S. Di Giovine
Business Development Director
29 Novembre 2011
Università degli Studi di Milano
Via Golgi 19 aula G10
Corporate profile
TOP srl, originated in 2006 as a spin-off of
the University of Milan and has been backed
in 2007 by the NEXT fund managed by
Finlombarda Gestioni SGR.
TOP is located at the Parco Tecnologico
Padano in Lodi, near Milan.
TOP research team is specialized in the generation
of vectors, cells and animals for Molecular Imaging
using:
 Bioluminescence
 PET (Positron Emission Tomography)
 NMR (Nuclear Magnetic Resonance)
TOP mission
To provide the scientific community with non-invasive tools for the study of
the activity of drugs and environmental or food contaminants
TOP aims at combining the power of physics, informatics and molecular
biology to generate novel models for the bio-medical research and for
the control of the environment and alimentary chain.
TOP strives to renew present investigational tools in the respect of animal
rights to satisfy two conflicting needs: use of predictive biological models
in the absence of animal distress.
The Company expertise: Molecular Imaging
Molecular imaging is based on the appropriate use of REPORTERS: molecules
easily detectable and allowing to study spatio-temporally molecular events in living
organisms.
Ligands
Ligand
NR
NR
NR
NR
Reporter
green fluorescent dye
The Company unique and proprietary technology*:
the use of insulator sequences to obtain generalized
expression of the reporter
CCD
Camera
* EU patent No. EP 1298988B1 US patent No. 7,943,815
All TOP products are subjected to intense
validation to ensure:
PREVENTION OF EPIGENETIC MODIFICATIONS
REPRODUCIBLE and REGULATED EXPRESSION OF THE REPORTER
REPORTER EXPRESSION MAINTAINED IN TIME AND GENERATIONS
constitutive/ inducible
SATURN
VALIDATION
ACCURACY
SPECIFICITY
TURN-OVER
UBIQUITY
REPRODUCIBILITY NON-INVASIVENESS
TOP products enable to study your favorite molecular
process along the whole life cycle
E 19.5
P.N. 1
E 18.5
18.5
P.N. 10
E 17.5
16.5
E 16.5
15.5
DAY 14.5
E 14.5
14.5
E 12.5
Embryonic Dev. & Pregnancy
3 months
Adulthood
TOP finds a response to a variety of research needs
 Pharma:
−
−
−
−
−
−
−
identification of novel drug targets
speeding of pre-clinical testing
identification of drugs endowed of tissue specificity of action
more predictable toxicological tests
gender pharmacology
pharmacology of infancy and aging
new indications for drugs in the clinic or in the market
 Environmental & Food toxicology:
−
−
−
−
−
identification of specific activities in mixtures of compounds
threshold effects in short and long term exposures
identification of food contaminants
threshold effects in short and long term exposures
identification of the organs targeted by environmental pollutants
 Biomedical research:
− understanding the physiological effects of specific molecular events
One single tool for the multidimensional analysis of
drugs in vivo.
 Immediate readout;
 Increased prediction power:
- holistic vs. reductionist approach;
- drug action, not only distribution.
 Increased ability to perform multidimensional
analysis of drug activity:
- space (organs, PD);
- time (PK, longitudinal studies);
- immediate evidence of undesired activity
(toxicology).
TOP Reporter
system
 Lower costs;
 Decreased research time;
 Significantly reduced number of animals.
Partnership
TOP srl is partner of Caliper Life Sciences (former Xenogen) , the
world leader for in vivo optical imaging.
TOP reporter models - I
 repTOP™ mice available at present:
Models
Therapeutic areas:
repTOP™ ERE-Luc mouse
(Estrogen Receptors)
•Aging-related pathologies
•Depression
•Inflammation
repTOP™ PPRE-Luc mouse
(PPAR’s)
•Lipid metabolism alteration
•Diabetes
•Cancers and others
repTOP™ MITO mouse (“MITO as in MITOSIS”)
•Oncology
•Regenerative medicine
•Toxicology
repTOPTM Ubi-Luc2 (series) mouse + VivoGloTM
• Metabolism (CYP3A4)
• Apoptosis (Caspase 3/7)
TOP reporter models - II
 repTOP™ mice soon available
Models
TM
repTOP
TM
repTOP
TM
repTOP
TM
repTOP
Therapeutic areas:
NFkB-Luc2 mouse
TRAIL-EGFPLuc mouse
P21-Luc /EGFPTK mouse
ARE Luc2
Apoptosis
Inflammation
Proliferation
Oxidative stress
In addition, TOP has generated a vast array of constructs for
cell and animal engineering ready to be used and addressing
most of current major needs in biological research. All of
these constructs contain two or more reporters enabling
bimodal and trimodal applications of molecular imaging
TOP reporter models - III
Custom- made repTOP™ mice: TOP’s technology platform and the
standardized validation process are the guarantee of a rapid and reliable
generation of an industry standard-compliant reporter system.
Feasibility
4-5 weeks
Generation
Characterization
15 -18 weeks
Validation
Founder mouse
OUTSOURCING YOUR RESEARCH AT TOP
The expertise of TOP research team in molecular imaging is at our
client’s service for advise and testing of compounds in a wide variety of
therapeutic areas
TOP mice in the world
 Italy:
 USA:
− Spedali Civili, Brescia
− University of Kansas Medical Center (KUMC)
− Roma, University Tor Vergata
− University of Miami, University of Chicago
 Europe:
− Institut de Genetique et de Biologie Moleculaire
et Cellulare (IGBMC)
− University of Illinois
− UT Southwestern Medical Center at Dallas
− University of Pennsylvania School of Medicine
− Karolinska Institute
− University of Tennessee
− Max Planck Institut fuer Psychiatrie Muenchen
− Dana-Faber Cancer Institute
− Dartmouth Medical School
− Baylor College of Medicine.
− Leiden University Medical Center.
 Australia:
− Prince Henry’s Institute of Medical Research
 Leading Pharmaceutical and
Biotechnology companies worldwide.
TOP newest product:
repTOPTM mitoIRE to study proliferation in vivo
The repTOPTM mitoIRE mouse to study
proliferation in vivo
Anti – proliferative stimuli
Pro–proliferative stimuli
CCD
NF-Y promoter
1u
Cyclin B2
minimal promoter
Cam
era
repTOPTM mitoIRE validation
Photon emission
(p/s/cm2/sr)
cell proliferation induced by application of 7,12-dimethylbenz(a)anthracene
to the skin of the left ear can be easily measured in vivo
150000
100000
50000
0
0d
1d
2d
Time (days)
3d
6d
9d
COMPARING the sensitivity of Ki-67 immunohistochemistry and
bioluminescence in the MITO mouse
Cell Proliferation
(p/s/cm2/sr)
0,3
0,2
0,1
0
d3
d9
Imaging
d9
vehicle
Bioluminescence:
low background and
rapid detection
DMBA
Cell Proliferation
(pos nuclei/tot nuclei)
d3
DMBA
vehicle
Ki-67 immunoistochemistry
d3
d9
80000
60000
40000
20000
0
d3
d9
repTOPTM mitoIRE
following in time the development of chemically
induce prostate carcinoma
Photon emission
(cts/sx103)
200
100
12
4
1
0
TM
TM
Combining repTOP and VivoGlo technologies
repTOP™ UbiLuc2-M8 mouse
Z-DEVD-Aminoluciferin, Sodium Salt
Induction of liver apoptosis
6h i.p. treatment with:
D-GalN (800mg/kg) + LPS (100 µg/kg )
Nakama T et al. Hepatology. 2001