Co-culture of rat brain cells as a tool for studying cell-cell interactions Gabriella Schiera *, Patrizia Proia**, Carlo Maria Di Liegro*, Giovanni Savettieri***, Marianna Lo Pizzo*, Patrizia Saladino*, and Italia Di Liegro*** Departments of *Scienze e Tecnologie Molecolari e Biomolecolari, **Studi Giuridici, Economici, Biomedici, Psicosociopedagogici delle Scienze Motorie e Sportive, and ***Biomedicina Sperimentale e Neuroscienze Cliniche, University of Palermo, Palermo, Italy Brain capillary endothelial cells (BCECs) form the blood-brain barrier (BBB) in response to interaction with other brain cells (astrocytes, pericytes and neurons). BCECs are characterized by tight junctions (TJ), maturation and stabilization of which require different proteins, such as occludin. When co-cultured with astrocytes and neurons, BCECs were found to form a monolayer resembling the natural BBB: paracellular flux of dopamine and sucrose (i.e. compounds which are unable to cross the BBB in vivo) significantly decreased (1), while the transendothelial electrical resistance (TEER) increased. In these conditions, BCECs produced a larger amount of occludin and tended to localize it at the cell periphery, thus suggesting formation of TJs (1). Since we also discovered that oligodendroglioma cells shed extracellular membrane vesicles (MVs; 2), we investigated whether also neurons and/or astrocytes can release MVs and whether these vesicles contained angiogenic factors. The results of these analyses demonstrated that all kinds of brain cells actually shed MVs containing FGF-2 and VEGF (3-4). On the basis of these findings, we investigated the possibility that the BBB model could be used to study the molecular events that result in BBB damage, in some pathological conditions, such as, for example, multiple sclerosis (5). We are now investigating whether cultured astrocytes shed vesicles containing aquaporin 4 (AQP4), a protein which has been involved in brain edema. Our results suggest that production of AQP4 increases in stressed astrocytes. 1. Schiera G. et al., 2005, J Cell Mol Med 9: 373-9 2. D’Agostino S. et al., 2006, Int J Oncol 29: 1075-85 3. Schiera G et al. 2007, J Cell Mol.Med 11: 1384-94 4. Proia P. et al. 2008, Int J Mol Med 21: 63-7 5. Proia et al. 2009, Int J Mol Med 24: 743-7 --------- Pharmacophore modelling as useful tool in the lead compounds identification and optimization Marco Tutone , Licia Pantano, Antonino Lauria, Annamaria Martorana, Anna Maria Almerico Dipartimento di Scienze e Tecnologie Molecolari e Biomolecolari (STEMBIO) Sezione di Chimica Farmaceutica e Biologica The goal of computer-aided molecular design methods in modern medicinal chemistry is to reduce the overall cost and time associated to the discovery and development of a new drug by identifying the most promising candidates to focus the experimental efforts on. Very often, many drug discovery projects have reached already a well-advanced stage before detailed structural data on the protein target have become available. A possible consequence is that often, medicinal chemists develop novel compounds for a target using preliminary structure–activity information, together with the theoretical models of interactions. Only responses that are consistent with the working hypothesis contribute to an evolution of the used models. Within this framework, the pharmacophore approach has proven to be successful, allowing the perception and understanding of key interactions between a receptor and a ligand[1]. In recent years, our research group exploited this useful modeling tool with the aim to identify new chemical entities and/or optimizing known lead compounds to obtain more active drugs in the field of antitumor, antiviral, and antibacterial drugs. In this communication, we present an overview of our recent works in which we used the pharmacophore modelling approach combined with induced fit docking, 3D-QSAR approach, and HTVS for the analysis of drug-receptor interactions and the discovery of new inhibitors of IKKβ, Bcl-xl, and c-kit tyrosine kinase, all targets involved into the initiation and the development of different types of cancer[2-5]. brought to you by CORE View metadata, citation and similar papers at core.ac.uk provided by Archivio istituzionale della ricerca - Università di Palermo