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Laboratory of Molecular and Cellular Pharmacology of Purinergic Transmission  

Lab. Abbracchio

 

Head  Prof.ssa Maria Pia Abbracchio,  mariapia.abbracchio@unimi.it 

Team
Stefania Ceruti, Assistant Professor  stefania.ceruti@unimi.it
Marta Fumagalli, Research Associate  marta.fumagalli@unimi.it
Davide Lecca, Research Associate davide.lecca@unimi.it
Chiara Parravicini , Research Associate  chiara.parravicini@unimi.it
Davide Merli, Research Associate  davide.merli@unimi.it
Dorota Wypych, Research Associate  dorota.wypych@guest.unimi.it
Gianluca Menichetti, Research Associate  gianluca.menichetti@guest.unimi.it
Elisabetta Bonfanti, Ph.D. Student  elisabetta.bonfanti@unimi.it
Marta Boccazzi, Ph.D. Student  marta.boccazzi@unimi.it
Giulia Magni, Ph.D. Student   giulia.magni@guest.unimi.it
Giusy Coppolino, Research fellow  giusy.coppolino@guest.unimi.it
Davide Marangon, Research fellow  davide.marangon@guest.unimi.it

 

Research lines

Our research is aimed at unveiling the pathophysiological roles of extracellular nucleosides and nucleotides and their receptors, the P1 and the P2 receptors. These signalling molecules are widely involved in the function of the nervous, cardiovascular and immune systems, and their specific roles in modulation of cell growth, differentiation, survival and death is increasingly attracting the interest of the scientific community. In this respect, the Laboratory has recently developed an expertise in the purinergic regulation of the activity and function of adult stem-like cells in both the heart, brain and spinal cord, under normal and inflammatory conditions such as those associated to heart infarction, brain and spinal cord ischemia or trauma, and chronic neurodegenerative conditions. Our final aim is to provide the background for setting up novel purinergic based-regenerative approaches to currently incurable human diseases, including heart failure, stroke, Alzheimer´s, and demyelinating diseases. The Laboratory also runs a project aimed at understanding the role of the purinergic system in some forms of neuroinflammatory and neuropathic pain such as migraine.

  1. In vitro and in vivo molecular, pharmacological and biochemical characterization of P1 and P2 receptor, with focus on the cloning, expression and functional characterization of G-protein-coupled P2Y receptors. Some of these aspects are studied in collaboration with the group of prof. Claudia Martini and dr. Letizia Trincavelli of the University of Pisa and with dr. Claudia Verderio e Patrizia Rosa of the CNR Institute of Neuroscience, Milan.
  2. Regulation of cell death processes (apoptosis and necrosis) by purine nucleotides and nucleosides in both brain and in cardiomyocytes, and identification of underlying molecular mechanisms. We are, in particular, interested in defining the role of P2 receptors in brain and heart ischemia (in collaboration with prof. Elena Tremoli, Magnetic Resonance Imaging Unit and Centro Cardiologico Monzino, Milan).
  3. Regulation of the proliferation, differentiation fate, migration, and survival of adult neural stem cells by extracellular nucleotides (in collaboration with dr. Annalisa Buffo, University of Turin).
  4. Role of the P2Y-like receptor GPR17, a new marker of quiescent NG2-positive progenitor cells populating the adult nervous tissue, in reactions to brain injury and demyelinating lesions in rodent models of brain ischemia and multiple sclerosis, and effects of its pharmacological modulation (in collaboration with dr. Roberto Furlan, Ospedale San Raffaele, Milan).
  5. Molecular modeling of G-protein-coupled P2Y receptors and in silico high throughput screening, with special focus on the discovery, design and optimization of new specific pharmacological agents for targeting demyelinating and neuroinflammatory diseases (in collaboration with dr. Ivano Eberini).
  6. Characterization of GPR17 in human traumatic brain injury (in collaboration with dr. Maria Grazia De Simoni, IRCCS-Istituto di Ricerche Farmacologiche “Mario Negri” and dr. Heike Franke, University of Leipzig, Germany).
  7. Genic regulation and effect of microRNAs in GPR17 mediated oligodendroglial maturation and their changes in demyelinating disorders.
  8. Characterization of transgenic reporter GPR17-iCre ERT2 mice (in collaboration with dr. Leda Dimou and prof. Magdalena Götz, University of Munich, Germany).
  9. Role of extracellular nucleotides and P2 receptors in migraine and in trigeminal pain transmission (in collaboration with prof. Andrea Nistri at the International School for Advanced Studies in Trieste and with prof. Peter T. Ohara, UCSF School of Medicine, San Francisco).

 

Techniques

- Cellular and molecular biology (eg. cloning, gene silencing, siRNA, miRNA, mutagenesis, qRT-PCR)

- Cell cultures, isolation and maintenance of primary cultures of astrocytes, oligodendrocyte precursors and microglia and isolation of neurospheres from subventricular zone

- Cytology and histology (eg. in situ hybridization, immunocytochemistry, immunohistochemistry)

- Microscopy techniques (eg. confocal microscopy, time lapse videomicroscopy) and calcium imaging

- Biochemical and functional assays (eg. luciferase activity, in vitro hypoxia/ischemia and cytotoxicity assays)

- Bioinformatics, molecular modeling and computational chemistry

- Murine experimental autoimmune encephalomyelitis (EAE) model

- CFA-Induced allodynia (Freund's Complete Adjuvant) in murine model and von Frey Measurements

- Intra-trigeminal injection of dsRNA in rodents

- Infra-orbital nerve ligation in a rodent model of inflammatory trigeminal pain

 

Selected publications

  1. Ceruti S, Viganò F, Boda E, Ferrario S, Magni G, Boccazzi M, Rosa P, Buffo A, Abbracchio MP (2011) Expression of the new P2Y-like receptor GPR17 during oligodendrocyte precursor cell maturation regulates sensitivity to ATP-induced death. Glia 59:363-78.
  2. Fumagalli M, Daniele S, Lecca D, Lee PR, Parravicini C, Fields RD, Rosa P, Antonucci F, Verderio C, Trincavelli ML, Bramanti P, Martini C, Abbracchio MP (2011) Phenotypic changes, signaling pathway, and functional correlates of GPR17-expressing neural precursor cells during oligodendrocyte differentiation. J Biol Chem286:10593-604.
  3. Ceruti S, Villa G, Fumagalli M, Colombo L, Magni G, Zanardelli M, Fabbretti E, Verderio C, van den Maagdenberg AM, Nistri A, Abbracchio MP (2011) Calcitonin gene-related peptide-mediated enhancement of purinergic neuron/glia communication by the algogenic factor bradykinin in mouse trigeminal ganglia from wild-type and R192Q Cav2.1 Knock-in mice: implications for basic mechanisms of migraine pain. J Neurosci31:3638-49.
  4. Calleri E, Ceruti S, Cristalli G, Martini C, Temporini C, Parravicini C, Volpini R, Daniele S, Caccialanza G, Lecca D, Lambertucci C, Trincavelli ML, Marucci G, Wainer IW, Ranghino G, Fantucci P, Abbracchio MP, Massolini G (2010) Frontal affinity chromatography-mass spectrometry useful for characterization of new ligands for GPR17 receptor. J Med Chem53:3489-501.
  5. Ceruti S, Villa G, Genovese T, Mazzon E, Longhi R, Rosa P, Bramanti P, Cuzzocrea S, Abbracchio MP (2009) The P2Y-like receptor GPR17 as a sensor of damage and a new potential target in spinal cord injury. Brain132:2206-18.
  6. Di Virgilio F, Ceruti S, Bramanti P, Abbracchio MP. Purinergic signalling in inflammation of the central nervous system (2009). Trends Neurosci32:79-87.
  7. Lecca D, Trincavelli ML, Gelosa P, Sironi L, Ciana P, Fumagalli M, Villa G, Verderio C, Grumelli C, Guerrini U, Tremoli E, Rosa P, Cuboni S, Martini C, Buffo A, Cimino M, Abbracchio MP (2008) The recently identified P2Y-Like receptor GPR17 is a sensor of brain damage and a new target for brain repair. PLoS ONE3: E3579.
  8. Ciana P, Fumagalli M, Trincavelli ML, Verderio C, Rosa P, Lecca D, Ferrario S, Parravicini C, Capra V, Gelosa P, Guerrini U, Belcredito S, Cimino M, Sironi L, Tremoli E, Rovati GE, Martini C, Abbracchio MP (2006) The orphan receptor GPR17 identified as a new dual uracil nucleotides/cysteinyl-leukotrienes receptor. EMBO J25:4615-27.
  9. Abbracchio MP, Burnstock G, Boeynaems JM, Barnard EA, Boyer JL, Kennedy C, Knight GE, Fumagalli M, Gachet C, Jacobson KA, Weisman GA (2006) International Union of Pharmacology LVIII: update on the P2Y G protein-coupled nucleotide receptors: from molecular mechanisms and pathophysiology to therapy. Pharmacol Rev5:281-341.
  10. Fumagalli M, Brambilla R, D'Ambrosi N, Volonté C, Matteoli M, Verderio C, Abbracchio MP (2003) Nucleotide-mediated calcium signaling in rat cortical astrocytes: Role of P2X and P2Y receptors. Glia43:218-03.

 

Patents 

1. “Method for the diagnosis of heart disease” United States of America Patent application serial 10/156.352, Filed on May 28, 2002, see: http://da.unimi.it/da/Brevetti/brevetti.doc_cvt.htm#bmk05

2. GPR17 modulators, method of screening and uses thereof” MI2004A 002007 (Oct 21 2004); PCT/EP2005/011157 (Oct 21 2005); U.S.A. 11/665,835 (Nov 11, 2007) granted March 2012

3. “Composti e composizioni che modulano GPR17 e loro usi terapeutici e diagnostici” MI2010A002037 (October 10, 2010), extended internationally on October 10, 2011.

4. GPR17-MODULATING COMPOUNDS, DIAGNOSTIC AND THERAPEUTIC USES THEREOF, PCT/IB20 11/054865

 

 

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