Yr Athro Riccardo Brambilla

Cyhoeddiad

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Ymchwil

I have always been fascinated by molecules and in particular by their function in the nervous system. The whole field of neuropsychopharmacology deals with “drugs” that bind specific target molecules in brain cells and by doing so affect behaviour. In the last few decades, novel powerful genetic technologies have been introduced in the field of neuroscience allowing to modify the expression of specific genes and proteins in a tissue and cell specific manner, thus providing unprecedented novel tools to study the brain. I pioneered the use of “gene knockout” and “viral mediated transgenesis” in the field of learning and memory and cognitive disorders. More recently, my laboratory has designed and validated a number cell penetrating peptides (CPPs) that have the capability to enter the brain and disrupt protein-protein interactions, formidable pharmacological and reversible tools not only to investigate the molecular mechanisms of cognitive deficits but also to set the basis for innovative therapies in patients.

Role of Ras-ERK signalling in memory

Learning and memory processes require that the immediate synaptic information generated by neurotransmitter release is integrated at the cellular and network levels. An essential set of intracellular mechanisms leading to long-term, protein synthesis dependent memories, involved protein kinase cascades. ERK is a major intracellular pathway both controlling cytoplasmic events such as local protein translation and gene expression/chromatin remodelling in the nucleus. The ERK cascade is activated by the Ras family of small GTPases, which before the 1990s were believed to be exclusively involved in cell proliferation and cell survival. In 1992, as a PhD student, I contributed to the cloning of Ras-GRF1, the first ever identified guanine nucleotide exchange factor for Ras, which is essential to activate these small GTPases in response to external stimuli. To our surprise, Ras-GRF1 is exclusively expressed in post-mitotic neurons of the central nervous system, suggesting a specific role of Ras-ERK signalling in adult brain functions. In 1997, as a postdoc at the EMBL in Heidelberg, I generated the Ras-GRF1 KO mouse strain that was the first published genetic model demonstrating a direct involvement of the Ras-ERK signaling cascade in behavioural plasticity (Brambilla et al, Nature, 1997, cited so far 379 times).  Subsequently, as a PI in Milan, I published another important paper demonstrating that ERK1 kinase is a negative regulator of global ERK signaling (Mazzucchelli et al, 2002, Neuron, cited so far 367 times). That paper led us to suggest that the ERK1/ERK2 protein ratio is a major predictive indicator of behavioural and synaptic plasticity changes, particularly in the striatum. The model has been subsequently validated in vitro by my laboratory and has received a number of in vivo confirmations (Vantaggiato et al, 2006, J Biol, cited so far 168 times), supporting the notion that in the absence of ERK1, the remaining ERK2 kinase is facilitated in its nuclear translocation (Marchi et al, PLOSone 2008). Importantly, this model is very essential to explain the phenotypes observed both in 16p11.2 deletion and duplication mouse models, where ERK1 gene is present in one or three copies, respectively. Also, the model has led the development of a novel pharmacological tool, an ERK selective positive modulator.

-R. Brambilla, et al, and  R. Klein. A role for the Ras signaling pathway in synaptic transmission and long-term memory. Nature (1997) 390, 281-286.

-C. Mazzucchelli, et al, and R. Brambilla. Knockout of ERK1 MAP kinase enhances synaptic plasticity in the striatum and facilitates striatal-mediated learning and memory. Neuron (2002) 34, 807-820.

-C. Vantaggiato , I. et al, and R. Brambilla. ERK1 and ERK2 mitogen-activated protein kinases affect Ras-dependent cell signaling differentially. J. Biol. (now BMC Biology)(2006), 5:14.

-M. Marchi, A. D’Antoni, I. Formentini, R. Parra, R. Brambilla, G. M. Ratto and M. Costa. The N-terminal non catalytic domain of ERK1 MAP kinase is responsible for the functional differences with ERK2. PLOS ONE. 2008;3(12):e3873. Epub 2008 Dec 4

Ras-ERK signalling in striatal-dependent plasticity disorders

In the last decade my lab has contributed to expand our knowledge of the role of Ras-ERK signalling in behavioural plasticity. More specifically, I have published a number of important papers involving this pathway in striatal dependent synaptic plasticity and in the responses to drugs of abuse (Fasano et al, 2009; Papale et al, 2016, E-Life). In addition, I have shown that ERK signalling is an important mediator the abnormal involuntary movements caused by L-DOPA, the gold standard therapy for Parkinson’s Disease (Fasano et al, 2010, PNAS; Cerovic et al, 2015 Biol Psych).

-S. Fasano, et al, and R. Brambilla. Ras-Guanine Nucleotide-Releasing Factor 1 (Ras-GRF1) Controls Activation of Extracellular Signal-Regulated Kinase (ERK) Signaling in the Striatum and Long-Term Behavioral Responses to Cocaine. Biol Psychiatry, 66:758 –768 (2009). Doi:10.1016/j.biopsych.2009.03.014

-Fasano, S., et al,  and Brambilla R. Inhibition of Ras-GRF1 in the striatum reverts motor symptoms associated to L-DOPA induced Dyskinesia. PNAS 107, 21824–21829 (2010). Doi:10.1073/pnas.1012071107

-Cerovic M, et al, Brambilla R. Derangement of Ras-Guanine Nucleotide-Releasing Factor 1 (Ras-GRF1) and Extracellular Signal-Regulated Kinase (ERK) Dependent Striatal Plasticity in L-DOPA-Induced Dyskinesia. Biol Psych (2015). 10.1016/j.biopsych.2014.04.002.

-Papale A*, et al, Brambilla R#, and Fasano S. Impairment of cocaine-mediated behaviors by clinically relevant Ras-ERK inhibitors. eLife 2016;5:e17111. DOI: 10.7554/eLife.17111. # co-corresponding author

Cell signalling in Intellectual Disability and neurodegenerative disorders: search for effective treatments

In recent year my laboratory has been involved in a number of projects with the aim of testing novel therapeutic approaches for cognitive disorders in which ERK signaling is deregulated. In a work published in 2017, we demonstrated that in a mouse model of a severe form of Rasopathies, the K-Ras G12V model, aberrant GABAergic synaptogenesis mediated by ERK signalling can be rescued by administration during postnatal development of cell penetrating peptides we devised to attenuate ERK signalling (Papale et al, 2016; Papale et al, 2017). With a similar approach, we also showed in a mouse model of 16p11.2 deletion, that both brain anatomical and behavioural deficits can be rescued (Pucilowska et al, 2018). This last publication support part of the work in the present grant application. More recently, our work using the novel ERK positive modulator indicated that both neurodegeneration and cognitive impairments in mouse models of Alzheimer’s and Huntington’s Disease can be prevented by stimulating this signalling pathway. In the recently awarded MRC Programme Grant, we are planning to use a similar approach to treat a mouse model of 16p11.2 duplication, in which our preliminary data indicate a significant reduction of ERK activity in the brain (Indrigo et al, 2017; Indrigo et al, 2018).

-Papale A, et al, and Brambilla R. Severe intellectual disability and enhanced GABAergic synaptogenesis in a novel model of rare RASopathies. Biol Psych (2017). doi: 10.1016/j.biopsych.2016.06.016.

-J. Pucilowska, et al, R. Brambilla and G.E. Landreth. Pharmacological inhibition of the ERK Signaling Pathway Rescues Cellular and Behavioural impairments associated with 16p11.2 Chromosomal Deletion in Mice. J Neurosci. 2018 Jun 22. pii: 0515-17. doi: 10.1523/JNEUROSCI.0515-17.2018.

-Indrigo I, et al, and Riccardo Brambilla# and Stefania Fasano. Modulation of ERK1/MAPK3 potentiates ERK nuclear signalling, facilitates neuronal cell survival and improves memory in mouse models of neurodegenerative disorders. bioRxiv (2018). doi: https://doi.org/10.1101/496141. # co-corresponding author

- M. Indrigo, A. Papale, S. Fasano and R. Brambilla. Neuroprotective peptide (GB1719520.7, November 27, 2017)(PCT/GB2018/053384, November 23, 2018)

Addysgu

Yn ystod yr ychydig flynyddoedd diwethaf ym Mhrifysgol Caerdydd, rwyf wedi bod yn ymwneud ag addysgu israddedig ac ôl-raddedig ac yn ystod blynyddoedd blaenorol ym Milan goruchwyliais 11 o fyfyrwyr PhD a mwy nag 20 o fyfyrwyr meistr.  Fel Athro Niwrowyddoniaeth a Niwroseicoffarmacoleg, rwy'n darlithio'n rheolaidd ar bynciau amrywiol, o ffarmacoleg sylfaenol cyffuriau sy'n effeithio ar yr ymennydd, i fecanweithiau cellog anhwylderau niwroseiciatrig.

Bywgraffiad

I am Professor of Neuroscience at Cardiff University, and Principal Investigator at the Neuroscience and Mental Health Research Institute (NMHRI). I have been working in the field of signal transduction since 1987 and in molecular neuroscience since 1993. In Heidelberg, as post-doctoral fellow in the laboratory of Ruediger Klein at the European Molecular Biology Laboratory (EMBL), I learned mouse genetics techniques such as gene targeting and successfully applied them to the neurobiology of learning and memory. During the last 22 years as a PI I have continued to work on the role of synaptic signalling in behavioural plasticity and neuropsychiatric disorders. The main focus of my lab is to investigate the cell signalling mechanisms of behavioural dysfunction and test innovative therapeutic approaches not only in mouse models of basal ganglia disorders (drug addiction, Parkinson’s and Huntington’s disease), but also of neurodevelopmental conditions such as intellectual disability (ID) and autism spectrum disorder (ASD). In order to achieve this goal, I have pioneered the use of viral vector technologies to manipulate gene expression in vivo in the brain, to rescue behavioural impairments. More recently, I have also developed novel pharmacological approaches through the use of cell penetrating peptides (CPPs), as efficient means to modulate protein-protein interactions. The validation of CPPs modulating ERK signalling has led to several publications demonstrating that this approach is effective in treating mouse models of cocaine-dependent behaviour, ID, ASD and neurodegenerative disorders. As a complementary approach, my lab is also repurposing clinically relevant brain penetrating drugs, and such studies have recently led to the identification of an ERK pathway inhibitor, PD325901, as a potentially valid compound for ERK-dependent brain disorders. Currently, I am PI of a recent major award (1.5m GBP) from the Medical Research Council to develop biomarkers, human iPSCs and novel therapies for 16p11.2 deletion and duplication CNVs, further expanding my research goal into translational neuroscience.

Papale A, Morella IM, et al, Brambilla R#, and Fasano S. Impairment of cocaine-mediated behaviors by clinically relevant Ras-ERK inhibitors. eLife 2016;5:e17111. DOI: 10.7554/eLife.17111. # co-corresponding author

Papale A, et al, and Brambilla R. Severe intellectual disability and enhanced GABAergic synaptogenesis in a novel model of rare RASopathies. Biol Psych (2017). doi: 10.1016/j.biopsych.2016.06.016.

Pucilowska J, et al, Brambilla R and Landreth GE. Pharmacological inhibition of the ERK Signaling Pathway Rescues Cellular and Behavioural impairments associated with 16p11.2 Chromosomal Deletion in Mice. J Neurosci. 2018 Jun 22. pii: 0515-17. doi: 10.1523/JNEUROSCI.0515-17.2018.

Indrigo M, et al, Brambilla R# and Fasano S. Modulation of ERK1/MAPK3 potentiates ERK nuclear signalling, facilitates neuronal cell survival and improves memory in mouse models of neurodegenerative disorders. bioRxiv (2018). doi: https://doi.org/10.1101/496141. # co-corresponding author

Morè L, Lauterborn JC, Papaleo F and Brambilla R. Enhancing Cognition through Pharmacological and Environmental Interventions: Examples from Preclinical Models of Neurodevelopmental Disorders. Neuroscience and Biobehavioral Reviews (Invited Review). Neurosci Biobehav Rev. 2019 Apr 10. pii: S0149-7634(18)30292-6. doi: 10.1016/j.neubiorev.2019.02.003.

Bibliometric Figures

I have so far published 70 peer-reviewed papers, with a mean citation per publication of 71, 22% of papers cited > 100 times, and H index=34 (Source: Scopus).

Web references:

https://orcid.org/0000-0003-3569-5706

https://www.scopus.com/authid/detail.uri?authorId=7005207779

Education

University of Milan (Milan, Italy) BSc 1988

University of Milan (Milan, Italy) PhD 1992

European Molecular Biology Laboratory (Heidelberg, Germany) Post-Doc 1992-1997

Academic Career and Research Experience

February 1988: Degree in Biological Sciences (110/110 cum laude), University of Milan, Italy. Title of the experimental thesis: "Dissociation of the ligand and dephosphorylation of the receptor for Platelet Derived Growth Factor". Supervisor: Prof. Emmapaola Sturani.

May 1992:  Final examination and awarding of Ph.D in Molecular and Cellular Biology, University of Milan, Italy. Title of the Ph.D. thesis: "Molecular cloning of a putative regulator of mammalian Ras proteins, functionally and structurally homologous to the CDC25 gene product of Saccharomyces cerevisiae". Supervisor: Prof. Emmapaola Sturani.

January 1992- June 1993: Postdoctoral fellow in Giulio Draetta's Laboratory at the European Molecular Biology Laboratory, Heidelberg, Germany.

July 1993- December 1997: Postdoctoral fellow in Ruediger Klein’s Laboratory at the European Molecular Biology Laboratory, Heidelberg, Germany

January 1998- December 2002: Junior Group Leader and FIRC fellow at the San Raffaele Scientific Institute, Milano, Italy.

2003-2015: Senior Group Leader, Research Unit Head (Molecular Genetics of Behaviour), San Raffaele Scientific Institute, Milano, Italy.

2004-2015: Reader in Neuroscience, School of Biosciences, Cardiff University, Cardiff, UK

2013-2016: Visiting Scientist, Istituto di Ricerche Farmacologiche Mario Negri, Milano, Italy.

2017: Visiting Professor, Katholieke Universiteit Leuven, Belgium

2017: Visiting Professor, University of Cagliari, Italy

June 2015-present: Professor of Neuroscience and Neuropsychopharmacology, Cardiff School of Biosciences, Cardiff University, UK.

Anrhydeddau a dyfarniadau

1988: Cymrodoriaeth, Rhaglen PhD mewn Bioleg Cellog a Moleciwlaidd, Prifysgol Milano

1992: Gwobr Cymrodoriaeth Deutscher Akademischer Austauschdienst

1993: Gwobr Cymrodoriaeth Rhaglen Hyfforddiant Cymunedol a Symudedd Ymchwilwyr Ewropeaidd

1994: Gwobr gymrodoriaeth Rhaglen Gwyddoniaeth Frontier Dynol (HFSP)

1996: Gwobr Rhaglen Biotechnoleg Gymunedol Ewrop

1998-2002: Ysgoloriaeth Ffederasiwn Ymchwil Canser yr Eidal (FIRC)

1998: Cymrodoriaeth Tymor Byr HFSP

1999: Gwobr 10fed Pen-blwydd Rhaglen Gwyddoniaeth Frontier Dynol

2001: Cymrodoriaeth tymor byr Sefydliad Bioleg Moleciwlaidd Ewropeaidd (EMBO)

Ers 1997: Aelod o'r Gymdeithas Niwrowyddoniaeth (SfN)

Ers 2002: Aelod o Gymdeithas Niwrowyddoniaeth yr Eidal (SINS)

Pwyllgorau ac adolygu

2014-presennol: Aelod Panel Grant Cydgrynhoi LS5 "Niwrowyddorau ac Anhwylderau Niwrolegol" y Cyngor Ymchwil Ewropeaidd (ERC)

2002-presennol: Aelod o'r Gymdeithas Gwybyddiaeth Moleciwlaidd a Cellog (MCCS, Cymrawd y Cyngor Sefydlu)

2006-2012: Llywydd Sefydlu'r MCCS Ewropeaidd (EMCCS)

1998-presennol: Adolygydd ar gyfer Asiantaethau Grantiau gan gynnwys Rhaglen NEST a FP6/FP7 y Comisiwn Ewropeaidd, Ffrainc, Sbaeneg, Ffleminaidd a Gweinyddiaethau Ymchwil Iseldiroedd, y MRC Prydeinig, Sefydliad Gwyddoniaeth Dwywladol yr Unol Daleithiau-Israel, Sefydliad Gwyddoniaeth Israel Israel, Sefydliad Gwyddoniaeth Israel, Parkinson's UK

2008-2009: Llywydd ac Aelod o AERES (Agence d'Evalutation de la Recherche et de l'Enseignement Superieur, Ffrainc) Paneli (Niwrowyddoniaeth)

2011-2014: Aelod Panel ANR (Agence Nationale de la Recherche, Ffrainc) (Niwrowyddoniaeth a Ffisiolegol)

2012-presennol: Aelod o Bwyllgor Gweithredol MCCS Ewrop (EMCCS)

1998 - presennol: Adolygydd Cyfnodolion Niwrowyddoniaeth gan gynnwys Neuron, Journal of Neuroscience, Moleciwlaidd a Cellog Niwrowyddoniaeth, Ymchwil i'r Ymennydd, TINS, Niwrobioleg Clefydau, Seiciatreg Fiolegol, Journal of Neuroscience Methods, Journal of Neurochemistry, Neuroscience, PlosONE, Dysgu a Chof, Geneteg Moleciwlaidd Dynol.

2007-2010: adolygiad Golygydd Frontiers mewn Niwrowyddoniaeth Ymddygiadol

2010-presennol: Golygydd Cysylltiedig, Ffiniau mewn Niwrowyddoniaeth Ymddygiadol

2011-presennol: Aelod o Fwrdd Golygyddol Synapse