wp6. cancer imaging with focus on breast cancer...wp6. cancer imaging with focus on breast cancer...

Wp6. Cancer Imaging with focus on

bbreast cancer

Anikitos Garofalakis

‘Frederic Joliot Hospital Service(SHFJ), Institute of Biomedical

imaging(I2BM), Atomic Energy Commission(CEA)’g g( ) gy ( )

1Anikitos GAROFALAKIS CEA, I2BM, SHFJ, LIME, INSERM U1023

background

The use of animal models for the evaluation of FMT under realistic conditions

Objectives of Wp6:

To provide cancer animal models

To provide key fluorescence probes

Quantitatively examine FMT performance to visualize disease processes in-vivo

To predict clinical utility (48 month deliverable)


Animal modelsmodels and probes

MCF 7MDA 231 U87PC12 MEN2A

a. xenografted tumors

PyMT

b. spontaneous tumors

Animal models

MCF-7MDA-231 U87 human glioma

PC12-MEN2A rat pheochromocytoma

PyMT

Fluorescent probes

Prosense RGD-based Angiosense

a. commercial probes

Aptamers Nano-micelles

b. custom-made probes

Prosense RGD basedoptical probes(Integrisense680,Angiostamp680)

Angiosense


Cathepsin activity Integrin avβ3 Blood volume

fluorescent probes for tumor-related processes

FDG

Integrin avβ3 Neo-angiogenesis

CathepsinECM d d tiECM degradation

Blood volume


Quantitatively examine FMT performance to visualize disease processes in-vivo

targetedbiological molecule 1

FDG

μPET FMT

Type of information Molecular Molecular

Resolution ~ 1 mm3 ~ 1 mm3

Kinetics Fast (~seconds) 5-15 min/scan

Whole Body Imaging YES NO

Time range of follow up ~ hours ~days

Cost $$$ $Cost $$$ $

Activatable probes NO YES

Labelling Need radiochemistry Simple

Quantification Linear from pM ?


Q p ?

calibration of FMT

Use of tubes filled with fluorophores

Use of the quantification capacity of PET imaging in combination with a dual PET/Optical probe

targetedbi l i l biological molecule

PET

vs [C]

FMT


FMT-PET-CT imaging

in vivo


fDOT-PET-CT imaging/protocol

PET PETFMT CT

0 60 min30 90

CT contrast agenttargetedbiological molecule


calibration/FMT-PET-CT imaging

PET FMT PET CT

FMT/CTul

ated

es

) 800

1000

quan

tity

calc

y PE

T (

pmol

e

600

PET/CT

Prob

e by

200

400 PET/CT

Time (min)

0 20 40 60 80 100 120 140


b/./calibration/fDOT-PET-CT imaging/volume validation

PET/CT fDOT/CT

Coronal view

Axial viewAxial view

PET segmented volume

Optical signal segmented volume

CT contrast


CT contrast

b/./calibration/fDOT-PET-CT imaging/quantification

1000

(M

) 10

oles

by

PET

10

100

tion

by P

ET

0,1

1

pm

o

0 1

1

Con

cent

rat

0 001

0,01

fDOT (a.u.)101 102 103 104 105

0,1

fDOT concentration (a.u.)0,1 1 10 100

0,001

Sensitivity limit ~ 1 pmoles, 5nM(C)


Garofalakis et al, Optics Letters (2010)

PET-FMT dual imaging

targetedbiological molecule

μPET FMT

Type of information Molecular Molecular

Resolution ~ 1 mm3 ~ 1 mm3

Kinetics Fast (~seconds) 5-15 min/scan

Whole Body Imaging YES NO

Time range of follow up ~ hours ~days

Cost $$$ $Cost $$$ $

Activatable probes NO YES

Labelling Need radiochemistry Simple

Quantification Linear from pM Linear from nM


Linear from nM

cancer imaging in combination with PET

Angiostamp680ControlAngiostamp680Angiosense750

FMT PET CT FMT

day -1 day 0 day 1 day 2 ….

Prosense680


b

MDA-231/FDG-Cathepsin

a b

3D Optical( )

0.92

0.75 mm(Prosense)

0.59μM

PET [F18]FDG

c

Volume Mean

Prosense_DAY_1 317.781 3.5




MDA-231/FDG – Integrin avβ3

RGD-based ligand Control ligand

0.24

0 12

0.04

0 02

0

0.12

uM0

0.02

uM


models and probes

MCF-7MDA-231 U87 human glioma

PC12-MEN2A rat pheochromocytoma

a. xenografted tumors

PyMT

b. spontaneous tumors

human gliomarat pheochromocytoma

Probe to tumor

MDA231/ Cathepsin

activity

MDA231/ Integrin

localization

MDA231/ Nano-micelle labelling

PC12-MEN2A/Cathepsin activity

PC12-MEN2A/Cathepsin activity/tumor

% signaloverlapping 6,6 ± 2,5 6,7 ± 5,0 34 ± 12 31,8±12,0

p yIntegrin localization

% volumeHMGU@MunichD l 7 4% volume

overlapping 37,6 ± 8,1 39,0 ± 18,6 40,03 ± 19,02 41,2±12,5 Del 7.4


MCFS7 xenografted tumor/aptamers ACE8

0.80.9

0 30.40.50.60.7

D in

tum

our

“Annexin II” as potential target ?

ACE8 Control0.00.10.20.3

%ID

*


summary

To provide cancer animal models

Del 6.3 To develop animal models of other cancer for studying FMT-XCT performance (U87 glioma cells)

To provide key fluorescence probes

2nd year deliverable (target of aptamer ACE8 has to be identified)2 year deliverable (target of aptamer ACE8 has to be identified)

Quantitatively examine FMT performance to visualize disease processes in-vivop

Del 6.4 To perform in-vivo imaging of key animal models of cancer and correlate the findings with standard laboratory tests and gro th meas resand growth measures

To predict clinical utility (48 month deliverable)


aknowledgements

Albertine Dubois Dr

Laboratory of experimental molecular medicine (LIME)

Albertine Dubois, DrXiao Tong, Phd studentVincent Brulon, TechnicianRaphael Boisgard, Dr

Chemistry Laboratory for life sciences(LCV)Bertrand Czarny, Techniciang

Bertrand Tavitian, ProfFrédéric Ducongé, Dr

y,

Group of radiochemistry et radio-pharmacy (GRR) Bertrand Kühnast, DrFrederic Dollé Drex-members:

Agnes Cibiel, DrCarine Pestourie, DrDaniel Dupont, Dr

Frederic Dollé, Dr

Nicolas Mackiewicz, DrIsabelle Jassens, Technician


a/fDOT-alone output

xyz

x

PC12-MEN2A with SentiDye® 700


coronal sagittal

a/fDOT with CT

Tumor model MEN2A/Integrisense(angiogenesis)

fDOT is often combined with a structural modality


Unmixed Unmixed

b/./cancer imaging in combination with PET/FDG-Two color fDOT

a b

UnmixedAngiostamp680

UnmixedAngiosense750

5.0

11.0

0.9

7.7

c dCT/Optical CT/Optical

μM μM

11.0 4.4

[F18]FDG segmented volume

Angiostamp680 signal segmented volume

FDG/Optical FDG/Optical

5.0μM

0.9μM

Angiosense750 signal segmented volume

e f g

h i j


b/./calibration/fDOT-PET-CT imaging/protocol

PET PETfDOT CT

0 60 min30 90

Kidney contrastagent

Oligo

+Oligo

+

Known biodistributionKnown biodistribution

Deep seated organ


No degradation

b/./calibration/fDOT-PET-CT imaging

PET/CT fDOT/CT


b/./calibration/fDOT-PET-CT imaging/volume validation

a) b)

PET segmented volumePET segmented volume

Optical signal segmented volume

CT contrast

PET/CT fDOT/CTPET/CT fDOT/CT


b/./calibration/fDOT-PET-CT imaging/quantification

1000 10

calc

ulat

ed

mol

es) 100

robe

cal

cula

ted

M)

1

robe

qua

ntity

cby

PE

T (p

m

1

10

entr

atio

n of

pr

by P

ET

(

0,01

0,1

Reconstructed fDOT fluorescence (a.u.)

101 102 103 104 105

P

0,1

Reconstructed fDOT fluorescence concentration (a.u.)

0,1 1 10 100

Con

c

0,001

Reconstructed fDOT fluorescence (a.u.) ( )

Limit ~ 1 pmoles, 5nM(C)


a/fDOT imaging

hCd2-GFP mouse

Garofalakis A et alThree-dimensional in vivo imaging of green fluorescent protein-expressing T cells in mice with noncontact fluorescence molecular tomography. Mol Imaging. 2007 Apr-Jun;6(2):96-107


Diffuse light in a homogeneous medium

continuous wave point source

DSUU )()()( 022 rrr

D

p

D D

l i f h d i J

+

solution for the energy density Jn

+ SourceeUr

)(rr

)(


Diffusive semi – infinite medium

Integrisense 0h Integrisense 3h

b/./MDA-231/Integrisense and AngioStamp comparison

gIntegrisense and AngioStamp comparison

g

Tumor volume ~ 730 mm3

60,2 % signal of optical in the surrounding

30,01 % volume of tumor in the common area



RAFT-RGD 0h RAFT - RGD 2h

Tumor volume ~ 670 mm3






Discretization and weight matrix

• Sample the fluorescent concentration in N points

r1 r2 r3N points(voxels)• Convert integral equations into a system of linear equationssystem of linear equations

rny

x

For M projections : P[M×1] = W[M×N] · N[N×1]

ART method


ART method

b/./cancer imaging

Commercial probes

P 680Cathepsin Prosense 680Cathepsinactivity

Integrinsense680 (VisenMedical, USA)AngioStamp (Fluoptics, France)

Integrin localization

Custom-made probesCustom made probes

MPEG nano-micelles

Passivebinding of tumors


b/./calibration

Use of fluorophores of controledconcentration

Use of the quantification capacity of PET imaging in compination with a dual PET/Optical probe

vs [C]

L t th d b t t li ti Hi h t th d b t li ti


Low cost method but not realistic High cost method but realistic

wp6. cancer imaging with focus on breast cancer...wp6. cancer imaging with focus on breast cancer...

Documents