Download Cell mechanics regulation by cytoskeleton dynamics: Combined

MARIESKLODOWSKA-CURIEACTIONS
Co-fundingofregional,nationalandinternationalprogrammes(COFUND)
DOC2AMUPROJECT2017CALLFORAPPLICATIONS
Cellmechanicsregulationbycytoskeletondynamics:
Combinedatomicforceandopticalmicroscopy
(CellMechDyn)
1.DESCRIPTIONOFTHEPHDTHESISPROJECT
1.1OBJECTIVESOFTHEPROJECTBASEDONTHECURRENTSTATEOFTHEART
Mechanical forces are essential in almost all cellular processes. For example, stiffening occurs during cell
division,forcegenerationallowscellmigration,andchangesinthemechanicalmicroenvironmentinducecell
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differentiation
.Thus,biologicalfunctiondeterminesthemechanicalpropertiesoflivingcells.Remodeling
and organizing the cytoskeleton mediate these dynamic changes. The cytoskeleton is a complex network of
filaments that forms the structural scaffold of the cell. Among other proteins, the cytoskeleton is mainly
formed of actin, microtubules and intermediate filaments. While the mechanical properties of individual
filamentsremainmostlyunchanged,itsorganization,assembly,crosslinkingandappliedstressallowthecellto
modulateitsmechanicalresponse.
Twoimportantexamplesinwhichcellmechanicsandcytoskeletonorganizationisessentialarecellmalignancy
and cell migration. In the context of cancer, it has been shown that malignant tumor cells are softer but
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generatehighermechanicalforcesthanbenigncells . Migratingcellsareknowntopolarizeandrestructure
theiractincytoskeletondynamicallytoundergomovement,furthermoretheyaredirectedbytheelasticityof
the microenvironment. However, the interplay between cytoskeleton’s dynamic structure and mechanics is
poorlyunderstood,mainlyduetothelackofavailabletoolscombiningmechanicsandstructuralinformation.
One of the most versatile nanotechniques is atomic force microscopy (AFM), as it allows topographical
imagingwithnanometerresolutionandmechanicalinvestigationofthesamplesurfaceatpiconewtonforces.
ForcemeasurementsoflivingcellsusingAFMconsistinapplyingacontrolledforcethatresultinmeasurable
deformation.Combinedwithitsimagingcapabilities,AFMforcemappingprovidesamapoftheelasticityofthe
cell.Therecentapplicationofhigh-speedAFM(HS-AFM)tocarryoutforcemeasurementsonsinglemolecules
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andcellshaswidenedthedynamicrangeatwhichwecanobservebiologicalprocesses .
Opticalmicroscopyplaysacentralroleincellbiologyasitprovidesapowerfultooltodeterminethestructure
andsupramolecularorganizationofcellorganelles,membraneproteinsand,importantly,thecellcytoskeleton.
Advanced optical methods such as super-resolution techniques provide images with tens of nanometers
resolution,whilepolarizationmicroscopyallowstodeterminelocalorientationalorder.
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Matzke,R.,etal.NatureCellBiology2001,3,(6)
Discher,D.E.,etal.Science2005,310,(5751)
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Dembo,M.;Wang,Y.-L.Biophysicaljournal1999,76,(4)
4
Lekka,M.,etal.EuropeanBiophysicsJournalwithBiophysicsLetters1999,28,(4)
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Guo,M.,etal.Cell2014,158,(4)
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Rico,F.,etal.Science2013,342,(6159)
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Rigato,A.,AMiyagi,SScheuring,FRico.NaturePhysics2017,(inrevision)
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Themaingoalofthisprojectistocorrelatethemechanicalresponseoflivingcellswiththestructureofthe
cytoskeleton of tumor and migrating cells by combining advance atomic force and optical microscopy. The
projectisdividedintwospecificaims:
Specific aim 1. To determine the dynamic structure of the cortical cytoskeleton of benign and malignant
tumor cells and its relation with cells’ viscoelasticity. Our group (BioAFMLab) has recently shown that the
viscoelastic response at high frequencies using high-speed AFM provides a signature of the dynamics of the
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cytoskeleton .Moreover,wehaveshownremarkablydifferentviscoelasticresponseofbenignandmalignant
tumorcellsatshorttimescales.However,thecorrelationbetweenthismechanicalsignatureandthestructure
of the cytoskeleton at high resolution is still unknown. In this specific aim we will use HS-AFM to image the
dynamical structure of the cortical cytoskeleton and to probe the viscoelasticity of tumor cells at different
levels of malignancy. The same cells will be fixed for being analyzed using super-resolution microscopy
(Fresnel).Therequiredequipmentandmethodsarereadilyavailableintheimplicatedlabs.
Specific aim 2. To implement a new adaptive microscope that combines AFM and confocal fluorescence
imaging to simultaneously determine the structure and mechanics of migrating cells. Cellmigration,which
playsaprominentroleincancerinvasion,iselusivetoaconcomitantprobingofmechanicsandcytoskeleton
structure,asmigratingcellsconstantlymoveoutofthefieldofviewoftheAFMorfluorescentmicroscope.To
solve this issue, we will couple an AFM to a custom confocal microscope (already operational at Institut
Fresnel)andusecodesign(optics/AFM/dataprocessing)toadapttheAFMandopticalscanningschemeinreal
time to the trajectory of cells so as to keep them in the center of the imaging field. This will allow
simultaneousmechanicalmappingandcytoskeletalimagingoftheentirecellorjustregionsofinterest.The
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migratingcellishighlypolarized .Thus,knowingthedifferentmechanicalpropertiesofthefrontandrearof
the cell is important to understand this complex and essential biological process. Dr. Rico has built multiple
AFMsystemsduringhisPhDandpostdoctenures.Dr.LeGoffhaslongexperienceinthebiologyofmigrating
cellsandontheuseandimplementationofadvancedopticalmicroscopysystems.
ThisprojectinvolvescollaborationwiththetechnologytransferagencySATTSudEst,whichwillprovideadvice
andexpertiseontheprivatesector.ThestudentwillbeinconstantinteractionwiththeSATTSudEstandwill
perform an internship during the last year of PhD. In addition, Dr. Charafe-Jauffret from the Institut PaoliCalmettesforCancerology,MDpractitionerandresearcherandexpertincancerstemcells,willprovideadvice
andsupervisionregardingthebiologicalandclinicalcontexts.
An internship in the laboratory of Dr. Xavier Trepat from the Institut of Bioengineering of Catalonia in
Barcelona(Spain)willallowthestudenttolearn traction microscopy onlivingcellsandtobeexposedtoan
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internationalenvironment.Dr.Trepatisexpertinthestudyofphysicalforcesduringcollectivecellmigration .
Thisprojectcombinesstate-of-the-arttechnologiessuchashigh-speedatomicforcemicroscopy(onlyavailable
in a few labs worldwide) and polarization confocal and super-resolution microscopy (Pioneered at Fresnel).
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Indeed,theBioAFMLabispioneerintheapplicationofHS-AFMinforcespectroscopymode .Theconceptof
adaptivemicroscopesthatusecodesigntooptimizeimaginginrealtimetothechangingbiologicalsystemisa
new field about to explode due to increasing demands in resolution. The project is interdisciplinary, as it
addresses biological processes (cancer malignancy and migration) from a physics perspective. The project is
feasiblesincealltechniquesareavailableintheinvolvedlabsandthesupervisorshavelongexperienceintheir
respective fields. The expected outcomes of the project will provide a long-term collaboration between the
twogroupsandarepronetoresultin industrial patents.Theexposuretothreedifferent sectors:academic,
industrialandclinicalwillresultinstrongrecruitmentperspectivesofthestudentafterPhDcompletion.
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Ridley,A.J.,etal.Science2003,302,(5651)
Sunyer,R.,etal.Science2016,353,(6304)
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1.2METHODOLOGY
Cellculture.TumorcelllinesfrombreastcancerwillbegrownonHS-AFMglasssupportspresentingagridfor
later localization of the same cells using super-resolution microscopy. Cells for migration experiments with
fluorescentactincytoskeletonarealreadyavailableatFresnel.Micropatternsfeaturinglongrailswillbeuseto
directcellmigrationalongasingledirection.
HS-AFM.TheBioAFMLabhastwoHS-AFMsystems(Ando-typeRIBM,Japan)andaconventional/cellularAFM
(JPK,Germany)coupledtoopticalmicroscopy.HS-AFMimagingwillbecarriedoutusingfastcantileversathigh
amplitude tapping mode using a wide scanner. Viscoelastic measurements at high frequencies using HS-AFM
will be carried out using cantilevers with a grown electron beam deposited tip featuring a sphere of ~1µm.
Measurementswillconsistonapplyinglowamplitudesinusoidaloscillationsaroundaconstantindentationat
frequencies between 1Hz and 100kHz. The HS-AFM systems are already operational for the proposed
measurements.
Optical microscopy. For correlative HS-AFM/Fluorescence microscopy, after mechanical measurements, cells
willbefixedandstainedforopticalimaging.Toprobespatialorganizationoftheactincytoskeleton,cellswill
beimagedwithsuperresolutionmicroscopy,firstonacommercialSTEDmicroscopeoftheFrancebioimaging
platform at IBDM near the BioAFMLab, then using STORM at Institut Fresnel (higher resolution but more
complexanalysisthanSTED).Furtherstructuralinformationontheorientationalorderoftheactinmeshwork
willbegainedusingpolarizationconfocalmicroscopy,andpolarizationSTORMmicroscopyatFresnel.
AFM-basedsystemcoupledtoconfocalmicroscopy.TheAFMsystemwillbebasedonadesignpreviouslybuilt
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byDr.Rico .Thesystemwillfeaturez-piezoforcantilevermovementandxy-piezosforsamplemovement,all
mountedonthestageofamotorizedopticalmicroscope.PrototypingwillbedoneattheBio-AFMlab,while
thefinaldesignwillbebuiltattheworkshopofInstitutFresnelandcoupledtoaconfocalmicroscope(already
operational). As force mapping is slower than confocal imaging, an optical image-based feedback will ensure
repositioningofcellsinthefieldofviewoftheAFMand“guide”themechanicalmappingtospecificregionsof
interestsuchasthefrontandrearofmovingcellsidentifiedbyimageanalysis.Softwarewillbedevelopedin
LabView(NI,France)tocombinetheAFM-basedsysteminforcemappingmodeandthetrackingofmigrating
cellsusingconfocalmicroscopy.ThegroupatFresnelinstitutehasalreadydevelopeddifferentscanningcontrol
softwarepackagesthatwillbeeasilymodifiedtoallowforcemapping.Inparticulartherecentimplementation
of“smartscans”byDr.LeGoffthatusesimageanalysistofocuslaserscanningtoregionsofinterestofhigh
informationcontentwillbeeasilyconvertedforthe“smart”mechanicalmappingaforementioned.
1.3WORKPLAN
Theprojectwillbedividedintotwomainpartsrelatedtothespecificaims.Thefirststagewillbedevotedto
theapplicationofHS-AFMforimagingandviscoelasticmeasurementsofthecorticalcytoskeletonoftumor
cells(BioAFMLab)andtheposteriorimagingoffixedcellsusingsuper-resolutionimaging(Fresnel).Thiswill
allowthestudenttomastertheuseofHS-AFMtechnologybothinimagingandforcemodesandfamiliarize
withadvancedopticalmicroscopymethodsandinstrumentation.Thestageintheinternationalpartnerwillbe
partofthisspecificaim.Thisstageisdividedinto6tasks:
Task1.1
Task1.2
Task1.3
Task1.4
Task1.5
EstablishingcellcultureconditionsforHS-AFMandposteriorsuper-resolution(months1-9)
Microrheologymeasurementsontumourcells(months3-9)
HS-AFMimagingofcorticalcytoskeletonoftumourcells(months3-12)
Imagingoffixedtumourcellswithsuper-resolutionopticalmicroscopy(months6-12)
Internationalinternship(months12-15)
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Chu,C.,Celik,E.;Rico,F.;Moy,V.Tetal.PLoSONE2013,8,(5)
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Task1.6 Dataprocessing,correlationandanalysis.Manuscriptwriting(months12-18)
Theexpertisegainedduringthefirststagewillbeessentialforthesecondstageoftheprojectdedicatedto
implementandAFM-basedsystemonaconfocalmicroscopetostudymigratingcells.Thissecondstagewillbe
devotedtothedesignandconstructionoftheAFM-basedsystemanditstestingonlivingcells,itscouplingon
thestageofthespinningdiskconfocalmicroscopeandfinallyapplicationtothestudyoflivingmigratingcells.
Theinternshipintheprivatesectorpartnerwillbepartofthisstage.Itisdividedinto5tasksplusthesiswriting:
Task2.1 Design,constructionandtestingoftheAFM-basedsystem(months15-21)
Task2.2 CouplingoftheAFM-basedsystemontheconfocalmicroscope(months18-21)
Task2.3 Simultaneousmechanicalmappingandconfocalimagingofmigratingcells(months21-27)
Task2.4 Dataprocessing,correlationandanalysis.Manuscriptwriting(months28-33)
Task2.5 Thesiswriting(months30-36)
Tasks1.1-1.3and2.1willbecarriedoutintheBioAFMlab,tasks1.4,2.2and2.3willbecarriedoutatthe
Fresnelinstitute.Tasks1.6and2.4willbecarriedoutbetweenthetwolabs.Anintersectorialinternshipat
SATTSudEstwillalsotakeplaceinthecourseofthelastyearofthePhD.
1.4SUPERVISORSANDRESEARCHGROUPSDESCRIPTION
ThestudentwillbecosupervisedbyFelixRico(Bio-AFM-Lab)andLoicLeGoff(Fresnel)
Mainsupervisor:Dr.FelixRICOismaîtredeconférenceswithChaireMixteattheBio-AFM-Lab,U1006INSERM
&AMU.SinceJanuary2017,heisinterimdirectoroftheunit.HewilldefendhisHDRon17January,2017.He
holdsaPhDinbiophysicsfromtheSchoolofMedicineoftheUniversityofBarcelona,Spain.Hehaspublished
35articlesandreviewsinjournalslikeScience,NanoLettersandtheBiophysicalJournal.Hehascontributedto
about36conferences(13asspeaker)andhasbeeninvitedtoabout24seminars.Hehassupervisedmasterand
PhDstudents,postdocsandvisitorresearchers.HeteachesatAMU(atundergraduateandgraduatelevels)and
isregularlectureratinternationalschools(e.g.AFMBioMed2011-2016,CNRSthematicschools).Dr.Ricohas
been working in force spectroscopy with AFM since 2001 when he started his PhD on cell mechanics. His
postdocinthelaboratoryofVincentT.Moy(Miami,FL)focusedonthebiophysicaldeterminantsofadhesion
molecules. At Institut Curie, Dr. Rico gained further expertise in force spectroscopy to probe cellular and
molecular mechanics, and high-resolution imaging and force mapping on native membranes and cells. He
recentlypioneeredtheimplementationofthefirsthigh-speedforcespectroscopysystem,appliedittounfold
singleproteins.Relevanttothisproject,asco-directorofA.Rigato’sthesis,hedevelopedmechanicalmapping
on micropatterned cells and pioneered the implementation of high-speed microrheology measurements on
livingcells(inrevision).HewasawardedanANRJCJCgrant(260k€)in2015andhasanannualassignmentof
10k€/yearlinkedtohischairemixteposition.Partofthesefundswillbeavailabletosupportthisproject.
Co-supervisor: Dr. Loic Le Goff: DrLoïcLeGoffischargéderecherchéatCNRS.AfteraPhDatInstitutCurie
(Paris) on actin filaments dynamics and a postdoc at Rockefeller University (New York) on the biophysics of
hearing,hegainedinterestindevelopmentalbiologyuponjoiningtheInstitutdeBiologieduDévelopementde
Marseille.Herecentlyuncoveredtheimportanceofglobalpatternsofmechanicalstressinshapingthegrowing
D. melanogaster wing. He also pioneered in vivo imaging for D. melanogaster wings and developed tools to
quantify deformations therein. At Institut Fresnel, in the MOSAIC team, he further develops new imaging
endeavors on tissues while pursuing his interest on cell and developmental biology. Of particular interest to
him are processes of guided cell migration in embryos, as well as the development of new adaptive
microscopes that can track and optimally image migrating cells over large distances. The development of
adaptive microscopes, upon which aim 2 builds, is financially supported by the Institut Carnot Star and the
transfer agency SATT SudEst. The onsite expertise in super resolution microscopy, polarization resolved
microscopyandimageanalysiswillbeinstrumentalfortheachievementofthisPhD.
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2.3IDIMENSIONSANDOTHERASPECTSOFTHEPROJECT
2.1 INTERDISCIPLINARYDIMENSION
Theinterdisciplinaritydimensionwillbefirstlyassuredbythecandidatebeingenrolledintwodoctoralschools:
ED062(SciencesdelaVieetdelaSanté)andED352(PhysiqueetSciencesdelaMatière).Theprojectcovers
two of the priority thematic axes of the DOC2AMU program: nano-health and imaging. The supervisors are
formedunderaninterdisciplinaryperspective,asbothusephysicalapproachestoaddressbiologicalquestions.
The biological experience is however different, Rico has a molecular to cellular level expertise, while Le Goff
has been devoted to cells and organisms. The PhD project clearly combines various disciplines. First, physics
andcellbiology:thesystemsofstudyinvolvetwoimportantbiologicalprocessesmalignancyincancerandcell
migration,whilethemainapproachisphysical,beingthusoriginal.Andsecond,thestudentwillgainexpertise
in two different techniques, advanced optical microscopy and imaging and mechanical measurements using
high-speed andconventional atomic force microscopy.Masteringdifferenttechniquesisessentialforfuture
research in the field of biophysics. The experience and collaboration established from the PhD will have two
mainbenefits:theFresnelinstitute,anadvancedopticscenter,willgaininsightintoatomicforcemicroscopy
and biophysics of cancer cells. The U1006, an AFM lab, will gain access and knowhow on advanced optical
microscopyandoncellmigration.Itisexpectedthatthiscombinationofmutualbenefitwillallowestablishing
along-termcollaborationbetweenthetwogroups.
2.2INTERSECTORIALDIMENSION:
TheintersectorialdimensionissecuredbyourcollaborationwiththetechnologytransferagencySATTSudEst.
The SATT SudEst has already supported us financially for the development of codesign optical microscopy,
whereimageanalysisanddataacquisitionareintermixedinordertooptimizetheimagingprocess.Thecurrent
projectextendstheconceptofcodesigntotheAFM/optics/dataanalysismodality.Itbearsgreatpromisesfor
thedevelopmentofanewcuttingedgetechnologywithastrongpotentialityforcommercialization.Thetwo
supervisors and the PhD student will be in constant interaction with the SATT SudEst for advise and support
regarding intellectual property, patents and contacts with companies interested in this development. The
studentwillperformaninternshipattheSATTSudEst,duringwhichhe/shewilllearnmoredeeplyconcepts
related to intellectual property and make contacts with other technology actors from the private sector
(Optitec,smallbusinessandlargermicroscopycompanies,seetheletterofsupportfromSATTSudEst).
Besidesitsrootsinfundamentalbioscience,connectionofthePhDworktohealthsectorwillbeensuredbyour
regional collaboration with a practitioner in cancerology, Dr. Charafe-Jauffret from Insitut Paoli-Calmettes.
The student will interact with hospital technicians to learn the conventional cancer diagnosis and prognosis
toolsusedintheclinic,providingexperienceinthecommonprocedures,possibleproblemsandcurrentneeds.
2.2 INTERNATIONALDIMENSION:
Theinternationaldimensionwillbefulfilledbyatwo-monthsecondmentduringthesecondyearofPhDinthe
laboratory of Dr. Xavier Trepat at the Institut of Bioengineering of Catalonia in Barcelona, Spain. Dr. Trepat
expert in traction microscopy applied to collective cell migration. This secondment will complement the
mechanical AFM data by measuring traction forces generated by the tumor cells studied in aim 1. Traction
microscopyconsistsofanelasticsubstrateofknownstiffnesswithembeddedfluorescentbeads.Cellsgrowing
onthegelgeneratetractionforcesthatdeformthegel.Trackingthepositionofthebeadsprovidesameasure
ofthedeformationofthegel,whichtranslatesintocellgeneratedtractionforces.Tractionmicroscopywillbe
laterappliedtomigratingcellsinthecontextofaim2.Apartfromtheinternship,thestudentwillattendone
internationalconferenceperyear(BiophysicalSocietyMeeting,InternationalMicroscopycongress…).
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3.RECENTPUBLICATIONS
High-speedforcespectroscopyunfoldstitinatthevelocityofmoleculardynamicssimulations.Science,2013,
342(6159):741-743.FelixRico,LauraGonzalez,IgnacioCasuso,ManelPuig&SimonScheuring
Ahybridhigh-speedatomicforce-opticalmicroscopeforvisualizingsinglemembraneproteinsoneukaryotic
cells.NatureCommunications,2013,4:2155,AdaiColom#,IgnacioCasuso#,FelixRico&SimonScheuring
AglobalpatternofmechanicalstresspolarizescelldivisionsandcellshapeinthegrowingDrosophilawing
disc.LeGoff,L.,Rouault,H.&Lecuit,T.Development(2013).
Dynamicclonalanalysisbasedonchronicinvivoimagingallowsmultiscalequantificationofgrowthinthe
DrosophilawingdiscHeemskerk,I.,Lecuit,T.&LeGoff,L..Development.(2014).
Atomic Force Microscopy Mechanical Mapping of Micro patterned Cells Shows Adhesion Geometry-
DependentMechanicalResponseonLocalandGlobalScales.RigatoA,RicoF,EghiaianF,PielM,ScheuringS.
ACSNano.2015Jun23;9(6):5846-56.
Structural, mechanical, and dynamical variability of the actin cortex in living cells. Eghiaian F, Rigato A,
ScheuringS.BiophysJ.2015Mar24;108(6):1330-40.
BookChapter
Atomic force microscopy studies of the mechanical properties of living cells.RicoF,EPWojcikieviczandVT
Moy,inAppliedScanningProbeMethodsIX,B.Bhushan,H.Fuchs,M.TomitoriEds.,Springer,NewYork,2008
Mechanicalforcesandgrowthinanimaltissues.LeGoff,L.&Lecuit,T.ColdSpringHarb.Perspect.Biol.(2016).
4.EXPECTEDPROFILEOFTHECANDIDATE
ThecandidateshouldholdaMasterdegree(orequivalent)inafieldrelatedtothesubjectofstudy(biophysics,
biomedicalengineering,biologyorsimilar).Experienceinopticalmicroscopyand/oratomicforcemicroscopy
appliedtobiologicalsystemsandcellcultureskillswillbeuseful.Theprimarymethodofresearchusedinthis
project(HS-AFM)requiresdetailedknowledgeofphysicstooperateit,sothecandidateshouldbefamiliarwith
physicalconcepts related to scanning probe microscopies. Transnational mobility is an eligibility criterion for
receivingMarieSkłodowska-CurieCOFUNDfunding:thecandidatemustnothaveresidedorcarriedouttheir
main activity (work, studies, etc.) in France for more than 12 months in the 3 years immediately prior to
thereferencedate.GoodknowledgeofEnglishisessential.Thecandidateshouldbeopen-minded,analytical,
dedicatedandmotivated.Thecandidatemustalsobereadytospendlongerperiodsoftimeintheinstitutions
oftheinternationalandintersectoralpartners.
5.SUPERVISORS’PROFILES
FelixRicoprofessionalprofile
2001-2006 PhDinBiophysics,UniversityofBarcelonaSchoolofMedicine,Spain
2006-2008 FulbrightPostdoctoralFellow,VincentT.Moy’slab,,UniversityofMiami,FL,USA
2009-2011 MarieCuriePostdoctoralFellow,UMR168,InstitutCurie,Paris
2012-2013 ResearchAssociate,U1006INSERM&Aix-MarseilleUniversité,Marseille
2013-now
Maîtredeconférences/Interimdirector,U1006INSERM&Aix-MarseilleUniversité,Marseille
FormerPhDstudentsofthemainsupervisorFelixRICOandpublications
•AnnafrancescaRIGATO,Marseille,France(03/2013-11/2015PhDthesis,12/2015–nowPostdoctobe)
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High-frequencymicrorheologyrevealscytoskeletondynamicsinlivingcells
RigatoA,AMiyagi,SScheuring,andFRico*.NaturePhysics,inrevision
AtomicForceMicroscopyMechanicalMappingofMicropatternedCellsShowsAdhesionGeometry-
DependentMechanicalResponseonLocalandGlobalScales
RigatoA,RicoF,EghiaianF,PielM,ScheuringS.ACSNano.9(6):5846-56.2015
Structural,mechanical,anddynamicalvariabilityoftheactincortexinlivingcells
EghiaianF,RigatoA,ScheuringS.BiophysJ.Mar24;108(6):1330-40.2015
Mechanicsofproteinswithafocusonatomicforcemicroscopy
RicoF,RigatoA,PicasL,ScheuringS.JNanobiotechnology.11Suppl1:S3.2013
•CalvinCHU,Miami,FL,USA(07/2006-12/2008PhDthesis,03/2011–nowinindustry)
Elongatedmembranetethers,individuallyanchoredbyhighaffinityα4β1/VCAM-1complexes,arethe
quantalunitsofmonocytearrests
ChuC,FRico,ECelik,andVTMoy.PLOSOne.8(5)e641872013
Temperaturedependenceofintegrin-mediatedcelladhesion
RicoF*,CChu,MHAbdulreda,YQinandVTMoy.BiophysJ99(5)1387-13962010
Force-clampmeasurementsofreceptor-ligandinteractions
RicoF,CChuandVTMoy.MethodsMolecularBiology.736331-532011
LoicLeGoffprofessionalprofile
2002: PhD,InstitutCurie,Paris
2002-2006:Post-doc,HearingResearch-Supervisor:AJHudspethTheRockefellerUniversity,USA
2007-2015:CNRSResearcher(section22)InstitutdeBiologieduDéveloppement,Marseille
2015- :CNRSResearcher(section22:cellanddevelopmentalbiology)InstitutFresnel,Marseille
Mentoredpostdocs
•HervéRouault,IBDM,Marseille12monthsofco-supervisionin2011
AglobalpatternofmechanicalstresspolarizescelldivisionsandcellshapeinthegrowingDrosophilawing
disc.LeGoff,L.,Rouault,H.&Lecuit,T.Development(2013).
•IdseHeeskerk,KITP,SantaBarbara,USA.9monthsofsupervisionin2013
Dynamic clonal analysis based on chronic in vivo imaging allows multiscalequantification of growth in the
DrosophilawingdiscHeemskerk,I.,Lecuit,T.&LeGoff,L..Development.(2014).
OnecurrentPhDStudent:HaiNguyenPhDStartedon6/01/2017
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