Bridging the Gap Between Preclinical/Translational Data to Clinical Applications - Eric raymond - Tumour Models London 2016

BridgingtheGapBetweenPreclinical/TranslationalDatato

ClinicalApplicationsEricRaymondMD,PhD

ChairofMedicalOncology@Groupe Hospitalier ParisSaint-Joseph- France

[email protected]

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


S49076 (MET/AXLi) delays tumor progression and angiogenesis in a transgenic HCC mouse model

Livervolume Bloodflowvelocity

S49076delaystumorprogression S49076inhibitsproliferationandnodulesize

CD31stainingandquantificationat12Wand16W

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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



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



11027 ♂, 63 years SCC maxilla




7067 ♀, 50 years SCC oropharynx

7872 ♀, 66 years SCC oropharynx

Control Debio 1143 CisplatinD1143

CisplatinCarboplatinD1143

CarboplatinHE

Caspase 3

Ki67

Pt14859

Effectsofdrugsinexvivoexposedsquamouscellcarcinomaofthehead&neck


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

[email protected]

Health & Medicine

Bridging the Gap Between Preclinical/Translational Data to Clinical Applications - Eric raymond - Tumour Models London 2016