This document summarizes non-invasive small animal imaging modalities used in cancer drug research and development. It discusses bioluminescence, fluorescence, PET, ultrasound, and CT imaging and their applications and characteristics. It then profiles the P-cadherin target and shows how a P-cadherin IgG displays antitumor and antimetastatic effects in multiple cancer models through bioluminescence, fluorescence, PET imaging and immunohistochemistry. Fluorescence imaging also showed the IgG distributed to subcutaneous and subrenal capsule tumor sites.
Non-invasive small animal imaging reveals antitumor and antimetastatic effects of PF-03732010 in preclinical cancer models
1. Non-invasive small animal imaging in cancer drug
research and development
Cathy Zhang, Translational Research Group, Oncology Research Unit, Pfizer
Global Research and Development, La Jolla Laboratories
2. Preclinical imaging modalities overview
Clinical
Modalities Light Output Application Characteristics
Imaging
anatomical, high throughput, low cost
BLI luciferase labeled cells
functional, high sensitivity/ No
Bioluminescence /luciferin injection
molecular low resolution
fluorescence labeled anatomical, high sensitivity
Fluorescence molecules or cells functional, Limited
(exogenous/intrinsic) molecular (low cost)
18FLT-PET injection of 3’-Fluoro-3’ anatomical, high resolution Yes
deoxythymidine functional (high cost)
interaction of sound anatomical, high resolution Yes
Ultrasound
waves with living tissues functional (vasculature)
anatomical, high resolution Yes
CT n/a
functional (bone, lung)
Target – P-cadherin, CDK4 and Chk1
3. P-Cadherin target key profile
Cell 1 Cell 2
Intracellular
Plasma Space Cell surface glycoprotein /involved
Membrane in Ca++-dependent cell-cell
adhesion.
actin
Cadherin cytoskeleton Signalling is mediated by β-
Dimers catenin
nucleus In the cytoplasm, β-catenin is
Adherens stabilized by Wnt signaling and
Β-catenin/Tcf
Junction translocates to the nucleus
Wnt Target gene
transcription In the nucleus, β-catenin/partners
w/ Tcf factor to regulate
Cytoplasm expression of oncogenes such as,
c-myc, survivn, and cyclin D1 etc.
tissue cell
integrity Proliferation
cell
invasion
4. Disease progression in MDA-MB-435-HAL SRC model
Fluorescence Bioluminescence
day 33 IVIS200 (Caliper)
ProSense
Injection
FMT (VisEn)
Normal Lung
14 28 36 55
Days Post Tumor Implant
16
BLI (photons/sec x 108)
100 100
% of Survival
12
Percent survival
SRC Model 75 Primary Tumor Mouse 75
Growth Survival
Tumor Burden
8
50 in Lungs 50
4 25 25
0 0 0
0 25 50 75 100
0 25 50 75
Days Post Tumor Implant 100
Primary tumor and lung metastasis can be longitudinally tracked via BLI and FLI
5. PF-03732010 displays antitumor and antimetastatic property
MDA-MB-435HAL-CDH3-SRC PC3M-CDH3
Lymph Node
Metastasis
Dorsal Vehicle
Primary
Vehicle PF-2010 Tumor
(20 mg/kg)
PF-03732010
20 mg/kg
Ventral
CTC (hAlu)
PF-03732010 (mg/kg)
20
H&E 5
Staining
1
0 50 100 150 200
% of Control
PF-03732010 induces antitumor and antimetastasis property in multiple models
6. Tumor growth inhibition of PF-03732010 in SC and SRC Model
MDA-MB-435HAL-CDH3
16
1000 Control
BLI x 108 (photons/sec)
PF2010, 20mg/kg
120
Tunor volume (mm3)
Vehicle
12 PF-2010, 40 mg/kg 800
Relative Tumor Size
PF-2010, 20 mg/kg 90
(% of Control)
PF-2010, 10 mg/kg
600 SC
8 SRC 60 implant
implant 400
30
4
0 200
SRC SC
Vehicle PF3732010
0
10 20 30 40 50 60 10 20 30 40 50 60
Days Post Tumor Implant Days post tumor implant
PF-03732010 displays better efficacy in the MDA-MB-435HAL-CDH3 SRC model than
SC model
7. Fluorescence imaging detect the IgG (PF-03732010) distribution
10 IgG Distribution
M1 M2 M3
(x 10 photons/sec)
8
24 hr
1 hr
Intensity
6 72 hr
4
9
24 hr 2
SRC tumor 0
sc tumor
y
ng
n
or
nt
r
or
ve
ne
SR Ski
.I
m
m
Lu
Li
id
Sm
Tu
Tu
72 hr
K
SC
C
Serum concentration, nM
1000
IVIS200 (Caliper) T1/2 = ~ 7 days
CL = 0.01ml/min/kg
Vss = 0.18 L/kg
Tumor implanted in sc (right flank) and 100
subrenal capsule (left side)
SC injection with P-Cad IgG
10 mg/kg IV
Drug level reached Cmax in 24 hr 10 10 mg/kg s.c.
IgG distributes to sc and SRC location
0 168 336 504 672
Time, hr
8. PD modulation with the treatment of P-Cadherin IgG
β-Catenin Merged Caspase 3 Ki67
IHC –
Vehicle
β-Catenin change - target
modulation
PF-2010 Increased apoptosis &
20 mg/kg
decreased proliferation
18FLT-PET
[18F]FLT-PET/CT CT Imaging
[18F]FLT-PET Imaging
Vehicle Vehicle Vehicle
Suppression of [18F]FLT uptake
kidney
tumor
PF-2010 PF-2010 PF-2010
Tumor in SRC Tumor in Lung Day 42
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