Québec Highlights › wp-content › uploads › 2019 › 12 › Quebec.pdf• CLEC-2, the PDPN receptor, is expressed on normal platelets and was found to be necessary for the separation

QuébecHighlights

Denis Claude Roy, MD

Director of Research, East Montreal andCentre of Excellence in Cell TherapyHôpital Maisonneuve-RosemontCEO, CellCAN: Regenerative Medicine and Cell Therapy NetworkProf. Medicine, Université de Montréal

TranscriptionalLandscapeofAPLIdentifiesAberrantPodoplanin ExpressionAsaDefiningFeatureandMissingLinkfortheBleedingDisorderofThisDiseaseVincent-PhilippeLavallée,M.D.1,2,MiriamMarquis,Ph.D.3*,Marie-ÈveBordeleau,Ph.D.2*, Jalila Chagraoui,Ph.D.4*, TaraMacRae,M.Sc.4*, IsabelBoivin,M.Sc2*,GenevièveBoucher,M.Sc2*, PatrickGendron,M.Sc2*,SébastienLemieux,Ph.D.5,6*, ArnaudBonnefoy,Ph.D.7,8*,GeorgesE.Rivard,M.D.7,8,JoséeHébert,M.D.1,2,3,8 andGuySauvageau,M.D.,Ph.D1Hematology-OncologyDivision,Maisonneuve-RosemontHospital,Montréal,QC,Canada2TheLeucegene project atInstituteforResearchinImmunology andCancer,UniversitédeMontréal,Montréal,QC,Canada3QuebecLeukemia Cell Bank,Montréal,QC,Canada4InstituteforResearchinImmunology andCancer,Montréal,QC,Canada5InstituteforResearchinImmunology andCancer,UniversitédeMontréal,Montréal,QC,Canada6DepartmentofComputerScienceandOperationsResearch,UniversitédeMontréal,Montréal,QC,Canada7CHUSainte-Justine,M

Monday,December 5,2016:4:30PMMarriottGrand2-4(MarriottMarquisSanDiegoMarina)

TranscriptionalLandscapeofAPLIdentifiesAberrantPodoplanin ExpressionAsaDefiningFeatureandMissingLinkfortheBleedingDisorderofThisDiseaseVincent-PhilippeLavallée

• Acutepromyelocytic leukemia (APL)isafavorable-risksubgroupofAMLcharacterizedbythet(15;17)translocation.

• TheleadingcauseofearlydeathinAPLisuncontrolledbleedingmostlyattributedtoaberrantexpressionoftissuefactor(F3)andannexin A2(ANXA2)onleukemicpromyelocytes leading todisseminated intravascularcoagulationandhyperfibrinolysis,respectively.

• Topreventortreatsuchcomplications,early suspicionofAPLandrapidinitiationoftherapyandsupportivemeasuresarecritical.

• Podoplanin orPDPNisasurfaceglycoproteinexpressed inmostcell types,butnotinbloodcells.• CLEC-2,thePDPNreceptor,isexpressedonnormalplateletsandwasfoundtobenecessary for

theseparationofbloodandlymphaticvesselsduringembryogenesis.• PDPNexpression(whetherendogenousorectopic)incell lines inducesplateletaggregation,

whichcanbeinhibitedbychemical toolcompoundsorbymonoclonalantibodies

• AimsandMethods: Analysisofthetranscriptomeof30APLcomprisedintheLeucegene 430AMLcohort.


• Results: Severalmutatedgenesinthiscohort,mostofwhicharenon-specificandpreviously identified.

• CEBPE mutationsweretheonlyexceptionandwerespecifictoAPLspecimensinthiscohort (2/30vs0/400,p=0.005).

• Authorsidentified PDPN asthesinglemostdifferentiallyoverexpressedgeneinAPL

• PDPN isnotexpressedinwholeblood,bonemarrowandinanysortedcellsubpopulations fromthesenormaltissues,includingpromyelocytes.ThisindicatesthatplateletsareneverexposedtoPDPNintheadultvasculatureandrevealsthatthisgeneisectopicallyexpressedinAPLpromyelocytes.

• Hypothesis:aberrantPDPNexpressiononleukemicpromyelocytes contributestoabnormalplateletaggregationinAPLpatients.High PDPN expressionisassociatedwithlowerplateletcountsatpresentation(18vs34x1012/L,medianPDPN expression≥10vs<10RPKM,p=0.016,FigC).

• Stronginversecorrelationwasobservedbetweenthenumberofestimatedcirculating PDPN+ promyelocytes andplateletcounts

• Byincorporatinganti-PDPNantibody intheEuroFlow protocol,PDPNexpressiontestwas90%sensitiveand100%specificforAPL(n=48and50APLandnon-APLprimaryAML,respectively).Ofnote,5APLcasesconsideredpositiveexpressedlowlevelsofPDPN.

• ComparingexpressionofallcoagulationandfibrinolysisgenesinAPL(n=30)tothatofnon-APLspecimens(n=400),PDPNwasthemostdiscriminatorytranscript.ThisresultstandsinsharpcontrastwiththatfoundwithF3andANXA2whichlargelyoverlapintheseAPLversusnon-APLhumanAML.

769 Chemo-TranscriptomicAnalysisofComplexKaryotypeAMLRevealsIncreasedExpressionofCellCycleComponents andExquisiteDependencyonPolo-likeKinase1

Vincent-PhilippeLavallée,M.D.1,2,ClarisseThiollier,Ph.D.2*, CélineMoison,Ph.D.2*,Marie-ÈveBordeleau,Ph.D.2*,IsabelBoivin,M.Sc2*,GenevièveBoucher,M.Sc2*,PatrickGendron,M.Sc2*,SébastienLemieux,Ph.D.3,4*, AnneMarinier,Ph.D.5,6*, JoséeHébert,M.D.1,2,7,8 andGuySauvageau,M.D., Ph.D.1,6,7,81Hematology-OncologyDivision,Maisonneuve-RosemontHospital,Montréal,QC,Canada2TheLeucegene project atInstituteforResearchinImmunology andCancer,UniversitédeMontréal,Montréal,QC,Canada3DepartmentofComputerScienceandOperationsResearch,UniversitédeMontréal,Montréal,QC,Canada4InstituteforResearchinImmunology andCancer,UniversitédeMontréal,Montréal,QC,Canada5DepartmentofChemistry,UniversitédeMontréal,Montréal,QC,Canada6InstituteofResearchinImmunology andCancer– University ofMontreal,Montreal,QC, Canada7DepartmentofMedicine,FacultyofMedicine,UniversitédeMontréal,Montréal,QC,Canada8QuebecLeukemia Cell Bank,Montréal,QC,Canada

Monday,December5,2016:10:30AMPacificBallroom15-17(MarriottMarquisSanDiegoMarina)

89 TheRiskofMajorBleedingwithLow-Molecular-Weight-Heparins forVenousThromboembolisminDialysisPatients:TheQ-VTEStudy

AdiJ.Klil-Drori,MD1,2,JanieCoulombe,MSc3*,SharonJ.Nessim,MDMSc4,5* andVickyTagalakis,MD,MSc6,71DepartmentofOncology,McGillUniversity,Montreal,QC,Canada2JewishGeneralHospital,Montreal,QC,CAN3CenterforClinical Epidemiology,Jewish GeneralHospital,Montreal,QC,Canada4DivisionofNephrology,Department ofMedicine,Jewish GeneralHospital,Montreal,QC,Canada5DepartmentofMedicine,McGillUniversity,Montreal,QC,Canada6CenterforClinical Epidemiology,LadyDavisInstitute,Montreal,QC,Canada7DepartmentofMedicine,SirMortimerB.DavisJewish GeneralHospital,Montreal,QC,Canada

Saturday,December3,2016:10:30AMRoom31(SanDiegoConventionCenter)


AdiJ.Klil-Drori

• Background:Low-molecularweightheparins(LMWH)arenottraditionallyusedtotreatvenousthromboembolism(VTE)amongdialysispatientsbecausetheir renalclearance mayleadtolesspredictability inthedegreeofanticoagulationforagivendose.

• Theauthorsdetermined theriskofmajorbleedingwithLMWHcomparedwithvitaminKantagonist(VKA)useindialysispatientsdiagnosedwithVTEinarealworldsetting.


AdiJ.Klil-Drori

• Results:Inall,647dialysis patientswith VTEwere identified:467started VKA,82started LMWH,and96started both.Initiators ofLMWHwere 35dalteparin,26tinzaparin,19enoxaparin,and2nadroparin.

• Median (interquartile range,IQR)daily doseswere 12,500(7,500-17,570)IUdalteparin,16,080(13,540-20,000)IUtinzaparin,100(70-120)mgenoxaparin,and15,910(15,200-16,625)IUnadroparin.Median (IQR)durationofLMWHmonotherapy was 37(22-87)days,and132(65-235)forVKAmonotherapy.

• Morethan 90%ofLMWHmonotherapy was from 2004andonwards,and80%ofLMWHusershad cancer.

• Therewere 22majorbleeding events (86%gastrointestinal),20inVKAand2inLMWHusers.

• Nofatalbleeding occurred.

• Compared with VKAmonotherapy,LMWHmonotherapy was notassociated with majorbleeding(adjustedHR,1.21;95%CI:0.20-7.37).

527 TargetingPre-Leukemic StemCells inT-AcuteLymphoblastic Leukemia

BastienGerby,PhD1*,DiogoF.TVeiga,PhD1*,JanaKrosl,PhD1*,JulianneOuellette1*,AndréHaman1*,GenevièveLavoie,PhD1*,ImanFares,MSc1*,MathieuTremblay,Ph.D1*,VéroniqueLitalien1*,ElizabethOttoni1*,MilenaKosic1*,DominiqueGeoffrion1*,JoëlRyan1*,PaulMaddox,PhD2*,JalilaChagraoui,PhD1,AnneMarinier,Ph.D.1*,JoséeHébert,M.D.3,GuySauvageau,M.D.,Ph.D.1,BenjaminHKwok,PhD1*,PhilippePRoux,PhD1* andTrangHoang,PhD1

1InstituteofResearchinImmunology andCancer–University ofMontreal,Montreal,QC,Canada2UniversityofNorth CarolinaatChapelHill,ChapelHill,NC3TheLeucegene projectatInstituteforResearchinImmunology andCancer,UniversitédeMontréal,Montréal,QC,Canada

Sunday,December4,2016:5:30PMRoom10(SanDiegoConventionCenter)

527 Targeting Pre-Leukemic StemCells inT-AcuteLymphoblastic Leukemia

BastienGerby,PhD1

• CurrentchemotherapyofpediatricTcellacutelymphoblasticleukemia (T-ALL)efficientlyreducesthetumormasswith,however,undesirable longtermconsequencesandremainsineffectiveinadolescentandadultT-ALL.

• Furthermore,relapsecanbecausedbypre-leukemicstemcells (pre-LSCs)thatweresparedbycurrentprotocolsandevolvedtomalignancy.

• Adistinctivecharacteristic ofpre-LSCsistheircritical dependenceoninteractionswiththemicroenvironment forsurvival,whichguidedourstrategytotargetpre-LSCsusingniche-basedscreeningassays.


BastienGerby,PhD1

• Usingtransgenicmousemodelsthatcloselyreproducethehumandisease,theauthorshadshowedthattheSCL/TAL1andLMO1oncogenictranscriptionfactorsestablishapre-leukemicstatebyreprogrammingnormalpro-Tcellsintoaberrantlyself-renewingpre-LSCs(Gerbyetal.PloSGenetics,2014).

• Theynowprovidedirectevidencethatpre-LSCsaremuchlesschemosensitive thanleukemicblaststocurrentdrugs,duetoadistinctivelowerproliferativestateasassessedbyreal-timeimaginginacompetitiveassay.

• Theauthorsdesignedarobustprotocolforhigh-throughputscreening(HTS)ofcompoundstargetingprimarypre-LSCsthataremaintainedonstromalcellsengineeredforoptimalNOTCH1activationtomimickthethymic microenvironement.

• Screened1904compoundsandidentifiedUM0119979thatdisruptsbothcellautonomousandnon-cellautonomouspathways:UM0119979abrogatespre-LSCviabilityandself-renewalactivityinvivobyspecificallyinhibitingthetranslationofMYC,adownstreameffectorofNOTCH1,andpreventingSCL/TAL1activity.

• Incontrast:normalhematopoieticstem/progenitorcellsremainfunctional.

• Moreover,invivoadministrationofUM0119979efficientlyreducedtheleukemiapropagatingactivityofprimaryhumanT-ALLsamplesinxenograftedmice.

• Finally,inadditiontoSCL-LMO-inducedT-ALL,theseresultsrevealanovelpossibilityoftherapeuticinterventioninMYC-dependenthematologicmalignancies.


BastienGerby,PhD1

• Conclusion:

• Thisscreeningassay,builtonthegeneticdependenciesofpre-LSCs,revealedtheirvulnerabilitiestocompoundsthatinhibitboththeprimaryoncogenesandnon-cellautonomouspathwaystriggeredbythemicroenvironment.

• Theresultsillustratehowrecapitulatingtissue-likepropertiesofprimarycellsinhighthroughputscreeningisapromisingavenueforinnovationincancerchemotherapy.

75 Endosome-Mitochondria InterfaceControls IntracellularIronTrafficking inErythroidCells

AmelHamdi,PhD1,2,DanielGarcia-Santos,PhD1*,Tariq Roshan,MD3*,AlexSheftel,PhD4,5* andPremPonka,MD,PhD,FCMA1,21LadyDavisInstituteforMedical Research,Montreal,QC,Canada2DepartmentofPhysiologyandMedicine,McGillUniversity,Montreal,QC,Canada3McGillUniversity,Montreal,QC,Canada4SpartanBioscience Inc,Ottawa,Canada5HighImpactEditing,Ottawa,Canada

Saturday,December3,2016:10:00AMRoom3(SanDiegoConventionCenter)

75 Endosome-Mitochondria InterfaceControls Intracellular Iron Trafficking inErythroid Cells

AmelHamdi

• Inerythroidcells,morethan90%oftransferrin-derivedironentersmitochondriawhereferrochelatase insertsFe2+intoprotoporphyrin IX.However,thepathofironfromendosomestomitochondrialferrochelataseremainselusive.

• Theprevailingopinionisthat,afteritsexportfromendosomes,theredox-activemetalspreadsintothecytosolandmysteriouslyfindsitswayintomitochondriathroughpassivediffusion.

• Anopposingviewisthatthehighlyefficienttransportofirontowardferrochelatase inerythroidcellsrequiresadirectinteractionbetweentransferrin-endosomesandmitochondria(“kiss-and-run”hypothesis; PonkaBlood89:1,1997).

• Using3DliveconfocalimagingofreticulocytesfollowingtheirincubationwithMitoTrackerDeepRed(MTDR)andAlexaGreenTransferrin(AGTf),theauthorshavedemonstratedtransientendosome-mitochondriainteractions.

• Theyhavethuslyidentifiedapopulationofparticleslabeledwithbothfluorescentmarkers,representingendosomesinteractingwithmitochondria.FACSfollowedby2Dconfocalmicroscopyconfirmedtheassociationofbothorganellesinthedouble-labeledpopulation.


AmelHamdi

• Theauthorsexaminedwhetherreticulocytemitochondriainteractwithtransferrin(Tf)inacell-freesystem.LysatesofreticulocytespreviouslylabeledwithMTDRwereincubatedwithAGTf forvarioustimeintervals.

• IncreaseinthenumberofmitochondriaincontactwithfluorescentTf.Thiscanbepreventedbythepresenceofexcess,unlabeledFe2-Tf,butnotbyalbumin(Fig.1).Moreover,theadditionofunlabeledFe2-TftoreticulocytelysatesremovedAGTf frommitochondria,indicatingthatmitochondriafromreticulocytelysatesareassociatedwithTfR thatcanreversiblybindTf.


AmelHamdi

• Endosomescontainingmutatedrecombinantholotransferrin,whichcannotreleaseiron,remainassociatedwithmitochondria,whileendosomescontainingmutatedrecombinantapotransferrin,whichcannotbindiron,arenotassociatedwithmitochondria.Thesefindingsindicatethatendosomescontainingholo-Tfpromotetheirattachmentto,anddrivethedetachmentofapo-Tf-endosomesfrom,mitochondria,respectively.

• Byco-immunoprecipitationassay (frommurineeryhroleukemia [MEL]cellsandreticulocyteslysates), theauthorspurified thevoltage-dependentanionchannel2(VDAC2),whichislocatedattheoutermembraneofthemitochondrionwith DMT1.TheyconfirmedthecolocalizationofVDAC2andDMT1inMELcellsandreticulocytesbybothimmunofluorescenceandconfocalmicroscopy.Moreover,theyfoundasignificantdecrease inthenumberofmitochondriaincontactwithTf-endosomesafterdepletionof VDAC2inMELcellsoraftertreatmentofreticulocytelysateswiththemitochondrialuncoupler CCCP, furthersupportingtheconceptofaphysicalinteractionbetweenendosomesandmitochondria.

• DepletedMELcellsofVDAC2orinhibitedVADC2usingerastin (aspecificVDAC2inhibitorthataltersitsgating)andmeasured59Feincorporationfrom59Fe-Tfintoheme.Theyfounddecreased59FeincorporationintohemeofMELcellswithsilencedorinhibitedVDAC2supportstheideathatthisouter-membranemitochondrialproteinisinvolvedintheinteractionofendosomeswithmitochondria.

2306 Bortezomib Consolidation after Nonmyeloablative AllogeneicStemCell TransplantationLeadstoaHighIncidenceofImmunophenotypic CompleteResponse inYoungand/orHigh-Risk MultipleMyeloma Patients

RichardLeBlanc,MD1,ImranAhmad,MD2,Rafik Terra,PhD3*,SéverineLandais,PhD2*,MichaelSebag,MD,PhD4,Emilie Lemieux-Blanchard,MD5,NadiaM.Bambace,MD2,LeaBernard,MD2,SandraCohen,MD1,Jean-SebastienDelisle,MD,PhD6,ThomasKiss,MD2,SilvyLachance,MD6,Denis-ClaudeRoy,MD2,GuySauvageau,MD,PhD7 andJeanRoy,MD81ServiceofHematologyandMedical Oncology,Department ofMedecine,Maisonneuve-RosemontHospital,Montreal,QC,Canada2DivisionofHematologyandMedical Oncology,StemCell TransplantProgram,Department ofMedicine,University ofMontreal,Maisonneuve-RosemontHospitalCIUSSSEast,Montreal,QC,Canada3ImmunologyLaboratory,Department ofHematologyLaboratory,Maisonneuve-RosemontHospital,Montreal,QC,Canada4McGillUniversityHealthCentre,Montreal,QC,Canada5Hemato-oncologydepartment,CHUM,Montreal,QC,Canada6DivisionofHematologyandMedical Oncology,StemCell TransplantProgram,Department ofMedicine,University ofMontreal,Maisonneuve-RosemontHospitalCIUSSSEastMontreal,Montreal,QC,Canada7DepartmentofMedicine,IRIC/University ofMontreal,Montreal,QC,Canada8DivisionofHematologyandMedical Oncology,University ofMontreal,Maisonneuve-RosemontHospitalCIUSSSEastMontreal,Montreal,QC,Canada

Saturday,December3,2016,5:30PM-7:30PMHallGH(SanDiegoConventionCenter)

2306 BortezomibConsolidationafter NonmyeloablativeAllogeneicStemCell TransplantationLeadstoaHighIncidenceofImmunophenotypic CompleteResponse inYoungand/orHigh-RiskMultipleMyeloma Patients

RichardLeBlanc

• Allogeneic stemcelltransplantation(alloSCT)istheonlycurativemodalityfornewlydiagnosedmultiplemyeloma(NDMM)patients(pts).

• Theauthorshavepreviouslyshowninalargecohortof92ptsthatrelapseremainscommon(49%)andtheincidence/severityofchronicGVHDissignificant(79%)aftertandemauto-alloSCTinNDMMpts(Ahmadetal.BMT2016;51:529).

• Theyhypothesizedthatatandemauto-nonmyeloablative (NMA)alloSCT followedbybortezomib (btz)consolidationmightbesafe,whiledecreasingboththeseverity/incidenceofchronicGVHDandtheriskofrelapseinyoungand/orhigh-riskNDMMpts.

• Inaddition,theyhypothesizedthatbortezomibmightfurtherincreasedepthofresponsesafteralloSCT.


RichardLeBlanc

• Methods:NDMMptswitheitherISSstageIII,plasmacellleukemia,abnormalcytogeneticsdefinedast(4;14)withISSIIorIII,t(14;16),t(14;20),17p-,1p-,or1q+in≥10%ofpurifiedplasmacellsorage≤50yearswitha6/6siblingor8/8unrelateddonorwereprospectivelyenrolledinthisphaseIItrial.

• Afterabtz-basedinductionwith≥partialresponseandautologous(A)SCT,outpatientNMAalloSCTwasperformedwitheitheraconditioningoffludarabine30mg/m2 x5daysandcyclophosphamide300mg/m2 x5days(siblingdonor)orfludarabine30mg/m2 x3daysandTBI2Gy(unrelateddonor),followedbyG-CSFmobilizedstemcellsinfusion.

• AcuteGVHDprophylaxisconsistedoftacrolimusandmycophenolatemofetil. Btz 1.3mg/m2 SCevery2weekswasstartedonday+120afteralloSCT for1year.

• BonemarrowaspiratesbeforealloSCT,beforestartingbtz andevery3monthsthereafterwereprospectivelycollectedfor2yearsinordertoassesstheimpactofbtz onminimalresidualdisease(MRD)byahighlysensitive(≥10-5)multiparametric flowcytometryusingthe8-colorEuroflowprotocolevaluating≥10x106cells/specimen.

• MRDnegativitywasdefinedasthedetectionof < 30clonalaberrantplasmacells.ResponseevaluationisbasedonIMWGcriteriaincludingimmunophenotypic completeresponse(iCR)definedasastringentCR(sCR)plusanegativeMRD.Immunophenotypic remission(iR)isdefinedasMRDnegativityregardlessofotherdiseasestatus.


RichardLeBlanc

4677 TandemAutologousFollowedByNonmyeloablative AllogeneicTransplantationinRelapsedHighRisk Follicular Lymphoma LeadstoExcellentLongTerm Progression-FreeSurvival after 8Years ofFollow-upClinical AllogeneicTransplantation:ResultsPosterAbstractsSession: 732.Clinical AllogeneicTransplantation:Results:PosterIII

Monday,December 5,2016,6:00PM-8:00PMHallGH(SanDiegoConventionCenter)

MagalieTardif,MSc,MDStudent1*,ImranAhmad,MD2,NadiaM.Bambace,MD2,LeaBernard,MD2,LambertBusque,MD2,Jean-SebastienDelisle,MD,PhD3,ThomasKiss,MD2,IsabelleFleury,MD2,SilvyLachance,MD3,LuiginaMollica,MD,PhD4,CélineNkoué,MD2*,Denis-ClaudeRoy,MD2,JeanRoy,MD2 andSandraCohen,MD51UniversityofMontreal,Maisonneuve-RosemontHospitalCIUSSSEast,Montreal,QC,Canada2DivisionofHematologyandMedical Oncology,StemCell TransplantProgram,Department ofMedicine,University ofMontreal,Maisonneuve-RosemontHospitalCIUSSSEast,Montreal,QC,Canada3DivisionofHematologyandMedical Oncology,StemCell TransplantProgram,Department ofMedicine,University ofMontreal,Maisonneuve-RosemontHospitalCIUSSSEastMontreal,Montreal,QC,Canada4HôpitalMaisonneuve-Rosemont,Montreal,QC,Canada5ServiceofHematologyandMedical Oncology,Department ofMedecine,Maisonneuve-RosemontHospital,Montreal,QC,Canada

4677 TandemAutologousFollowedByNonmyeloablative AllogeneicTransplantationinRelapsedHighRisk FollicularLymphoma LeadstoExcellentLongTerm Progression-FreeSurvival after 8Years ofFollow-up

MagalieTardif,MSc,MDStudent

Prospective protocol initiated in April 2003for pts with high risk relapsed FL as defined by chemorefractory disease, early 1st relapse, >1st relapse or transformation into aggressive histology.

At least one therapy was attempted to document chemosensitivity prior to ASCT.

Rgardless of disease status prior to transplant, pts underwent ASCT followed 3 months later by an outpatient NMT from an HLA-identical sibling.

NMT comprised 5 days of fludarabine 30 mg/m2/day and cyclophosphamide 300mg/m2/day followed by an infusion of >2x106CD34+ cells/kg.

GVHD prophylaxis: tacrolimus starting on day (D) -8 to achieve levels of 8-12 nmol/L then tapered off by D+100 or D+180 depending on disease risk and of

Report on 40 pts with a median f/u of 8 yrs.



• UpuntilJuly2015,40ptswereenrolledwithamedianageof50 yrs (34-65).

• Ptshadpreviouslybeentreatedwithamedianof3linesoftherapy(2-6).

• MediantimefromdiagnosistoASCTwas33months.DiseasestatusatASCTwas:14CR,16PRand10refractory.

• ConditioningforASCTincludedBEAM/BEAC(n=39),andCy-TBI(n=1).

• 4ptsreceivedradiotherapyafterASCTtositesofpreviouslybulkydisease.

• MediantimebetweenASCTandNMTwas138days(75-238).

• PreNMTdiseasestatuswas:25CR,12PRand3refractory.

• EngraftmentwaspromptinallptsafterASCTandmedianneutrophilandplateletrecoverywererespectively13days(0-19)and0day(0-18)postNMT.

1535 Modeling ofPediatric AcuteMegakaryoblastic Leukemia Using CordBloodStem/Progenitor CellsOncogenes andTumor SuppressorsProgram: PosterAbstractsSession: 603.Oncogenes andTumor Suppressors:PosterI

Saturday,December 3,2016,5:30PM-7:30PMHallGH(SanDiegoConventionCenter)SophieCardin,PhD1*,LouiseLaramee2*,TaraMacRae,M.Sc.3*,Jalila Chagraoui,PhD4,GuySauvageau,MD,PhD5,R.KeithHumphries,MD,PhD6,JoséeHébert,M.D.7,BrianT.Wilhelm,PhD8* andSoniaCellot,MD,PhD91Medecine,Universite deMontreal,Montreal,QC,Canada2CHUSJ,Montreal,Canada3InstituteforResearchinImmunology andCancer,Montréal,QC,Canada4LaboratoriesofMolecular Genetics ofHematopoietic StemCells,InstituteforResearchinImmunology andCancer(IRIC),Montreal,QC,Canada5DepartmentofMedicine,IRIC/University ofMontreal,Montreal,QC,Canada6TerryFoxLaboratory,BritishColumbiaCancerAgency,Vancouver,BC,Canada7TheLeucegene project atInstituteforResearchinImmunology andCancer,UniversitédeMontréal,Montréal,QC,Canada8IRIC/University ofMontreal,Montreal,QC,Canada9UniversityofMontreal,Montreal,QC,CAN

1535 Modeling ofPediatric AcuteMegakaryoblastic LeukemiaUsing CordBloodStem/Progenitor Cells

SophieCardin,PhD1*,

• Pediatric acutemegakaryoblasticleukemia (AMKL)accounts for10%ofchildhoodacutemyeloid leukemia (AML)casesandremainsahighfatalitycancer.CBFA2T3-GLIS2,NUP98-KDM5A,RBM15-MKL1andMLLgene rearrangementsarerecurrentaberrationsthat aremutuallyexclusiveandfound atsimilar frequencies inhalf thecasesofpediatric AMKL.

• Therecently identifiedCBFA2T3-GLIS2andNUP98-KDM5Achimeric oncogenesareassociatedwith inferior outcomes (overall survival,OS:~30%)comparedtopatientsharboring theRBM15-MKL1gene fusion(OS~70%).

• ToinvestigateNUP98-KDM5Adriven leukemogenesis,humancell lines andmousemodels wereengineeredusing overexpressionofthechimeric oncogene inCD34+cordblood (CB)stem/progenitor cells.

1535 Modeling ofPediatric AcuteMegakaryoblastic LeukemiaUsing CordBloodStem/Progenitor Cells

SophieCardin,PhD1*,

• cDNAoftheNUP98-KDM5Afusion:nuclearporeproteinnucleoporin98(NUP98)genefusedtothehistonelysinedemethylase5A(KDM5A)gene,wasclonedintoaMNDUlentiviralvectorcarryingaGFPreportergene.

• Usingoptimizedcultureconditions,10,000freshlyisolatedCB-CD34+ (day0)cellswereseededinmultiplewells invitroandtransducedwitheitherNUP98-KDM5Aorcontrol(CTL)vectors.

• Xenotransplantationof75%ofday7cellsinimmunodeficient miceresultedinthedevelopmentofovertAMKLin1of3miceafter32weeks.Recipientmouseboneswerewhiteandbrittle,andthemarrowcavityinfiltratedby30%hCD45loCD61+GFP+ leukemicblasts,withtypicalmegakaryoblasticmorphology.

• Theleukemicblastswerealsodetectedinblood(5%),andinenlargedspleen(0.2%).SecondarytransplantationofisolatedAMKLcells(frombonemarrowandspleen)wasperformed,alongwithexpressionprofilingbyRNAsequencing.

2372 BurdenofRelapseFollowing AllogeneicHematopoietic StemCell TransplantationonHealthCareResourceUtilization intheManagementofAcuteLeukemiaandMyelodysplastic SyndromeHealthServicesResearch—Malignant ConditionsProgram:OralandPosterAbstractsSession: 902.HealthServicesResearch—MalignantConditions:PosterI

Saturday,December 3,2016,5:30PM-7:30PMHallGH(SanDiegoConventionCenter)SilvyLachance,MD1,Joelle Bibeau2*andJeanLachaine3*1DivisionofHematologyandMedicalOncology,StemCell TransplantProgram,Department ofMedicine,University ofMontreal,Maisonneuve-RosemontHospitalCIUSSSEastMontreal,Montreal,QC,Canada2PeripharmInc,Montreal,QC,Canada3FacultyofPhamacy,Universite deMontréal,Montreal,QC,Canada

2372 Burden ofRelapseFollowing AllogeneicHematopoietic StemCell TransplantationonHealthCareResourceUtilization intheManagementofAcuteLeukemia andMyelodysplastic Syndrome

SilvyLachance

1226 DonorLymphocytesDepletedofAlloreactiveT-Cells(ATIR101) ImproveEvent-FreeSurvival(GRFS)andOverallSurvivalinaT-CellDepletedHaploidenticalHSCT:Phase2TrialinPatientswithAMLandALL

Denis-ClaudeRoy,MD1,SilvyLachance,MD2,JeanRoy,MD3,IrwinWalker,MBBS4,JohanMaertens,MD,PhD5*,Jean-SebastienDelisle,MD,PhD2,StephenRonanFoley,MD6,PhilippeLewalle,MD-PhD7*,EduardoOlavarria8*,DominikSelleslag,MD9,ManfredRüdiger,PhD10*,JurjenVelthuis,PhD10*,LisyaGerez10*,JeroenRovers,MDPhD10*,HalvardBonig,MD,MA11 andStephanMielke,MD121BloodandMarrow TransplantationProgram/DivisionofHematology&Oncology,HopitalMaisonneuve-Rosemont/Universite deMontreal,CIUSSSEastMontreal,Montreal,QC,Canada2DivisionofHematologyandMedicalOncology,StemCell TransplantProgram,Department ofMedicine,University ofMontreal,Maisonneuve-RosemontHospitalCIUSSSEastMontreal,Montreal,QC,Canada3DivisionofHematologyandMedicalOncology,University ofMontreal,Maisonneuve-RosemontHospitalCIUSSSEastMontreal,Montreal,QC,Canada4JuravinskiHospitalandCancerCentre,McMasterUniversity,Hamilton,ON,Canada5DepartmentofHematology,University HospitalGasthuisberg,Dept.ofHematology,Leuven,Belgium6McMasterUniversity,Department ofMedicine,Juravinski HospitalandCancerCentre,Hamilton,ON,Canada7LaboratoryofExperimentalHematology,JulesBordetInstitut,Bruxelles,BEL8CentreforHaematology,ImperialCollege LondonatHammersmithHospital,London,UnitedKingdom9AZSt-Jan BruggeAV,Brugge,Belgium10KiadisPharma,Amsterdam-Duivendrecht,Netherlands11GermanRed CrossBloodCentreandInstituteforTransfusionMedicine andImmunohematology,Johann-Wolfgang-GoetheUniversity,Hematopoietic Cell ResearchGroup,Frankfurt,Germany12DivisionofHematologyandOncology,Department ofInternalMedicine II,WürzburgUniversity Medical Center,Würzburg,Germany

Monday,December5,2016:6:30PMRoom30(SanDiegoConventionCenter)

ChallengeinHaploidentical DonorTransplantation

HAPLOIDENTICALDONOR

LEUKEMIA PATIENT

XX

XX X

EffectsATIR101procedure• SelectiveremovalofGVHD-causingT-cells

• Preservationoftheimmunerepertoire

• Keyimmunecellsareretainedtoprotectagainstinfections

• T-cellsdirectedagainstleukaemic antigensareretained

• ATIR101manufacturing

Potentialbenefits

• 5daymanufacturingprocess

• Release data

• PerformedinadvanceofHSCT

procedure

• Cells cryopreserveduntil

infusion

1. Immune cells collected and mixed

ex vivo

2. Activation of donor T- cells

GVHD causing T-cells donor cells are activated by patient cells

Patie

ntDo

nor

3. TH9402 addition

Proprietary photosensitizing reagent TH9402 is added.

TH9402 is retained only in activated donor T-cells

4. Exposure to light

Cells are exposed to light, which activates TH9402, generating toxic oxygen radicals

5. ATIR infusion

Cells are formulated for infusion

Infusion done 28-42 days after Haplo HSCT

Day 1 - 4 Day 5

• Phase IIclinical trial• Aimistodevelopanimmunosuppressant-free transplantregimenforhaploidentical donor

transplantation

• BasedonT-celldepletion:CD34+-megadoseapproach (Perugia)

• Pre-emptivepost-HSCTadministrationofdonor lymphocytes(ATIR101)depleted ofalloreactiveT-cells

• AvoidGVHD• Reduceinfections/TRM/relapse

Myeloablativeconditioning

T-cell depleted stem cell graft from family member

Isolation

No prophylactic immunosuppression

CD34+ PBMC graft

Donor lymphocyte infusion

28 – 32 days post-HSCT

Haplo HSCT ATIR101 infusionConditioning

Selective photodepletion to eliminate GVHD-causing T-cells

Donordonor

• 1 Mrozek K, et al. JCO 2012, 30 (36):4515-4523 • 2Armand P, et al. Blood 2014, 123 (23); 3664-3671

• Patient&DonorcharacteristicsDiagnosisPatient & Donor

• N=23patients(HSCT+ATIR101)

• Medianpatientage(range):41years(21–64)

• Gender:13female,10male

• Mediandonorage(range):33years(21- 61)

• HLAmatching(HLA-A,B,DR)

– 3/6match:16

– 4/6match:6 5/6match:1(7/10match)

• Donors:

– Father/mother 4(17%)

– Sibling 9(39%)

– Son/daughter 9(39%)– Other 1(4%)

• Acutemyeloidleukemia–N=16(70%)

– 11inCR1

– 5inCR2

• Acutelymphoblasticleukemia –N=7(30%)

– 4inCR1

– 3inCR2

• Cytogeneticriskprofile1:

– Favorable 0– Intermediate

9(39%)

– Adverse 14(61%)

• Disease-riskindex2:

– Lowriskindex0

– Intermediateriskindex 10(43%)

– Highriskindex13(57%)

• Transplantationcharacteristics(n=23patients)Conditioning

HSCT

HSCT

Prophylaxis GVHD

• TBI(1200cGy;n=11)ormelphalan(120mg/m2;n=12)

• Thiotepa (10mg/kg),fludarabine (30mg/m2x 5d)andATG(2.5mg/kgx 4d)

• CliniMACS®CD34isolationsystem(Miltenyi Biotec)

• Target:8-11x106 CD34+cells/kg,withmax.of3x104

CD3+cells/kg

• NoGVHDprophylaxis

• CMV/EBVmonitoring

• Prophylacticuseofganciclovir /foscarnet (CMV+recipient/donor)

• NoGradeIII-IVGVHD

• GradeIIacuteGVHDin3patients

• ChronicGVHD:1patient

Graft Median(cells/kg)

range

CD34+ 10.9x 106 3.2– 24.4x106

CD3+ 0.28x 104 0– 1.8x 104

Engraftment Median(days) range

Neutrophils 12 8– 34

Platelets 12 9– 35

ASH2016,Session711,OralpresentationonMondayDecember 5,18h30

1226DonorLymphocytesDepletedofAlloreactiveT-Cells(ATIR101)ImproveEvent-FreeSurvival(GRFS)andOverallSurvivalinaT-CellDepletedHaploidenticalHSCT:Phase2Trial inPatientswithAMLandALLDenis-ClaudeRoy

ISCT 2018 GENERAL MEETING

Cell & Gene Therapy RevolutionMontreal: March 9-10, 2017

Invitationtomeetings:

• Québec City: 27 janvier 2017• Montréal: 7 avril 2017

Québec Highlights › wp-content › uploads › 2019 › 12 › Quebec.pdf• CLEC-2, the PDPN receptor, is expressed on normal platelets and was found to be necessary for the separation

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