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