News

Reconstitution of the Human Nigro-striatal Pathway on-a-Chip Reveals OPA1-Dependent Mitochondrial Defects and Loss of Dopaminergic Synapses

Stem cell-derived neurons are generally obtained in mass cultures that lack both spatial organization and any meaningful connectivity. We implement a microfluidic system for long-term culture of human neurons with patterned projections and synaptic terminals. Co-culture of human midbrain dopaminergic and striatal medium spiny neurons on the microchip establishes an orchestrated nigro-striatal circuitry with functional dopaminergic synapses. We use this platform to dissect the mitochondrial dysfunctions associated with a genetic form of Parkinson’s disease (PD) with OPA1 mutations. Remarkably, we find that axons of OPA1 mutant dopaminergic neurons exhibit a significant reduction of mitochondrial mass. This defect causes a significant loss of dopaminergic synapses, which worsens in long-term cultures. Therefore, PD-associated depletion of mitochondria at synapses might precede loss of neuronal connectivity and neurodegeneration. In vitro reconstitution of human circuitries by microfluidic technology offers a powerful system to study brain networks by establishing ordered neuronal compartments and correct synapse identity.

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News

Whole brain delivery of an instability-prone Mecp2 transgene improves behavioral and molecular pathological defects in mouse models of Rett syndrome

Rett syndrome (RTT) is an incurable neurodevelopmental disorder caused by mutations in the gene encoding for methyl-CpG binding-protein 2 (MeCP2). Gene therapy for this disease presents inherent hurdles since MECP2 is expressed throughout the brain and its duplication leads to severe neurological conditions as well. However, the recent introduction of AAV- PHP.eB, an engineered capsid with an unprecedented efficiency in crossing the blood-brain barrier upon intravenous injection, has provided an invaluable vehicle for gene transfer in the mouse nervous system. Herein, we use AAV-PHP.eB to deliver an instability-prone Mecp2 (iMecp2) transgene cassette which, increasing RNA destabilization and inefficient protein translation of the viral Mecp2 transgene, limits supraphysiological Mecp2 protein levels in transduced neural tissues. Intravenous injections of the PHP.eB-iMecp2 virus in symptomatic male and female Mecp2 mutant mice significantly ameliorated the disease progression with improved locomotor activity, coordination, lifespan and normalization of altered gene expression and mTOR signaling. Remarkably, PHP.eB-iMecp2 administration did not result in severe toxicity effects either in female Mecp2 mutant or in wild-type animals. In contrast, we observed a strong immune response to the transgene in treated male Mecp2 mutant mice that was overcome by immunosuppression. Overall, PHP.eB-mediated delivery of the iMecp2 cassette provided widespread and efficient gene transfer maintaining physiological Mecp2 protein levels in the brain. This combination defines a novel viral system with significant therapeutic efficacy and increased safety which can contribute to overcome the hurdles that are delaying clinical applications of gene therapy for RTT.

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Individuato un gene implicato in molte disabilità intellettive

La scoperta viene da un gruppo di ricerca dell’Ospedale San Raffaele e del Cnr e potrebbe rappresentare il primo passo per mettere a punto terapie per diversi disturbi

Dalle difficoltà di movimento e linguaggio ai comportamenti tipici dello spettro autistico: la responsabilità di tutto questo sarebbe imputabile al malfunzionamento di un gene chiamato Setd5. La scoperta , che apre nuove prospettive nel trattamento di questi disturbi, viene da un gruppo di ricercatori dell’Irccs Ospedale San Raffaele di Milano e dell’Istituto di Neuroscienze del Consiglio Nazionale delle Ricerche (Cnr-In) che hanno condotto uno studio pubblicato sulla prestigiosa rivista internazionale Neuron.

Sotto il nome di disturbi del neurosviluppo va un insieme di patologie neurologiche e psichiatriche che di solito si manifestano durante l’età dello sviluppo, condizioni complesse e molto diversificate tra loro, causate da un insieme di fattori sia genetici sia ambientali. I risultati dello studio condotto dai ricercatori coordinati da Alessandro Sessa e Vania Broccoli, in collaborazione con i gruppi delle università di Trento e Pisa (coordinati rispettivamente da Alessio Zippo e Massimiliano Andreazzoli) dimostrano che SETD5 codifica per una proteina con un ruolo fondamentale e inatteso: quello di assicurare la corretta trascrizione delle informazioni del Dna.

Meccanismo genetico

Fino a oggi non era chiara quale fosse l’esatta funzione di SETD5, né il meccanismo per il quale, a fronte di mutazioni che mettono il gene fuori gioco, si manifestino problemi nello sviluppo del sistema nervoso. «SETD5 è una specie di “architetto molecolare” che regola la complessa organizzazione del Dna dei neuroni del nostro cervello. Quando è fuori gioco, le informazioni contenute nel Dna vengono trascritte in maniera alterata ed incompleta», spiega Alessandro Sessa, coordinatore della ricerca e primo autore dello studio. «I topi privi di questo gene mostrano comportamenti anomali sia dal punto di vista cognitivo che sociale». Nonostante le mutazioni nel gene SETD5 siano implicate solo in una porzione dei casi disabilità intellettive, i meccanismi individuati potrebbero essere comuni anche ad altri geni che svolgono funzionalità simili, oltre a suggerire nuove ipotesi di studio. La ricerca è stata finanziata dal Ministero della Salute e da Fondazione Telethon.

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Un gene mutato alla base di alcune disabilità intellettive

Gruppo di ricercatori italiani scopre il suo meccanismo d'azione

Identificato il meccanismo molecolare all'origine di alcune forme di disabilità intellettive, che colpiscono linguaggio e movimento, spesso accompagnate da manifestazioni autistiche: è collegato alla mutazione di un gene (Setd5). La scoperta, descritta sulla rivista Neuron, si deve ad un gruppo di ricercatori italiani, coordinati dall'Ospedale San Raffaele di Milano e l'Istituto di Neuroscienze del Consiglio Nazionale delle Ricerche (Cnr-In), in collaborazione con le università di Trento e Pisa.

Le disabilità intellettive possono essere associate alle mutazioni di moltissimi geni diversi. Tra questi il gene Setd5, noto da tempo, ma di cui non si conosceva l'esatta funzione né il meccanismo per cui si manifestano problemi nello sviluppo del sistema nervoso. Setd5 è una sorta di 'architetto molecolare', che regola l'organizzazione del Dna dei neuroni del cervello.

"Quando è fuori gioco, le informazioni nel Dna vengono trascritte in modo alterato e incompleto", spiega Alessandro Sessa del San Raffalele, coordinatore della ricerca. "Ciò influisce sulla formazione e il corretto funzionamento dei neuroni. I topi privi di questo gene mostrano infatti comportamenti anomali sia dal punto di vista cognitivo che sociale".

Anche se le mutazioni di questo gene sono la causa genetica solo di una parte dei casi disabilità intellettive e autismo, i meccanismi individuati potrebbero essere comuni ad altri geni che svolgono funzioni simili. "Conoscere il meccanismo molecolare alla base di una patologia - conclude Sessa - è il primo passo per individuare dei possibili bersagli terapeutici".

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Identificato il meccanismo molecolare all’origine di alcune forme di disabilità intellettive

Gruppo di ricercatori italiani scopre il suo meccanismo d'azione

Un team di ricerca dell’Irccs Ospedale San Raffaele di Milano e dell’Istituto di neuroscienze del Consiglio nazionale delle ricerche (Cnr-In) ha scoperto il meccanismo di azione di un gene (Setd5) la cui mutazione è associata ad alcune forme di disabilità intellettive, spesso accompagnate da manifestazioni autistiche. Lo studio, svolto in collaborazione con le università di Trento e Pisa, è pubblicato su "Neuron"

I disturbi del neurosviluppo sono un insieme ampio di patologie neurologiche e psichiatriche che si manifestano durante l’età dello sviluppo e di cui fanno parte le disabilità intellettive e i disturbi nello spettro autistico. Si tratta di condizioni complesse e molto diversificate tra loro, causate da un insieme di fattori sia genetici sia ambientali.

Un gruppo di ricercatori dell’Irccs Ospedale San Raffaele di Milano e dell’Istituto di neuroscienze del Consiglio nazionale delle ricerche (Cnr-In) coordinati dai dottori Alessandro Sessa e Vania Broccoli, in collaborazione con i gruppi delle università di Trento e Pisa – coordinati rispettivamente dal dottor Alessio Zippo e dal professor Massimiliano Andreazzoli – ha scoperto il meccanismo di azione di un gene (Setd5) la cui mutazione è associata ad alcune forme di disabilità intellettive spesso accompagnate da manifestazioni autistiche. I risultati dello studio pubblicato oggi sulla prestigiosa rivista internazionale Neuron, dimostrano che Setd5 codifica per una proteina con un ruolo fondamentale e inatteso all’interno del nucleo dei neuroni: quello di assicurare la corretta trascrizione delle informazioni del Dna. La scoperta apre nuove prospettive nello studio di questi disturbi e getta le basi per l’identificazione di futuri bersagli terapeutici.

Le disabilità intellettive, come molti altri disturbi del neurosviluppo, sono condizioni altamente complesse: grazie al lavoro di gruppi di ricerca di tutto il mondo, oggi sappiamo che possono essere associate a mutazioni di moltissimi geni diversi. Tra questi c’è un gene chiamato Setd5, che quando non più funzionante dà origine a forme di disabilità intellettiva che colpiscono in particolare il linguaggio e il movimento, oltre a comportamenti tipici dello spettro ossessivo-compulsivo e dello spettro autistico. Sebbene questo sia noto da tempo, fino a oggi non era chiara quale fosse l’esatta funzione di Setd5, né il meccanismo per il quale, a fronte di mutazioni che mettono il gene fuori gioco, si manifestino problemi nello sviluppo del sistema nervoso.

Attraverso lo studio di sistemi cellulari e modelli animali della mutazione, i ricercatori hanno scoperto che Setd5 gioca un ruolo chiave per la struttura del Dna e quindi per il corretto recupero delle informazioni in esso presenti. Queste informazioni infatti, per poter essere utilizzate, devono prima essere trascritte in un'altra forma – l’Rna, una sorta di copia carbone del Dna – e portate fuori dal nucleo. Setd5 è coinvolto proprio nel regolare questo delicato processo di trascrizione delle informazioni.

“Setd5 rappresenta una sorta di ‘architetto molecolare’ che regola la complessa organizzazione del Dna dei neuroni del nostro cervello. Quando è fuori gioco, le informazioni contenute nel Dna vengono trascritte in maniera alterata ed incompleta”, spiega Alessandro Sessa, coordinatore della ricerca e primo autore dello studio. “Secondo quando abbiamo osservato, questo impatta in particolare sulla formazione e sul corretto funzionamento dei neuroni. Come conseguenza i topi privi del gene Setd5 mostrano comportamenti anomali sia dal punto di vista cognitivo che sociale”.

Nonostante le mutazioni nel gene Setd5 rappresentino la causa genetica solo di una porzione dei casi disabilità intellettive e autismo, i meccanismi individuati potrebbero essere comuni anche ad altri geni che svolgono funzionalità simili, oltre a suggerire nuove ipotesi di ricerca: “Conoscere il meccanismo molecolare alla base di una patologia è il primo passo per individuare dei possibili bersagli terapeutici”, conclude Sessa.

La ricerca è stata finanziata dal Ministero della Salute e da Fondazione Telethon.

Link

Research

The human brain is made of 100 billion neurons (and many more glial cells), each of them making at least 1,000 connections with other neurons for a total of 100 trillion connections with distinct and peculiar activities. Despite this breathtaking complexity, new results have provided a solid proof-of-concept that cell and gene therapy approaches have the outstanding potential to replace neurons, reconstitute brain circuits, and repair synaptic dysfunctions in the adult brain up to block or revert brain damages or diseases.

Our mission is to develop new therapeutics for neurological disorders waiting for an effective, safe and long-lasting treatment. The significant step-forward in our understanding of the molecular pathophysiological roots of these diseases has disclosed unprecedented opportunities for establishing new therapeutic strategies for these serious illnesses. Remarkably, the advent of novel and powerful technologies such as stem cell reprogramming, systems biology, gene editing approaches (TALENs and CRISPR/Cas9) and gene therapy vectors has provided an invaluable and powerful tool-box to model, understand and treat these disorders. We are taking advantage of these technologies to implement new pathways of medicine discovery and set innovative therapeutic strategies to cure diseases. Over the years, we have developed a strong commitment to exploit therapeutics for incurable infantile neurological disorders with cognitive dysfunctions and untreatable epilepsy. Additionally, our efforts are aimed to establish new cures for Parkinson’s disease and MSA causing dopaminergic cell loss and mental impairment.

Stay tune on this website to follow our developments and news in seeking for new discoveries and therapeutics!

Ideas are cheap; models are cheap; experiments are golden.

Scientists have long been known to put too much faith in their model and not enough faith in their data.

Believe your data, not the model that is popular with you or with the community.

All models will be wrong sooner or later.

Chris Walsh

Ongoing Research Initiatives

Epigenetic mechanisms of brain development

and neuroinfantile disorders

Coming soon

iPS cell technology for

in vitro disease modeling

Coming soon

Developing gene therapy approaches

for neurological disorders

Coming soon

Pathological mechanisms of neuronal cell

loss in Parkinson’s disease

Coming soon

Genetic strategies of

neuronal cell reprogramming

Coming soon

Novel therapies based on

gene editing technologies

Coming soon

Novel translational research

programs for Dravet syndrome

Coming soon

New therapies for

Parkinson’s disease

Coming soon

Selected Publications

Pharmacological Inhibition of Necroptosis Protects from Dopaminergic Neuronal Cell Death in Parkinson’s Disease Models

Iannielli A, Bido S, Folladori L, Segnali A, Cancellieri C, Maresca A, Massimino L, Rubio A, Morabito G, Caporali L, Tagliavini F, Musumeci O, Gregato G, Bezard E, Carelli V, Tiranti V, Broccoli V

Cell Reports, 22(8), 2066–2079. doi: 10.1016/j.celrep.2018.01.089

TBR2 antagonizes retinoic acid dependent neuronal differentiation by repressing ZFP423 during corticogenesis

Massimino L, Flores-Garcia L, Di Stefano B, Colasante G, Icoresi-Mazzeo C, Zaghi M, Hamilton BA, Sessa, A

Developmental Biology, 434(2), 231–248. doi: 10.1016/j.ydbio.2017.12.020

Cas9/sgRNA selective targeting of the P23H Rhodopsin mutant allele for treating Retinitis Pigmentosa by intravitreal AAV9.PHP.B-based delivery

Giannelli SG, Luoni M, Castoldi V, Massimino L, Cabassi T, Angeloni D, Demontis G, Leocani L, Andreazzoli M, Broccoli V.

Hum Mol Genet. 2017 Dec 21. doi: 10.1093/hmg/ddx438

The Tbr2 Molecular Network Controls Cortical Neuronal Differentiation Through Complementary Genetic and Epigenetic Pathways

Sessa A, Ciabatti E, Drechsel D, Massimino L, Colasante G, Giannelli S, Satoh T, Akira S, Guillemot F, Broccoli V.

Cereb Cortex. 2017 Jun 6;27(12):5715. doi: 10.1093/cercor/bhw270

A Human Bi-specific Antibody against Zika Virus with High Therapeutic Potential

Wang J, Bardelli M, Espinosa DA, Pedotti M, Ng TS, Bianchi S, Simonelli L, Lim EXY, Foglierini M, Zatta F, Jaconi S, Beltramello M, Cameroni E, Fibriansah G, Shi J, Barca T, Pagani I, Rubio A, Broccoli V, Vicenzi E, Graham V, Pullan S, Dowall S, Hewson R, Jurt S, Zerbe O, Stettler K, Lanzavecchia A, Sallusto F, Cavalli A, Harris E, Lok SM, Varani L, Corti D.

Cell. 2017 Sep 21;171(1):229-241.e15. doi: 10.1016/j.cell.2017.09.002

AAV-PHP.B-Mediated Global-Scale Expression in the Mouse Nervous System Enables GBA1 Gene Therapy for Wide Protection from Synucleinopathy

Morabito G, Giannelli SG, Ordazzo G, Bido S, Castoldi V, Indrigo M, Cabassi T, Cattaneo S, Luoni M, Cancellieri C, Sessa A, Bacigaluppi M, Taverna S, Leocani L, Lanciego JL, Broccoli V.

Mol Ther. 2017 Dec 6;25(12):2727-2742. doi: 10.1016/j.ymthe.2017.08.004

Reprogramming of somatic cells: iPS and iN cells

Broccoli V.

Prog Brain Res. 2017;230:53-68. doi: 10.1016/bs.pbr.2016.12.009

Two factor-based reprogramming of rodent and human fibroblasts into Schwann cells

Mazzara PG, Massimino L, Pellegatta M, Ronchi G, Ricca A, Iannielli A, Giannelli SG, Cursi M, Cancellieri C, Sessa A, Del Carro U, Quattrini A, Geuna S, Gritti A, Taveggia C, Broccoli V.

Nat Commun. 2017 Feb 7;8:14088. doi: 10.1038/ncomms14088

Rapid and efficient CRISPR/Cas9 gene inactivation in human neurons during human pluripotent stem cell differentiation and direct reprogramming

Rubio A, Luoni M, Giannelli SG, Radice I, Iannielli A, Cancellieri C, Di Berardino C, Regalia G, Lazzari G, Menegon A, Taverna S, Broccoli V.

Sci Rep. 2016 Nov 18;6:37540. doi: 10.1038/srep37540

MyT1 Counteracts the Neural Progenitor Program to Promote Vertebrate Neurogenesis

Vasconcelos FF, Sessa A, Laranjeira C, Raposo AASF, Teixeira V, Hagey DW, Tomaz DM, Muhr J, Broccoli V, Castro DS.

Cell Rep. 2016 Oct 4;17(2):469-483. doi: 10.1016/j.celrep.2016.09.024

Coenzyme A corrects pathological defects in human neurons of PANK2-associated neurodegeneration

Orellana DI, Santambrogio P, Rubio A, Yekhlef L, Cancellieri C, Dusi S, Giannelli SG, Venco P, Mazzara PG, Cozzi A, Ferrari M, Garavaglia B, Taverna S, Tiranti V, Broccoli V, Levi S.

EMBO Mol Med. 2016 Oct 4;8(10):1197-1211. doi: 10.15252/emmm.201606391

Rapid Conversion of Fibroblasts into Functional Forebrain GABAergic Interneurons by Direct Genetic Reprogramming

Colasante G, Lignani G, Rubio A, Medrihan L, Yekhlef L, Sessa A, Massimino L, Giannelli SG, Sacchetti S, Caiazzo M, Leo D, Alexopoulou D, Dell'Anno MT, Ciabatti E, Orlando M, Studer M, Dahl A, Gainetdinov RR, Taverna S, Benfenati F, Broccoli V.

Cell Stem Cell. 2015 Dec 3;17(6):719-734. doi: 10.1016/j.stem.2015.09.002

Histone modifications controlling native and induced neural stem cell identity

Broccoli V, Colasante G, Sessa A, Rubio A.

Curr Opin Genet Dev. 2015 Oct;34:95-101. doi: 10.1016/j.gde.2015.08.003

Direct conversion of fibroblasts into functional astrocytes by defined transcription factors

Caiazzo M, Giannelli S, Valente P, Lignani G, Carissimo A, Sessa A, Colasante G, Bartolomeo R, Massimino L, Ferroni S, Settembre C, Benfenati F, Broccoli V.

Stem Cell Reports. 2015 Jan 13;4(1):25-36. doi: 10.1016/j.stemcr.2014.12.002

Modeling physiological and pathological human neurogenesis in the dish

Broccoli V, Giannelli SG, Mazzara PG.

Front Neurosci. 2014 Jul 24;8:183. doi: 10.3389/fnins.2014.00183

Cerebral Cortex Front Cover

Lab members

Vania Broccoli, PhD

Stem Cells and Neurogenesis Unit
Division of Neuroscience
San Raffaele Scientific Institute
Via Olgettina 58
20132 Milan
broccoli.vania@hsr.it
TEL. Lab +39 02 26434612
TEL. Office +39 02 26434616
FAX +39 02 26436164

Broccoli Vania is leading the Unit of “Stem Cell and Neurogenesis” at the San Raffaele Scientific Institute in Milan (Italy). He is a neurobiologist interested to unravel the molecular mechanisms that control key processes of brain development such as neural stem cell identity maintenance, neural commitment and migration, neural network establishment and function. Lately, his group has applied new technologies of direct cell reprogramming to convert mouse and human skin fibroblasts into functional dopaminergic neurons. Now, he aims to further strengthen these approaches to establish safe and efficient systems for producing functional human neurons suitable for cell replacement therapies in infantile neurological and neurodegenerative disorders. V. Broccoli has published 64 papers in international scientific journals with a Total Impact Factor (IF) = 492.2, with average IF=7.8/Journal, was cited 2732 in total, with an average citation per item of 42.6, H-index is 27 based on the ISI web of science. He contributed 5 chapters to scientific books and edited one special issue of the “Scientific American” Italian Edition.

Vania Broccoli established his laboratory in 2003 at the San Raffaele Scientific Institute and focused on revealing the molecular mechanisms controlling neuronal differentiation and cerebral cortex development. From 2000 to 2001, he worked at the Telethon Institute of Molecular Medicine (TIGEM) as associate scientist. As post-doctoral fellow, from 1996 to 1999, he worked in Prof. W.Wurst’s laboratory in Munich, unraveling the role played by the transcription factor Otx2 in shaping midbrain-hindbrain junction and controlling dopaminergic neuronal ontogenesis. From 1993-1996, as PhD student in Prof. E. Boncinelli’s laboratory, he studied the mechanisms of brain regionalization and differentiation, in particular, focusing on the Emx and Otx transcription factors during cerebral cortex development. He is contract Professor at the Medical School of the Vita-Salute San Raffaele University.

Major Funding of the PI

The Michael J. Fox Foundation ”Assessing the therapeutic potential of iDA neuronal cells in an autologous transplantation strategy. 2012.

Telethon (GGP11110) " Molecular bases and in vitro modeling of Cdkl5 dependent infantile neurological disorders". 2011-2014.

Telethon (GGP09117) "Congenital Rett syndrome: cellular and mouse models for the study of FOXG1 impact on forebrain neurogenesis". 2010-2013.

ERANET-Neuron (EU) "Modeling Parkinson’s disease by iPStechnology: generation of human affected dopaminergic neurons and gene disease correction by site-specific integration" 2009-2012.

Italian Ministry of Health - Young Researcher Award (RF12) “Derivation, characterization and preclinical applications of fibroblasts-derived iPS cells from patients affected by Parkinson’s disease” 2008-2011.

Telethon (GGP07181) “Identification of the Arx molecular mechanisms in controlling telencephalon development and GABAergic neuronal migration: implications for Arx dependent neurological diseases” 2007-2010.

ERANET-Rare Diseases (EU) “EURORETT: European Network onRett Syndrome” 2007-2010.

RSRF Rett Syndrome Research Foundation (USA) "Production and first analysis of an animal model for the “early seizure” Rettsyndrome variant: conditional inactivation of the murine Cdkl5 gene" 2007-2008.

Italian Ministry of Health “Trinucleotide (GCG) repeat expansion of the ARX gene and progressive dystonia in infancy” 2006-2007.

Telethon (GGP04141) "Unravelling the pathogenetic mechanisms of Arx mutations leading to several forms of mental retardation, epilepsy and XLAG" 2004-2006.

Italian National Program for Stem Cell Research (CS71) "Derivation and neural differentiation of mouse and bovine embryonic stem cells: an in vitro and in vivo comparative assessment with neural stem cells." 2004-2005. €120,000

EU, 5th framework program (FP5) "Neural degeneration and control of cell polarity" QLG3-CT-2002-01266. 2000-2002.

Gaia Colasante, PhD

colasante.gaia@hsr.it

Gaia Colasante is a Postdoctoral Fellow in Vania Broccoli’s lab. Currently, she is interested in the generation of functional GABAergic cortical interneurons through direct reprogramming technique, starting from mouse embryonic fibroblasts. To this aim, she is applying her knowledge on cortical interneuron development, gained during her PhD in Neuroscience performed in the same lab and concluded in April 2011. At that time, she studied the role of the transcription factor Arx during telencephalic development, in both GABAergic and glutamatergic compartment.

During her PhD, she had the chance to work in Jeffrey Golden’s laboratory at Children’s Hospital of Philadelphia, establishing a strong and fruitful collaboration.

Serena Giannelli, PhD

giannelli.serena@hsr.it

Serena is a senior Post-doc in Vania Broccoli’s lab. She got her bachelor degree in 2000 in Bologna and then she moved to work in the USA (at the Weill Medical College of Cornell University in New York) to than go back to Italy to join the Broccoli’s lab in late 2005 as a PhD student. During her internship in Molecular Medicine program of Neuroscience at San Raffaele University, she focused on the regenerative potential of the retina and she graduated in 2011. Nowadays she is still engaged in retinal studies but she is also interested in finding new application to exploit the CRISPR/Cas9 system.

Alessandro Sessa, PhD

sessa.alessandro@hsr.it

Alessandro Sessa is a staff scientist in Unit of “Stem Cell and Neurogenesis” at the San Raffaele Scientific Institute in Milan (Italy). He is a developmental neurobiologist interested in the role of epigenetic players in the physiological development of the brain as well as their impact for the establishment of pathological conditions in humans. He obtained his PhD (Molecular Medicine, Neuroscience program at San Raffaele University) in Vania Broccoli’s laboratory in 2012. During his PhD, he worked as guest scientist in Francois Guillemot’s lab at NIMR in London to improve his skills in chromatin immuno-precipitation following by deep sequencing. During his career he combined molecular biology, genetic manipulation and high-throughput analysis to address biological processes during brain formation. Alessandro Sessa has published 20 papers in international scientific peer reviewed journals and contributed to one chapter for the textbook “Epigenetics in Human Disease, 2nd edition” (Elsevier). He is contract Professor at the Medical School of the Vita-Salute San Raffaele University.

Personal Funding:

Italian Ministry of Health: Young Researcher Award - Ricerca Finalizzata Giovani Ricercatori (# GR-2016-02362536); role: PI, collaborator. 2018-2021

Italian Ministry of Health: Young Researcher Award - Ricerca Finalizzata Giovani Ricercatori (# GR-2013-02355540); role: PI, coordinator. 2016-2019

Telethon Foundation: Research Project (# GGP15096); role: PI, coordinator. 2015-2018

Simone Bido, PhD

bido.simone@hsr.it

Thanks to his 10 years of field studies Simone Bido meets the requirements to investigate the mechanism underlying the development of Parkinson’s Disease. He started working in animal models of Parkinson’s disease to investigate the neurochemical changes occurring in the brain structures. The surgery for microdialysis probe implantation, toxins injection and brain dissection gave him the opportunity to improve and refine the skills in the practice of surgery. He mostly worked on the processes underlying the development of L-DOPA-induced dyskinesia, neurochemistry of the movement, mitochondria-mediated neurodegeneration and autophagy.

Simone Brusco, PhD

brusco.simone@hsr.it

My name is Brusco Simone, and I am an electrophysiologist. I completed my Master Degree in Biology at the University of Milano Bicocca under the guidance of Prof. Andrea Becchetti; during my thesis I studied the effect of nitrosamines, tobacco related compound, on the neuronal isoform of nicotinic acetylcholine receptor.

I then completed my PhD in Molecular and Translational Medicine in the same lab, this time focusing on nicotinic receptor mutations involved in Nocturnal Frontal Lobe epilepsy.

During my career I focused on electrophysiology techniques and in particular on patch-clamp.

Currently in Vania’s lab I’m working on Dravet Syndrome, trying to dissect the role of SSRI in the management of this disease and the neuronal defect responsible of this disease.

Camilla Maffezzini, PhD

maffezzini.camilla@hsr.it

Camilla Maffezzini got her MSc degree in Neurobiology at the University of Pavia in 2014. She then moved to Stockholm to start a PhD in Scientific Medicine, that got to an end in 2019: during these years she had the chance to deepen her knowledge of the mitochondrial field and of bioenergetics. Currently, she is interested in the pathogenic mechanisms of Wolfram syndrome.

Sharon Muggeo, PhD

muggeo.sharon@hsr.it

Sharon received her Bachelor degree in Biotechnology (2009) from University of Milan-Bicocca and her Master degree in Molecular and Cellular Medical Biotechology (2011) from Vita-Salute San Raffaele University. She obtained her Ph.D. in Experimental Pathology and Neuropathology in 2015 at Humanitas Research Hospital. She gained expertise in induced pluripotent stem cells (iPSc) and their differentiation to the hematopoietic lineage, in particular to osteoclasts. She studied also the role of the proto-oncogene Pbx1 in myeloproliferative neoplasms. Currently, in Vania Broccoli’s lab, she is applying her knowledge on microglia differentiation from iPSc.

Alicia Rubio Garrido, PhD

rubiogarrido.alicia@hsr.it

Alicia Rubio received her Ph.D. in 2008 at the Centro de Biología Molecular “Severo Ochoa” (Madrid, Spain) in Jesus Avila´s lab. Her work was focused on the study of different signaling pathways implicated in the progression of tau pathology in Alzheimer´s disease. As a Ph.D. student, she had the opportunity to spend 6 months in Luis de Lecea´s lab (Scripps Research Institute and Stanford University, California, US) where she studied more deeply the effect of the cortical neuromodulator cortistatin on tau phosphorylation.

In 2010, she earned a 4-year postdoctoral fellowship to work in the stem cell field in Isabel Fariñas´s lab (University of Valencia, Spain) where she was interested in how the division type of adult neural stem cells can be controlled and modulated. She was also involved in the study of Numb, a fate determinant factor, in the division of stem cells and its role in lineage differentiation in a 2-year collaboration in Pier Paolo di Fiore´s lab (Istituto Europeo Oncologico, Milano, Italy).

In 2014 she joined Vania Broccoli´s lab as a postdoctoral researcher and she currently exploits her interests on neurodegeneration and stem cells differentiation by studying fibroblasts-derived iPS cells from patients with genetic forms of Parkinson´s disease.

Laura Argelich

argelich.laura@hsr.it

Laura Argelich is a PhD student of the International PhD Course in Molecular Medicine at Vita-Salute San Raffaele University (Neuroscience and Experimental Neurology curriculum) since October 2019. She received her Bachelor degree in Biotechnology in 2017 and his Master degree in Biochemistry, Molecular Biology and Biomedicine in 2018. Currently her Ph.D. project is focused on exploiting direct reprogramming strategy to obtain induced dopaminergic neurons to treat parkinsonian mouse models.

Federica Banfi

banfi.federica@hsr.it

Federica Banfi is a Ph.D. student of the International Ph.D. Course in Molecular Medicine at Vita-Salute San Raffaele University (Neuroscience and Experimental Neurology curriculum), under the supervision of Alessandro Sessa, since October 2016. She obtained her Bachelor’s degree in Biological Sciences (2013) and Master’s degree in Molecular Biology of the Cell (2015) at the University of Milan. During her Master internship, in Lorenza Lazzari’s lab at Fondazione IRCCS Ca` Granda Ospedale Maggiore Policlinico Milano, she studied the role of extracellular vesicles derived from human mesenchymal stem cells in intercellular communication. Currently her Ph.D. project is focused on understanding the roles of SETBP1 in brain development and neurodevelopmental diseases.

Angelo Iannielli

iannielli.angelo@hsr.it

Angelo Iannielli is a postgraduate fellow in Vania Broccoli’s lab. He received his Master degree in Molecular Biotechnology from University of Turin in 2013. During his Master internship, he attended research laboratory at Neuroscience Institute Cavalieri Ottolenghi (NICO) where he prepared his master thesis focused on the role of transcriptional factor COUP-TFI in controlling activity-dependent TH expression in adult dopaminergic olfactory bulb interneurons. In August 2014, he joined Vania Broccoli’s lab where he focused on direct reprogramming of human somatic cells into functional induced dopaminergic neurons. Currently, he works on link between mitochondrial dysfunctions and Parkinson’s disease.

Mirko Luoni

luoni.mirko@hsr.it

Mirko Luoni received his degree in Biotechnology (2013) from University of Insubria (Varese) and his master degree in Medical Biotechnology (2016) from Vita-Salute San Raffaele University (Milano). During his Master internship in Broccoli’s Lab, he worked on the application of CRISPR/Cas9 system to develop new methods aiming to quickly analyze the effect of pathological alterations during neuronal differentiation. Currently, he is in the Ph.D program in Neuroscience and Experimental Neurology at San Raffaele University and he works on Rett Syndrome in order to find novel therapeutic strategies.

Gabriele Ordazzo

ordazzo.gabriele@hsr.it

Gabriele Ordazzo received his Bachelor’s degree in Medical and Pharmacological Biotechnology and MD in Cellular and Molecular Biotechnology at Vita-Salute San Raffaele University, Milano. He’s interesting in neuroscience and focus on neurodegeneration field. He’s working on gene therapies and characterization of Parkinson (PD) pathophysiologic mechanisms in cellular and mice model.

Antonio Niro

niro.antonio@hsr.it

Antonio Niro is a Molecular Biology Technician in Vania Broccoli’s Lab. He received his master’s degree in Medical Biotechnology at University of Milano-Bicocca: School of Medicine and Surgery in 2015. After his master’s degree he spent two years in Gambacorti-Passerini’s Molecular Oncology Lab where he studied deeply the identification of novel mutations in aCML patients. He joined the laboratory of Vania Broccoli in 2018 for the project called ‘Generation of genetically modified iPS cell lines using the CRISPR/Cas9 system’. His technical skills cover most of the molecular biology techniques and the CRISPR/Cas9 gene-editing. He also deals with the management of quotes, orders and purchases of laboratory reagents, management of relations with suppliers, maintenance of analytical tools, supervision and general coordination of students in the laboratory.

Laura Pintado

Raffaele Ricci

ricci.raffaele@hsr.it

Raffaele Ricci is a postgraduate fellow in Broccoli’s lab. He received his Bachelor’s degree in Biological Sciences at Sapienza University of Rome in 2013 and his Master’s degree in Neurobiology at the same University in 2016. In his Bachelor internship he studied alterations of microtubule dynamics in A549cell line due to treatment with silica nanoparticles at CNR in Dr Cundari’s Lab. In his Master internship at EBRI in Prof Cattaneo’s Lab, he worked on Alzheimer’s disease, focusing on the alteration of adult hippocampal neurogenesis due to the presence of intracellular Aβ oligomers in Tg2576 mice. Then he started a post-graduate Erasmus+ project in Spain at CRG in Barcelona in Dr Payer’s Lab. There, he worked on X-chromosome reactivation during miPSCs reprogramming. Now, in Broccoli’s Lab, he is working in setting an innovative gene therapy approach to treat Dravet Syndrome.

Greta Rossi

rossi.greta@hsr.it

Greta Rossi obtained her Bachelor’s degree in Medical and Diagnostic Biotechnology (2015) at the University of Florence and then moved to Milan to attend the Cellular and Molecular Biotechnology Master degree in Vita-Salute San Raffaele University. She’s currently attending her Master internship performing a functional analysis on HeLa cells bearing a recently PD-associated mutation identified in patients with early onset disease, to get new insights into Parkinson’s pathophysiologic mechanisms.

Alessia Salamone

salamone.alessia@hsr.it

Alessia Salamone received her Ph.D. in Pharmacology and Toxicology in 2013 from University of Genova. Her work was focused on the study of the mechanisms of neurotransmitters release modulated by nicotinic receptors. In 2016, she earned a 3-year postdoctoral fellowship to work in Dr. Annamaria Vezzani’s lab (Mario Negri Institute, Milan) and she focused on developing new drug treatments against molecular mechanisms involved in the onset and propagation of seizures in epilepsy. Currently in Dr. Vania Broccoli’s lab, she works on Dravet Syndrome, a severe childhood epileptic disorder, exploring the symptomatic reversibility of Dravet Syndrome in murine models.

Giorgia Tanzi

tanzi.giorgia@hsr.it

Giorgia Tanzi received his Bachelor’s degree in Sciene Biologiche and MD in Molecular Biology of the Cell at Università Statale di Milano. She performed her Master thesis in Prof. Elena Cattaneo’s laboratory of Stem Cell Biology and Pharmacology of Neurodegenerative Diseases. Now, in Dott. Broccoli's Lab, she is working on molecular mechanisms and gene therapy of Dravet Symdrome.

Nicholas Valassina

valassina.nicholas@hsr.it

Nicholas Valassina received his Bachelor’s degree in Medical and Pharmaceutical Biotechnology at Vita-Salute San Raffaele University of Milan in 2017 and his Master’s degree in Medical, Celluar and Molecular Biotechnology at the same University in 2020. In his Master internship in Broccoli’s Lab, he worked on Dravet Syndrome, focusing on the evaluation of its phenotypic reversibility. Currently, he is a postgraduate fellow in Broccoli's lab continuing to investigate on Dravet Syndrome.

Mattia Zaghi

zaghi.mattia@hsr.it

Mattia Zaghi is a master thesis student in Vania Broccoli’s Lab. He received is Bachelor degree in medical Biotechnology from Vita-Salute San Raffaele university in 2014. Currently is working with Alessandro Sessa (thesis supervisor) on assessing the function of Setd5, a histone methyl-transferase, that has been recently found associated with autism and intellectual disability and also is role during normal neural developement.

Alumni

Maxim Bespalov, PhD — 3i Stem Cells, Faculty of Medicine, University of Helsinki, Finland

Tommaso Cabassi — MolMed, Italy

Massimiliano Caiazzo, PhD — Assistant Professor, Universiteit Utrecht, Germany

Cinzia Cancellieri, PhD — Istituto FIRC di oncologia molecolare (IFOM), Italy

Ernesto Ciabatti, PhD — Laboratory of Molecular Biology, Medical Research Council, UK

Maria Teresa Dell’Anno, PhD — Yale School of Medicine, USA

Claudia Di Berardino — Universidad de Valparaíso, Chile

Bruno Di Stefano, PhD — Harvard Department of Stem Cell & Regenerative Biology, USA

Luca Massimino, PhD — Humanitas University, Italy

Marzia Indrigo, PhD — San Raffaele Scientific Institute, Italy

Giuseppina Mastrototaro, PhD — MolMed, Italy

Pietro Giuseppe Mazzara, PhD — Scripps Research Institute, USA

Giuseppe Morabito, PhD — Italy

Alessandro Papale, PhD — San Raffaele Scientific Institute, Italy

Sara Ricciardi, PhD — National Institute of Molecular Genetics, Italy

Alessandro Prigione, PhD — Max Delbrück Center for Molecular Medicine, Germany

Ilda Theka, PhD — Centre for Genomic Regulation, Spain

Federica Ungaro, PhD — Humanitas University, Italy

Support

Vania Broccoli Laboratory gratefully acknowledges grant support from the following institutions:

The laboratory of “Stem Cells and Neurogenesis”, Division of Neuroscience, San Raffaele Scientific Institute is seeking for scientific personnel to expand current projects on Parkinson’s disease and neuronal reprogramming. The lab has established new procedures to convert skin fibroblasts into functional neurons by the expression of a set of reprogramming transcription factors (Caiazzo et al., Nature 476:224-7, 2011; Dell’Anno et al. JCI, 2014 In press). We have conceived a new approach to delay and arrest Parkinson’s pathology and wish to investigate a specific molecular mechanism contributing to disease progression. In addition, we wish to strength our direct neuronal reprogramming approach starting from adult human somatic cells (fibroblasts and blood cells) and explore their therapeutic potential in an animal model of Parkinson’s disease. The laboratory is strategically located within the scientific environment of the San Raffaele Biomedical Area.

Interested candidates should send a detailed curriculum vitae by e-mail to: broccoli.vania@hsr.it