MBP1018, Feb 1, 2012

Novel models and methods for assessment of new targeted agents in oncology
DavidHedley MedicalOncologyandHematology DeptMedicalBiophysics

PancreaticCancer: TheClinicalProblem
65 yrs male, presenting with back pain Unresectable due to major blood vessel encroachment Commenced chemo/radiation March 2006
Feb2006 June2006

Gemcitabine erlotinib (NCICCTGPA3)

PrimaryPancreaticCancerXenografts
>85%ofpancreatectomy samplesgrow Morphologysimilartoprimarycancerwhen implantedatorthotopic site:
1. Nearclinical testingofnovelagents 2. InternationalCancerGenomeConsortiumproject 3. Translationalresearchintounderlyingbiology

Simulating clinical trials using primary pancreas cancer xenografts, grown orthotopically
Heart

Liver

Simulating clinical trials using primary pancreas cancer xenografts, grown orthotopically
Heart

Liver

AktSer473

AberrantactivationofKras dependent pathwaysinpancreaticcancer

Wenowhavedrugstoblocksignalingpathways wheretheupstreameffectorsconverge buthowsafearethey?

NVPBEZ235

Clinical trialtesting BEZ235 in 5 pancreas cancer patients as orthotopic implants in mice

H&E pAkt

OCIP16

Effects of daily dosing with BEZ235 on tumour weights

OCIP17

OCIP18

OCIP19

OCIP21

Howfarcanyoupushchronicdosing withaPI3kinaseinhibitor?
Followingacutesingledose, pAkt goesdown,but recoversat24hr Inchronicdosing,westill findpAkt attheendof treatment Miceloseweightathigher doselevels

Youcannothave100% inhibitionofamajor signalingpathway100% ofthetime! (Hoeflich paper)

Wheredowegonow?
Genomics individualizedmedicine rationalselectionofdrugcombinations determinedbyasyettobedeveloped analyticalplatforms Targetthetumour microenvironment geneticallylesscomplex angiogenesis(butsofarabust) tumour stroma tumour hypoxia

Similartopancreascancerpatients,orthotopic primaryxenograftscontaindensefibrousstroma, enrichedinmesenchymal cells

MassonTrichrome SmoothMuscleActin

Yauchetal.Nature2008

Hedgehogpathway

HedgehoginhibitorIPI926attenuatesfibrous stromainKPCmousemodelofpancreaticcancer Oliveetal.,Science2009;324:145761

Controlgemcitabine

IPI926gem+IPI926

H&E

CollagenI

Effects of anti-SHH antibody 5E1 given by weekly i.p. injection to mice bearing orthotopic OCIP19 xenografts

Control-mouse 1

5E1-mouse 1

-smooth muscle actin

200m

Hedgehog gene expression in OCIP tumors with 5E1 treatment

Pooled tumors: control n=4; 5E1 n=3 Values are mean +/- SEM Values are normalized against endogenous control gene

M-mouse H-human

Workinprogress:
MagnetizationTransferisanMRItechniquetosensemacromolecules. Canweuseittoimagethefibrousstroma,andifso,tomonitor hedgehoginhibitorsinpancreaspatients?

Inefficientangiogenesisresultsin tumourhypoxia
longknowntoassociatewithmetastaticpotentialand adversepatientoutcome

IntJRadiat Biol Oncol Phys2000;48:91922

Eppendorf probe

HypoxiainPancreaticCancer
Occurswhenoxygenconsumption exceedssupplybyvascularsystem Inferredtooccurinpancreaticcancer patients,basedonlowperfusionusing intravenouscontrastagents Directtumour pO2 measurements

Hypoxiaactivatesnitroimidazole probes
TISSUE FIXATION OF EF5 UNDER HYPOXIA N N R NO2

e-

REACTIVE PRODUCTS

Bindtointracellularmacromolecules BIND TO R=CH2CONHCF2CF3 antibodydetection

O2

.-

O2

MACROMOLECULES ANTIBODY DETECTION

Surgical sample

Orthotopic xenograft

100m EF5 BrdU

EdU

EdU EF5

EF5

Consistentresultsfromreplicatetumours indicategeneticorepigeneticmechanism(s) involved

Hypoxiatracksincreased BrdU labeling andmetastaticpotential

Hypoxiaandcellcyclearrestoccurearlyin thegrowthoflivermetastases
implicationsforadjuvantgemcitabine
EF5 BrdU DAPI

200m

Isthisclinicallyrelevant?
Clinicaltrialofhypoxiaprobepimonidazole

Pimonidazoletrial
Withoralformulation,expecttoaccrue~40 patients/year Preliminarydatasuggeststainingpatternsandrangeof hypoxiasimilartoxenografts Hypoxiainearlystagepancreascancer: colocalizetomarkersofdifferentiation, invasiveness,stemness etc. doprimaryxenograftsrecapitulatepatient? underlyinggeneticmechanisms Doeshypoxiapredictearlyrecurrence? ifso,willneedtochangeclinicalpractice

Whatifhypoxiccancersdobadly?
(Imean,worsethanusualforpancreasca!)

Hypoxiccellspredictedtoberesistanttocell cycleactivedrugslikegemcitabine,but sensitivetoagentsthatundergobioreductive activationunderhypoxia

TH302activation

Preliminarydata TH302istoxictopancreas cancerstemcells,usinglimitingdilutionassay

OCIP 51 control 10Gy TH302 TH-302 + 10Gy TIC frequency 1:4 1 : 1386 1 : 107 1 : 4813 SE 0.3652532 0.3726 0.3272 0.3446

Irradiatingtumours growninmouseleg

Measuringhypoxiainunresectable or metastaticpancreascancer

F18FAZA

Preliminarydatausingprimarypancreascancerxenografts usetheseinordertostandardizeinterpretationandreporting ofFAZAinpatients

FirstclinicalimagesofFAZAPETin cervixcancerpatients

Patient1

Patient2

Blurringtheboundariesbetweensurgical, medical,andradiationoncology
Resectable pancreascancerpatients