bridging the gap between preclinical/translational data to clinical applications - eric raymond -...
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BridgingtheGapBetweenPreclinical/TranslationalDatato
ClinicalApplicationsEricRaymondMD,PhD
ChairofMedicalOncology@Groupe Hospitalier ParisSaint-Joseph- France
Disclosures• Pfizer• Novartis• EliLilly• Ipsen• Oncoethyx• Genoscience Pharma
Theevolvingclinicaltrialsparadigmforthedevelopmentofmolecularly-targetedtherapeuticagents
TheeasygoingdrugdevelopmentpatternOneoncogenicdriver– Onemodel– Oneselectionbiomarker
Bcr-ablKITEGFRROSALKRAF/RASEtc…
Oncogenicdriver(mutation/translocation)
- Mutatedcelllines
- Transgenicmousemodels
- SelectedPDXcarryingthemutation
Patientselectionbasedontheexpressionofthedrivingmutationontumorbiopsyatstudyentry
Preclinicalmodels Clinicaltrials
- Preclinicalmodelsarenotalwaysusefulwhentheclinicaldevelopmentisobvious- Unfortunately,whileusuallyallowingfasttrackdrugapproval,thispatterndoesnotconcernmosttumors
Newparadigms
• Co-clinicaltrialscombiningstudiesdoneinpatientsandinamousehospital
• Usingxenograftstotestbiologicalparametersthatarebarelyaccessibleduringclinicaltrials
• UsingPDXtotrytomatchpatientsoutcomewiththatinanimalmodels
• Usinggeneticallyengineeredmousemodelsthatrecapitulatemaingeneticaberrations
observedinhumantumors
• Ex-vivomonitoringofdrug-inducedbiologicalchangesintumorsharvestedfrom
patientspriorandduringtrials
Co-clinicaltrialsinhumanandmousehospitals
- Engraftmentratesmaydownsizethenumberof
patientsbenefitingofthisapproach
- DosingandPK/PDparametersmaygreatlydiffer
frommouseandhumanforaparticulardrug
- Antibodiesinhumanandmousearestrikingly
different
- Tumorgrowthandresponsetotreatmentmaybe
inconsistentwiththedurationoftreatmentin
human
- Currently,novalidatedexperiencesupportsthe
increasingcostofrunningmouseandhumantrials
inparallel
PDX&GEMM
Thebasis Thepitfalls
Cellular&MolecularComponentsofCarcinomaMicroenvironmentsareoftenverycomplexandchanging
EndothelialcellsPericytesVEGFR-PDGFR
Tcells(CD4-Treg)CD4:PD1-CTLA4-CD28Treg:CD73-CD39
DendriticcellsPDL1-PD1-MSHII-CD80/86
TumorassociatedmacrophagesCXCR4-TGFβR
TumorcellsTGFβR-MET-PDL1
TGFβHGFFGF19IL8IL10
SDF1/CXCL12
FibroblastsFGFR
Doweneedpreclinicaldataandwhatfor?
• Modelsareusedtoprovideinsightswhendesigningclinicaltrials:
• Provideevidencesofantitumoractivityinvarioustumortypes
• Suggestmarkers tobetterselectpatients/tumorssensitivityorresistanttodrugs
• Identifysafety,pharmacokinetic,andschedulingissues toprepareforIND
• Initialdosing
• Doseescalationscheme
• Scheduleforoptimaldoseexposureandminimaltoxicity
• Identifyadverseeventsofpotentialspecialinterestaccordingtothemechanismofaction
Pitfallscommonlyobservedusingmodels
• Modelsremainoftenincapableofpredictingoutcomeinclinicaltrials• Nonealltumortypesaregeneratingpreclinicalmodelsthatarerelevantwithclinicaloutcomes(neuroendocrinetumors,pancreaticadenocarcinoma,hepatocellularcarcinoma,etc…)
• Celllines,spheroids,xenografts,PDX,andtransgenicmiceencompassonlypartlythevariouscomponentsofhumantumorheterogeneity
• Experimentsandmodelshavelimitedtimeframesandcannotrecapitulateepigeneticconstraintsofnaturalhistoryofdiseasesandtherelatedbiologicaldriftsassociatedwithtumorprogressionandmultipletreatmentsusedinhuman
MoredemandingdrugdevelopmentpatternsMultiple/unknownoncogenicdriver– Unclearmodel–unknownselectionbiomarker
Variouscomponentsofthetumormicroenvironment- Angiogenesis- Local
immunosuppression- EMT- Metabolism
Epigeneticdrivers(expression)
? Patientselection?
Preclinicalmodels Clinicaltrials
- Severalnovelanticanceragents(antiangiogenics,checkpointinhibitors,microenvironmentsignalinginhibitorssuchasinhibitorsofMET,CXCR4,TGF-beta,etc…)arevariablyexpressedintumorsandtheirexpressionarecommonlymodulatedthroughvariousepigeneticchanges
- Preclinicalmodelsareusuallycomplextodevelopbutarehighlyimportanttoprepareclinnical trials
Threeusefultypesofmodelstoevaluatedrugactivity
Models Aimsofstudies cellularcomponents Example
HumancancercellXenografts
Testspecifichypothesis
Humancancercells+mouse
microenvironment
Antiangiogenic(VEGFRinhibitors)
Transgenicmodels
Recapitulatecarcinogenicconditions
Mousecancercell+mouse
microenvironment
Microenvironmentsignalinginhibitors(TGF-betareceptor
inhibitors)
Exvivotumorexposures
Evaluateeffectsoncancercellsintheir
innatestroma
Humancancercellsinhuman
microenvironment
Microenvironmentsignalinginhibitors+immunecheckpoint
inhibitors
XenograftmodelsExampleusingrenalcellcarcinomamodelstoevaluateantiangiogenicdruginducedchangesintumors
HumancancercellXenografts
Human cancercells
Mouse stromacells
Clinicaltrialinmice CAKI-1and786-0Oncotarget 2016
CAKI-1&786-0tumorsaresensitivetosunitinib,progressionoccurringbetween40-60daysofexposure
Sunitinib
Sunitinib
CAKI-1
786-0
Oncotarget 2016
GeneexpressionprofilingoftumorsatthetimesofresponseandprogressionshowedtrendsformesenchymaldifferentiationinRCCxenografts
• Sunitinibtreatmentaffectedgenesandproteinsinvolvedingrowthanddifferentiationinfavorofamoreepithelialphenotypethanuntreatedtumors
• Tumorsthatstarttoprogressinsunitinibtreatedmiceexpressedgenesinvolvedinangiogenesisanddifferentiation,resulting1. Inanexacerbatedmesenchymal
phenotype2. Intheexpressionofendothelialgenes
CAKI-1 786-0
Oncotarget 2016
Renalcancercellsattheperipheryofvesselsexpressmarkersofvasculardifferentiationconsistentwithvascularmimicry
CAKI-1786-0
Oncotarget 2016
Atthetimeofprogressionundersunitinibtreatment,renalcancercellsre-expressvimentin,CD133andotherproteinssuchasCXCR4,AKT,andmTOR*
*ResultsdisplayedforCAKI-1withsimilarpatternsfor786-0
mRNA byRT-PCROncotarget 2016
Secondlinetherapywitheverolimusintumorsthatprogressedfirst-linesunitinibdelaystumorgrowthandaffectstumorangiogenesis
Oncotarget 2016
Second-linetherapywitheverolimusinducesstrongE-cadherinexpressionandreducesvascularmimicry
Oncotarget 2016
TransgenicmodelsExampleusingtheASVBmodeltoevaluatedrugsactingonthetumormicroenvironmentsignaling
Transgenicmodels
Mouse cancercells
Mouse stromacells
Transgenic mouse model developing hepatocellular carcinoma(ASV-B)
Nodular / 12W Diffuse / 16WDysplasia / 8W
CD31
x20x20 x20
Transgene on the Y chromosome
Fusion between the human anti-thrombin promotor (expression only in liver cells) and the large T oncogene of SV40
* x20
*x20x20
HES
CourtesyAnnemilaï Tijeras-Raballand – AFROncology
Galunisertib delays tumor progression and angiogenesis in a transgenic HCC mouse model
Livervolume Bloodflowvelocity
At12weeksAt16weeks
CD31stainingandquantificationat16W
CourtesyAnnemilaï Tijeras-Raballand – AFROncology
S49076 (MET/AXLi) delays tumor progression and angiogenesis in a transgenic HCC mouse model
Livervolume Bloodflowvelocity
S49076delaystumorprogression S49076inhibitsproliferationandnodulesize
CD31stainingandquantificationat12Wand16W
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CourtesyAnnemilaï Tijeras-Raballand – AFROncology
ExvivotestingofmacroscopichumantumorspecimensExampleofusingtheASVBmodeltoevaluatedrugsactingonthetumormicroenvironmentsignaling
Exvivotumorexposures
Human cancercells
Human stromacells
Drug selection Drug testing Data analysis
patient Doctor
Therapeutic proposal
BiopsyBloodtests Daysfrombiopsy
15 days
EricRAYMOND – Kinasesoncogéniques etthérapies ciblées – Académie dessciences– 23avril 2013
Strategiestodevelopprecisionmedicineandpersonalizedtherapeuticsinroutinepractice
Drugtestingplatformintheclinic
O2
CO2
O2/CO2/T°monitor
48h-96h
Surgical specimenBiopsy
Tissueslicing
Control
DrugA
DrugB
Treatment & culture
CourtesyAnnemilaï Tijeras-Raballand – AFROncology
CourtesyAnnemilaï Tijeras-Raballand – AFROncology
PROCESSFROMSURGICALSPECIMEN
DeGraafIAMetal.,NatProtocols.2010
ü TumorcoresarepreparedusingasurgicalpunchandtransferredintocoreholderoftheTissueSlicer
ü Slicesof8mmareperformedandtransferredto6-wellplatesusingaspatulatoavoidtissuedamages
Control
Biopsyunderlocalanesthesia
Freshtumor-tissueculturedfor48hTotesttargetedtherapies
Biomarkersanalysisbyimmunofluorescence
ExVivoevaluationofdrugsinindividualpatients:precisionmedicine
DrugA
DrugB
EricRAYMOND – Kinasesoncogéniquesetthérapiesciblées– Académiedessciences– 23avril2013
InteligenceMedicineProteinandnucleicacidexpressions
Chemo-biogramEffectsofdrugsontissuebiomarkers
ID Patient Cancer type
8590 ♂, 56 years SCC larynx
1148 ♂, 60 years SCC hypopharynx
6508 ♂, 60 years SCC oropharynx
6934 ♂, 60 years SCC larynx
10635 ♂, 60 years SCC larynx
11027 ♂, 63 years SCC maxilla
0330 ♂, 61 years SCC oropharynx
14951 ♂, 55 years SCC oropharynx
14859 ♂, 57 years SCC oropharynx
7067 ♀, 50 years SCC oropharynx
7872 ♀, 66 years SCC oropharynx
Control Debio 1143 CisplatinD1143
CisplatinCarboplatinD1143
CarboplatinHE
Caspase 3
Ki67
Pt14859
Effectsofdrugsinexvivoexposedsquamouscellcarcinomaofthehead&neck
CourtesyAnnemilaï Tijeras-Raballand – AFROncology
OTX-008 relocates galectin-1 in the nucleus of cancer cells and reduces cellular proliferation in head & neck squamous cell carcinoma tumors ex vivo (IHC)
NCI-AACR-EORTCmeeting2012
CAPRA:Clinicaltrialdesign
- Everolimus(RD)- CarboplatinAUC2- Paclitaxel60mg/m²
Chemoradiationtherapy
PhaseIdoseescalation:30-50mg/weekeverolimuscombinedwithAUC2carboplatinand60mg/m2paclitaxelweekly
PhaseIIevaluation(twostageSimondesign)
Weeklyx 9 cycles
CAPRA
FaivreSetal.
Caspase3
P-S6k
Ex-vivo ActivityofeverolimusinFreshHNSCCTumors
Ki67
CTR48h Everolimus(0,1µMfor48h)
FaivreSetal.
Ki67
p-S6K
Baseline Post-CAPRA
IHC
IHC
IF
Pre- &Post-TreatmentBiomarkersinBiopsies
FaivreSetal.
WaterfallplotevaluationofpatientstreatedwithCAPRA(RECIST1.1)
FaivreSetal.
Conclusions• Modelsoftenusedtopredictdrugactivityinpatients,maybemoreaccuratetounderstandspecificchangesintumorsanddevelopassumptionsforfurtherclinicaltesting
• Developingmodelsthatrecapitulatethecomplexityofhumantumormicroenvironmentswillbemoreandmoredesirableasdrugtargetingthelocalimmunosuppressionandthemicroenvironmentsignalingwilldevelopintheclinic
• Usingexvivodrugtestinginfreshhumantumorspecimensappearsfeasibleinclinicaltrials
http://pamm2017.org
Presentyourwork!
ThanksforyourattentionEricRaymondMD,PhD
ChairofMedicalOncology@Groupe Hospitalier ParisSaint-Joseph- France