The presentation is an introduction to using bioluminescent "reagents" to evaluate drug efficacy in tumor models. This presentation briefly highlights one of many research platforms available at Caliper Life Sciences\' Discovery Alliances\' In Vivo Division that performs contract research for the life science community.

1. Bioluminescent Imaging for Oncology Studies
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2. Bioluminescent/Fluorescent Imaging
Introduction
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3. Advances in Biology:
Probe and Model Development and Validation
Luciferase
Fluorescent
Proteins
Fluorescent Probes
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4. Integration with Biophotonic Imaging Assays
Image
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5. Current vs. IVIS Methodology
Conventional Methodology = 24 animals are sacrificed over four time points
Week 1 Week 2 Week 3 Week4
IVIS Methodology = 6 animals over four or more timepoints
Week 1 Week 2 Week 3 Week4
The same group of animals is reused at each time point; far fewer animals
are used in this experiment than using conventional technologies
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6. Oncology Techniques
Caliper measurements Non-invasive bioluminescent
imaging of implanted light
Only for S.C. models
producing tumor cell growth in
Limited to solid tumors
the following models:
>100 mm3 tumor size
– Subcutaneous
Tumor cell line does not require
genetic manipulations – Orthotopic
– Metastatic
Histopathology More sensitive: as few as
100/500 cells can be detected
Time consuming
(depending on reporter system)
Terminal procedure
Light correlates with the number
Requires a lot of animals of live cells in the tumor
Tumor cell line does not require
genetic manipulations
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7. Applications of IVIS Technology
Oncology
Infectious Diseases
Immunology, Inflammation and Autoimmune Diseases
Cardiovascular
Diabetes and Obesity
Neurology
Drug Metabolism
Gene Therapy Vectors
Formulations & Biodistribution
Stem Cell Research
Gene Therapy
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8. Therapeutic Area Specific - Models and Applications
Oncology - Spontaneous tumor model Inflammation – Arthritis model
CNS - Stroke Models
EL1-Tag- Luc Fluorescent conjugate detects
GFAP-Luc
Thrombin activity
Stem Cells – grafting of
Primate Models – Viral gene transfer
Metabolic Disease – Diabetes model
Neuronal stem cells
Macaque Model
RIP-Luc
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9. Xenograft Models
Human Tumor Cell Lines in Immuno-
deficient Mice
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10. Light Producing Human Tumor Cell Lines
Constitutively expressing luciferase or a fluorescent protein used in
primary and metastatic tumor growth models.
Lung Prostate
- A549-luc-C8 - PC-3M-luc-C6
- LoVo-6-luc-1 - LNCaP-luc-M6
Breast Colorectal
- MDA-MB-231-luc-D3-H1 - HT-29-luc-D6
- MCF-7-luc-F5 - LoVo-6-luc-1
- MDA-MB-435-luc-G2
Ovarian Cervical
-SKOV3-luc-D3 - HeLa-luc
Customized light producing tumor cell line production also available.
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11. Example of an Oncology Study
Light producing (luciferase or fluorescent protein expressing) tumor
cells are implanted into immuno-compromised or immuno-competent
mice/rats.
The tumor growth/metastasis formation is monitored once/twice a week
using none-invasive in vivo imaging (IVIS) technology.
Compound administration is initiated when tumor(s) are established
based on the results of preliminary IVIS imaging.
The efficacy of the compound is monitored by imaging using IVIS
instrumentation and/or caliper measurements.
Additionally, we have capabilities to perform toxicity and
pharmacokinetic studies prior to conducting the efficacy study.
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12. Subcutaneous Tumor Models
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13. Subcutaneous Tumor Models
PC3M-luc-6
Human Prostate Therapeutic Treatment Paradigm
Adenocarcinoma
Day 14 Day 21 Day 28 Day 35
No Treatment
Mouse #6
2.2x109
n=7
Mouse #40
n=8
5-FU
(treatment started
2.8x108
on Day 11)
Mouse #31
Mitomycin C
n=8
(treatment started 3.6x106
on Day 11)
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14. Subcutaneous Tumor Models
Drug Analysis In Vivo – PC-3M-luc 1x106 Cells s.c.
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15. Orthotopic Tumor Models
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16. Orthotopic Models for Prostate and Breast
Cancers
Technique:
Orthotopic injection of
Tumor cell suspension
MDA-MB-231-luc-D3H1
Human Mammary Gland Adenocarcinoma (pleural effusion)
PC3M-luc-6
Human Prostate Adenocarcinoma
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17. Orthotopic Colon Carcinoma Model
LoVo6-luc-1
Human Colorectal
Adenocarcinoma
S.C. tumor excision
Tumor fragment
Orthotopic model
transplant surgery
Week 5
Day 0 Week 6 Week 7
Week 4
Compound Treatment
Subcutaneous model
Measurements:
Bio-luminescence
Endpoints:
Day 0 Week 1 Week 2 Week 4
Tissue collection for histopathology
and biochemical analysis
Measurements:
Bio-luminescence
Tumor volume (caliper)
S.C. Injection
In left flank
week 4 week 7
Luminescent
LoVo-6-luc cells in vitro
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18. Orthotopic Prostate Tumor Model
Therapeutic Treatment
Disease Relapse
Week 1 Week 3 Week 5 Week 7 Week 8
n = 13
Saline
Control Group
Mouse #40
End Point
iv, 3x/wk — 100
— 100
Week 5
Wk 2, 3
— 80
— 80
n = 6/7
5-FU
x106
x106
— 60
— 60
Mouse #38
100mg/kg, iv, 1x/wk — 40
— 40
Wk 2, 3, 4
— 20
— 20
Mitomycin C n = 3/5
Mouse #4
2/mg/kg, iv, 3x/wk
Wk 2, 3
PC-3M-luc-C6, 5x105 Cells; Male nu/nu CR
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19. Metastatic Tumor Models
Experimental and Spontaneous
Metastases
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20. PC3M-luc-6
Experimental Metastases
Human Prostate
Intravenous Models
Adenocarcinoma
nu/nu
SCID-bg
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21. PC3M-luc-6
Experimental Metastases
Human Prostate
Intracardiac Models
Adenocarcinoma
nu/nu
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22. PC3M-luc-6
Spontaneous Metastases
Human Prostate
Subcutaneous Model
Adenocarcinoma
Primary Tumor Primary Tumor is Shielded
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23. Spontaneous Metastases
MDA-MB-231-luc-D3H1
Human Mammary Gland
Orthotopic Model
Adenocarcinoma
(pleural effusion)
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24. Metastatic Tumor Models
Inhibition of Tumor Growth by SU11248
Experimental Breast Cancer Bone Metastasis Model
Murray et al 2003
435/HAL-Luc Cells
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25. Bioluminescent/Fluorescent Imaging
Success Stories
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26. Caliper Technology Played a Role in the Preclinical
Development of the Following Therapies
FDA Approved In Clinical Trials
Pfizer, Sutent (sunitinib) Phase I:
(kidney and stomach cancer)
Cell Genesys, CG0070
Novartis, Tasigna (nilotinib)
(Bladder and multiple indications)
(CML, chronic myeloid leukemia, Gleveec resistance)
Nereus Pharma, NPI-0052
Novartis, Zometa (zoledronic acid) (multiple myeloma)
(metastasis of breast, lung, prostate and multiple myeloma)
Novartis, AEE788
BMS, Sprycel (dasatinib) (Advanced Cancers)
Millennium, PS-341 (combo)
(CML, chronic myeloid leukemia, Gleveec resistance)
(Non-Hodgkin’s Lymphoma, others)
Insert Therapeutics, IT101
Cubist Pharma, Cubicin (daptomycin)
(solid tumors)
(S. aureus infections – MRSA treatment)
Novartis, CHIR-258
(Metastatic melanoma)
Phase II,III:
Sanofi-Aventis, Aflibercept
(Multiple indications)
EntreMed, Panzem
(Recurrent Glioblastoma)
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27. Examples of Recent Oncology Drugs
IVIS used in Preclinical Studies
IT-101: Phase I clinical trials to
Tasigna is an FDA approved
AEE788 (Novartis) is in Phase I
determine its safety and toxicity for
drug for the treatment of
clinical trial for patients with
treatment of patients suffering from
Philadelphia chromosome
Advanced Cancer
Advanced Solid Tumors
positive chronic myeloid
leukemia (CML)
Schluep et al., Clin Cancer Res
Weisberg et al., Cancer Cell :
2006;12(5)March1, 2006, p. 4908–4915
Lu et al., Clin Cancer Res 2007;13(14) Feb. 2005 — Vol. 7, p. 129-141
July 15, 2007, p. 4209–4217
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28. Business Development Contacts
Alex Chang, Ph.D.
Associate Director, Business Development
E-mail: alex.chang@caliperls.com
Phone: 609-235-1413
Mobile: 609-969-8019
Stephen J. McAndrew, Ph.D.
Vice President, Business Development
E-mail: stephen.mcandrew@caliperls.com
Phone: 609-235-1420
For more information about the Discovery Caliper Life Sciences
Alliances & Services Division, please visit Discovery Alliances & Services
www.caliperLS.com . Division (Xenogen Biosciences)
5 Cedar Brook Drive
Cranbury, New Jersey 08512 USA
Phone: 609-860-0806
Fax: 609-860-8515
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