUniversityofDundee&

SchoolofmedicalsciencesScuolaSuperioreS’Anna,Pisa

NANO-TECHNOLOGYFORHEALTH

NANOTECHNOLOGYMARKETS

• Nanotechnologymultidisciplinaryfieldthatcoversdiversearrayoftechnologiesderivedfromengineering,physics,chemistry,materialsscienceandbiology

• Burgeoningfieldhasledtorapidadvancesintechnologyandlifeandphysicalsciences

• UsheredneweraofNano-Medicinewithnanomaterialsdesignedtointeractwiththebodyatsubcellularmolecularscaleswithahighdegreeofspecificity

• Potentialfortargetedcellularandtissue-specificclinicalapplicationsdesignedtoachievemaximaltherapeuticefficacywithlesssideeffects

NANOTECHNOLOGY

• Onenanometre(nm)=onethousandmillionthofameter• Forcomparison,asinglehumanhair=80,000nmandanRBC=7,000

nm• Definitionofnanoscale:from100nmdowntothesizeofatoms

(approximately0.2nm)

• Atthisscalethepropertiesofmaterialsareverydifferentfromthose

atalargerscale

NANOSCALE

UNIQUEPROPERTIESOFMATERIALSATTHENANOSCALE

• Nanomaterialshaverelativelylargersurfaceareawhencompared

tobulkmaterials- morechemicallyreactive- somematerialsare

inertandbecomereactiveintheirnanoscale

• Nanoscaleaffectsthestrengthand/orelectricalproperties

• Quantumeffectsbegintodominatethebehaviourofmaterials

affectingtheiroptical,electricalandmagneticproperties

NANOTECHNOLOGY&NANOSCIENCE: STUDYOFMATTERATANATOMICANDMOLECULARSCALE

RichardFeynmanin1959:atnanoscalethegravitywouldbecomelessimportant,surfacetensionandvanderWaalsattractionpredominate– highlyreactivequantumeffects

• Nanotechnologyindrugdelivery• Nanotechnologyingenedelivery• MRIcontrastandmolecularimaging• Antibacterialagents• Nano-fluidicdiagnosticchips• Nano-formulationsofestablishedchemotherapeuticanti-canceragents• Nanohyperthermicablation• Controlofcellmovement:regenerationandregenerativemedicine• Nano-molecular imprintingpolymerstoreplacemonoclonalantibodies• Nano-piezoelectric transducers– therapiesbasedonelectricstimulation

NANOMEDICINE

• Becauseoftheirsustained-releasecharacteristics

• Biocompatibility

• Biodegradability

• AbilitytoprotectDNAfromdegradationinendolysosomes

PGLAandPLANANOPARTICLESforDRUGDELIVERYandasGENETHERAPYVECTORS

• PolymericNPsofPGLAareabletoescaperapidlyfromtheendolysosomal

compartmenttothecytoplasmfollowingintracellularuptakeandthusthe

carrier-DNAcomplexescapelysosomaldigestion

• ThetransfectionyieldcorrelatesnegativelywithsizeofPGLAPLAparticles

suchthat100nmparticlesproducethehighesttransfectionrate

• Thehighertransfectionefficiencyofthesmallerfractionnotrelatedtothe

differencesintheDNAloading,cellularuptake,orDNAsrelease

NON-VIRALTRANSFECTIONbyPGLANP

• QDswhichrelyonbioluminescenceresonanceenergytransfer- abletoconvertchemicalintophotonicenergy,withgreatlyincreasedfluorophoreexcitationandreductionintheeffectsoftissueautofluorescence

• NeedforbiocompatibleQDstoovercometheinnatetoxicityofheavy

metalQDswhichrequirecomplexpolymerencapsulationfor

biocompatibility

ADVANCESINQUANTUMDOTS

• Nanotechnologiesenabledintracellularimagingthroughattachmentof

QDsorsyntheticchromophorestoselectedmoleculesorbythe

incorporationofnaturalfluorescentproteinswithopticaltechniques

suchasconfocalmicroscopy

• Nanosensorsbasedonfluorescenceresonanceenergytransfer(FRET)

whichusesQDslinkedtoDNAprobes.Inthissystem,afluorescence

resonanceenergytransferdonor-acceptorensembleformsasaresult

ofthebindingofthetargetstrandtoadyelabelledreporterstrand

NANOTECHNOLOGYFORIMAGING

CELLULARIMAGINGbyQDs

Human breast cancer cells tagged with quantum dots

Fluorescence microscopy of labelled chromosomes

• Smallsphericalartificialvesiclesproducedfromnaturalnontoxic

phospholipidsandcholesterol

• Liposomesareparticularlyusefulasgenetherapyvectorsbecauseof

theirabilitytopassthroughlipidcellmembrane

• Likelytoreplaceviralvectors,especiallycationicliposomes

• Formcomplexes(lipoplexes)andareincreasinglypreferredforthe

intracellulardeliveryofreporterortherapeuticgenesbecauseof

hightransfectionrates

LIPOSOMES

• Dendrimers- macromolecularcompoundsconsistingofaseriesofbranchesaroundaninnercore

• DendriticpolymersareeffectiveasDNAconjugates

• Resultingdendrimer-DNAcomplexdiffersfromencapsulationinasthemechanismresponsibleforgeneretentionconsistsofelectrostaticinteractionsbetweenthenegativelychargedphosphateDNAgroupsandthepositivelychargedaminogroupsonthepolymer

• Polyamidoamines(PAMAMs)arethemostoftenusedandeffective

• dendrimersforgenedelivery

DENDRIMERS

EUNANOTECHNOLOGYPLATFORM

EUDefinitionofNanomedicine:

Nanomedicineisdefinedastheapplicationofnanotechnologyto

achievebreakthroughsinhealthcare.Itexploitstheimprovedand

oftennovelphysical,chemicalandbiologicalpropertiesofmaterials

atthenanometerscale.Nanomedicinehasthepotentialtoenable

earlydetectionandprevention,andtoessentiallyimprovediagnosis,

treatmentandfollow-upofdiseases

• Nanomedicineshouldaimformeaningfulimprovementsinareasthat

containthemostseverechallengesinfuturehealthcare

• Sixdiseaseareasselectedbasedonthefollowingcriteria:diseasesthatreduceQoL

highprevalence

imposeahighsocio-economicalburdenonsociety

• nanotechnologypredictedtohavehighimpactonthecareprocessfor:

Cardiovasculardisease,Cancer,Musculoskeletaldisorders,

NeurodegenerativeandPsychiatricdisorders,Diabetes,Bacterialandviral

infections

EUNANOTECHNOLOGYPLATFORM

• Controlleddrugdelivery

• Targeteddrugdelivery

• Reduceddrugtoxicity

• Theranosticdiseasemanagement

POTENTIALofNANOTECHNOLOGYINDRUGTHERAPY

• Nanochipsandnanoarrays• Constructedwithmicroelectromechanicalsystem(MEMS)• Nanotechnologyisbeingusedtoovercomesomeofthelimitationsof

biochiptechnology• Biologicaltestsmeasuringthepresenceoractivityofselected

substancesaremadequicker,moresensitive,andmoreflexiblewhencertainnanoscalereagentparticlesareused

• Detectionofspecificbiomolecularinteractions• Magneticnanoparticles,boundtoasuitableplasticnano-antibodies

areusedtolabelspecificmolecules

NANOTECHNOLOGYFORMOLECULARDIAGNOSTICSNANOCHIPSvsBIOCHIPS

• Nanotechnology‘plasticantibodies’onachipisanewparadigmfor

totalchemicalanalysissystems

• Theabilitytomakechemicalandbiologicalinformationmuchcheaper

andeasiertoobtainisexpectedtofundamentallychange:

healthcare

foodsafety

crimedetection/lawenforcement

POTENTIALofNANOTECHNOLOGYonaCHIP

Incidenceofcardiacevents CongestiveHeartfailure

Freedoxorubicin 29% 8%

Myocet 13% 2%

NANO-ONCOLOGY

JournalofNanomaterialsVolume2011(2011),Article ID164506,http://dx.doi.org/10.1155/2011/164506 CRiggio,EPagni,VRaffa,ACuschieri

Table 3:Valuesforhalf-lifeandAUCfortwoliposomal

formulationsandfreedoxorubicin.

Half-lifetime AUC(AreaUnderCurve)

Freedoxorubicin 0.2 h 4 μgh/mLMyocet 2.5 h 45 μgh/mLCaelyx/Doxil 55 h 900 μgh/mL

NANO-ONCOLOGYwithLIPOSOMALFORMULATIONS

JournalofNanomaterialsVolume2011(2011),Article ID164506,http://dx.doi.org/10.1155/2011/164506 CRiggio,EPagni,VaRaffa,ACuschieri

WIRELESSELECTROSTIMULATIONTHERAPYBYPIEZO-ELECTRICNANOSTRUCTUREDIMPLANT

video

Deepbrainstimulation(DBS)ofsubthalamicnucleusforParkinson’sdiseaseD

Stereotacticinjection

n-MOLECULARIMPRINTINGPOLYMERSSOLUBLEPLASTICANTIBODIES

HEL machine for ‘thin film’ nano-MIPs@IMSaT – University of Dundee SchematicrepresentationoftheautomatedsynthesisofnanoMIPs

usinganimmobilisedtemplate

BIOLOGICS THERAPY with n-MIP PLASTIC ANTIBODY against EGFR EQUIVALENT TO HERCEPTIN

extracellularandtransmembranedomaininterfacesinepidermalgrowthfactorreceptorsignalling

CONFOCALMICROSCOPYSTUDIESOFTUMOURCELLSWITHPEGYLATEDn-MIPS

ConfocalmicroscopyofHaCaTcellStreatedwith2.5nMPEGylatedn-MIPs.Stainingagents:DAPI(blue)forthenucleus,DIO(green)forthecytoplasm,andRhodamine(red)fornanoMIPs

HYBRIDQD-nMIPagainstVEGFR

ZEBRAFISHEMBRYOTUMOURXENOGRAFTSTUDIESwithQD-MIPIMPRINTEDagainsthVEGF

WM-266 cells

QD-MIPs imprinted against hVEGFinjected in WM-266 zebrafish embryos

Embryo#1

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CONTROL- QD-NIPimprintedagainstvancomycin

WM-266 cells

QD-NIPs imprinted against vancomycininjected in WM-266 zebrafish embryos

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CONTROL– XENOGRAFTwithTUMOURCELLSnotexpressingVEGFR

A375 cells no VEGF expression

QD-MIPs imprinted against hVEGFinjected in A375 zebrafish embryos

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HOMINGofanti-VEGFRQDn-MIPONLYtoXENOGRAFTS

EXPRESSINGVEGFR(266cells)

Mean of the distances nanoprobe-cellsin zebrafish embryos injected with either WM-266

or A375 cells

QD-MIP W

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QD-nip WM-26

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QD-MIP A37

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EGFRn-MIPsbindingtolungandbreastcancercells

LOWCOSTBIOLOGICS THERAPYWITHnMIPs

n-MIPsvs.MONOCLONALANTIBODIES

• Majorresistancetointroductionofn-MIPsincancertherapyand

fromsomeofthefundingbodies

• Difficulttodefendbecauseoftheobviousadvantagesofn-MIPsovermAbs:Farcheapertoproduce- greatlyreducedtherapeuticcoststoNHSLongshelflifeGreatlyimprovedbiodistributionLikelyoralbio-availabilityNoallergicissues

• Althoughsomeconcernsmaybeill-founded,thetoxicologyofmanynanomaterialshasnotyetbeenfullyevaluated

• EUCompaniesareparticipatingintheEuropeanNanosafeConsortium- evaluatesthepossiblerisksfrommanufactureanduseofnanomaterials

• Wheninhalednanomaterialsreachthelungswithsubsequentpossiblespreadtootherorgans

• Atthecellularlevel,abilitytoactasagenevectorhasbeendemonstratedforNPs

• ExperimentalevidencewhichdemonstratesthatDNAcanwraparoundCNT,raisingsomeconcernsoveradverseconsequencesofCNTsafterenteringthehumanbody

RISKofn-MATERIALStoHUMANHEALTH

• Nanomedicinebasedonnanoscience/engineering,biologyandmaterialscience

hasthepotentialtoimprovehealthcare

• Itwillresultinmoreeffectivetherapiesforawidespectrumofhumandisorders:

genetic,commonlifethreateningandneurodegenerative

• Ushernewapproachestocurrentlyuntreatabledisorders

• Leadtocheaperandmoreeffectivepharmaceuticalswithreducedsideeffects

• Needforrobustscientificstudiesonthetoxicityandharzardsonnanomaterials

andtheadoptionofrequiredprotectivemeasures

CONCLUSIONS

• Colloidalmicroparticles>100nm– 1000nm• Nanoparticlesarecolloidalparticles<100nm• Aptameres4- 10nmsize:DNA,RNA,andpeptideaptamersfunctionas

direct‘antibodiesagainstanymoleculartarget- smallmolecules,toxins,peptides,proteins,viruses,bacteriaetc.

• Aptamersexhibithighspecificityandaffinitystrongbindingtotheirspecifictargets- theystructurallyconformtobindtotheirtargets,whencomparedwithantibodies,whichrequireantigensandepitopes

• Bindingofaptamertospecifictargetdoesnotcauseanimmuneresponse

MICROPARTICLES,NANOPARTICLES,APTAMERS