La ricerca di base incontra l`industria per la cura delle malattie

Il cervello:
La ricerca di base incontra l’industria
per la cura delle malattie
Laura Cancedda, Ph.D.
Neurodevelopmental Physiology Lab.
What Can I Do for You?
Basics about brain development and function
Overview about experimental techniques
Challenges for development of SNC medicines
The Human Brain Is Responsible For Generating
Behaviors That Promote the Welfare of an Animal
Adult brain 1,300 - 1,400 g
Roughly 100 billion neurons
linked with up to 10,000 synaptic connections each
The Brain is Organized in Structurally and
Functionally Distinct Areas
Distinct Brain Areas Are
Interconnected Among Them
The Neocortex is a Very Organized Structure
How Is This Complexity
Generated?
DEVELOPMENT
Intrinsic Programs
Eye Opening
Young (Human)
Berardi, Pizzorusso 2007
PLASTICITY
Sensory Experience
Odor
stimuli
Visual
Stimuli
Acustic
stimuli
Tactile
stimuli
Adult (Human)
SNC Development
Neuronal plate
Neuronal tube
(embryonic ectoderm)
Neuronal tube
All neurons originate from the same place
Many Fundamemental Processes of Development
Are Affected in Neurodevelopmental Disorders
Down Syndrome
Schizophrenia
DiGeorge Syndrome
Autism
Rett Syndrome
Fragile X Syndrome
Tourette Syndrome
Epilepsy
Cerebral Palsy
Neurofibromastosis
Distinct Brain Areas Are
Interconnected Among Them
Neuronal Transmission
Axon
Neuron A
Axon Neuron A
Dendrite
Synapse
Dendrite Neuron B
Neuron B
GABA and Glutamate Regulate Brain Activity
GABA
E
Glutamate
I
GABA and Glutamate Together
Regulate Brain Activity
How Can We Study Neurons?
(1) Visualization
Green Fluorescence Protein (GFP)
is Used to Visualize Neurons
The 2008 Nobel Prize in Chemistry Was Awarded
to the Inventors of GFP
Roger Y. Tsien
Martin Chalfie
Shimomura Osamu
Brainbow
20µm
Healthy Brain
Diseased Brain
Neuronal Transmission
How Can We Study Neurons?
(2) Electrophysiological Recordings
The Patch-Clamp Setup
Healthy Brain
4-AP 200µM
2
3
0.3
2
3
0.3
0.2
LII/III
0.2
0.1
0
IN 1
(mV)
IN 1
(mV)
0.1
-0.1
0,2 mV
-0.2
0.3
-0.2
0.3
0.2
0.2
5 min
0.1
IN 0
(mV)
0.1
0
LV
IN 0
(mV)
0
-0.1
0
-0.1
-0.1
-0.2
-0.2
3700
35
40
45
50
55
60
65
3720
3740
3760
70
3780
3800
3820
3840
Time (s)
Time (min)
Diseased Brain
4
2
3
3
0.3
0.2
0.2
LII/III
0.3
IN 0
(mV)
0.1
0
0
-0.1
-0.1
-0.2
-0.2
0.3
0.3
0.2
0.2
LV
IN 0
(mV)
0.1
0.1
IN 1
(mV)
IN 1
(mV)
0.1
0
0
-0.1
-0.1
-0.2
-0.2
15
20
25
30
35
Time (min)
40
45
50
10s
2520
2540
2560
2580
2600
Time (s)
2620
2640
2660
Down Syndrome
Mouse models
Human
Antonarakis et al., 2004
How Can We Study Neurons?
(2) Behavioral studies on animal models
Healthy Mouse
Multisystem:
- cardiac defects
- alterations of the craniofacial skeleton
- immunological defects
- smaller size
Brain:
- increased susceptibility to seizures
- mild Alzheimer-like degeneration
Down Syndrome
mouse
Down Syndrome Animals are Hyperactive
Healthy Mouse
Down Syndrome
mouse
Cell-attached configuration
Bumetanide Can Restore
Inhibitory GABA Transmission
Cell
Patch
pipette
Cell-attached configuration
GABA is Excitatory
in Adult Ts65Dn Mice
Cell
Patch
pipette
Deidda et al., Nature Medicine, 2015
Bumetanide
0.2 mg/Kg/Day (i.p.)
0
1
2
3
4
Weeks
TEST
TEST
TEST
Object Location
Novel Object Recognition
Contextual Fear Conditioning
24 h
24 h
24 h
Shock
***
20
-40
-100
Provisional Application: US 61/919,195, 2013
90
30
-30
-90
**
**
100
Time freezing (%)
***
Discrimination index
Discrimination index
80
***
Context test
***
50
0
Deidda et al., Nature Medicine, 2015
Expression of NKCC1 is Increased
In Adult Ts65Dn Mice
and Down Syndrome Patients
WT
Control
Ts65Dn
DS patients
NKCC1
Tubulin
*
200
NKCC1/actin (% of Control)
NKCC1/actin (% of WT)
200
150
100
50
0
WT
Ts65Dn
*
100
0
Control
DS
Brain and Tissue Bank for Developmental Disorders
University of Maryland
In collaboration with Dr. Vicari, Bambino Gesu’, Rome, Italy
Drug Development is a High-Risk Investment
for Time and Costs
Chemistry/Pharmacology
Pharmacology
Tests on different
animal species
Healthy
volunteers
$450 million
Patients
Patients
$700 million
$800 million
A rough estimate:
2 million euros
(150 patient, phase 2)
SNC Drug Development
is Even More Risky
In SNC Drug Development
the Outcome is Rather Subjective
Paucity of biomarkers
Based on feelings
Easier placebo effect (patients & families)
A lot is unknown and SNC is difficult to access
Drug Repositioning
May Reduce the Risks of Drug Development
Drug Development vs Repositioning
Chemistry/Pharmacology
Pharmacology
Tests on different
animal species
Healthy
volunteers
Patients
Patients
12 years
$200K
$450
million
Pharma
Company
Gabriele, Martina, Shovan
$700 million
$800 million
Drug Repositioning: the Dark Side
• Side Effects
• Intellectual propriety issues
• Commercial issues
New-Drug Development
after Repositioning May have Lower Risks
Chemistry/Pharmacology
Pharmacology
Tests on different
animal species
Healthy
volunteers
$450 million
Patients
Patients
$700 million
$800 million
Conclusions
Development
Proliferation
Differentiation
Migration
Drug treatment
Repositioning
vs
New Entities
Diseased Brain
Healthy Brain
Take Home Message
The study of brain development by the
new technological tools is a
fundamental step to understand
neurodevelopmental disorders and
design future therapies
Take Home Message
Basic scientists and industry can work
together to discover
new therapeutic approaches
What Can You Do for Us?
Raise awareness without false hopes
We will get the best students
Shovan
Gabriele
Andy
Joanna
Ignacio
Laura
You will guarantee support to Foundations
Acknowledgements
IIT:
EXTERNAL:
Lab: Alessandro Alabastri, Guillaume Bony,
Andrzej Cwetsch, Gabriele Deidda, Ignacio
Fernandez, Francesco Gentile, Shovan Naskar,
Laura Perlini, Bruno Pinto, Annalisa Savardi,
Joanna Szczurkowska
NBT: Marco dal Maschio, Diego Ghezzi, Marco
Canossa, Emanuela Zuccaro, Beatrice Vignoli,
Andrea Contestabile, Martina Parrini, Francesco
Papaleo, Francesca Managò, Fabio Benfenati,
Valter Tucci, Celina Garcia, Giacomo Pruzzo
Nanostructure s Dep: Alessandro Alabastri,
Remo Proietti Zaccaria, Enzo Di Fabrizio
IIT-NEST: Gian Michele Ratto
Maya Shelly (Stony Brook, University)
Mu-ming Poo (UC Berkeley)
Bryan Ballif (University of Vermont)
Yehezkel Ben-Ari (INMED, Marseille)
Jean-Bernard Manent (INMED, Marseille)
Evelyne Sernagor (NewCastle University)
Graziella di Cristo (University of Montreal)
Shubha Tole (Tata Institute, Mumbai)
Fernando Montani (University of Buenos Aires)
Michael Schaefer (Univ. of Maintz,Germany)
Matteo Caleo, Manuela Allegra (CNR, Pisa)
Yuri Bozzi, Giulia Zunino (University of Trento)
Giorgio Carmignoto (CNR, Padova)
Gian Michele Ratto (NEST, Pisa)
Funding:
Interdisciplinary/interdepartmental
grant program