Experimental Imaging Center
Drug Screening Technologies
Recent years have witnessed an evolution in the approach to drug discovery. The conventional molecular target-based screening is now combined with cell- based phenotypic approaches, where targets do not need to be known and active lead compounds are selected based on their capability to ameliorate the disease phenotype. In most pathological contexts the intrinsic complexity of the phenotypes, require the use of ad hoc cellular models as well as sophisticated testing technologies.
By taking advantage from long-established competences in, neurophysiology, pharmacology and microscopy, we develop leading-edge solutions for small and medium-scale drug screening programs. In our cell based assays, we develop and employ either immortalized cell lines or cells differentiated from induced pluripotent stem cells (iPSCs) obtained from patients or normal human subjects. This ambitious research program is possible thanks to the long-standing collaboration with research groups from IRCCS Ospedale San Raffaele, Politecnico di Milano as well as biotechnological and pharmaceutical chemical companies.
We have recently patented a new methodology, and developed a prototype, for ion channel automated drug screening programs in intact cells as well as an innovative fluorescence-based approach for membrane potential measurement for functional optical studies. These approaches can be complemented by a drug profiling analysis performed by extracellular electrophysiology: a bioreactor for multielectrode array (MEA) measurements that permits the study of neuronal activity and connectivity under control environmental conditions on multiple samples simultaneously. Our current field of interest is the screening of molecules active on pH sensitive channels (ASIC channels). A drug discovery program, based on rationally designed compounds, is active for the identification of new ASIC channels modulators.
Bencheva LI, De Matteo M, Ferrante L, Ferrara M, Prandi A, Randazzo P, Ronzoni S, Sinisi R, Seneci P, Summa V, Gallo M, Veneziano M, Cellucci A, Mazzocchi N, Menegon A, Di Fabio R. Identification of Isoform 2 Acid-Sensing Ion Channel Inhibitors as Tool Compounds for Target Validation Studies in CNS. ACS Med Chem Lett. 2019 Feb 7;10(4):627-632. doi: 10.1021/acsmedchemlett.8b00591. eCollection 2019 Apr 11.PMID: 30996808 Free PMC article.
Spinelli A, Girelli M, Arosio D, Polito L, Podini P, Martino G, Seneci P, Muzio L, Menegon A. Intracisternal delivery of PEG-coated gold nanoparticles results in high brain penetrance and long-lasting stability. J Nanobiotechnology. 2019 Apr 3;17(1):49. doi: 10.1186/s12951-019-0481-3.PMID: 30943991 Free PMC article.
Mazzocchi N, Grohovaz F, Taverna S, Menegon A. Membrane potential changes occurring upon acidification influence the binding of small-molecule inhibitors to ASIC1a. Neuropharmacology. 2019 Apr;148:366-376. doi: 10.1016/j.neuropharm.2019.01.033. Epub 2019 Feb 1.PMID: 30716415
Menegon A, Pitassi S, Mazzocchi N, Redaelli L, Rizzetto R, Rolland JF, Poli C, Imberti M, Lanati A, Grohovaz F. A new electro-optical approach for conductance measurement: an assay for the study of drugs acting on ligand-gated ion channels. Sci Rep. 2017 Mar 21;7:44843
Mazzocchi N, De Ceglia R, Mazza D, Forti L, Muzio L, Menegon A. Fluorescence-Based Automated Screening Assay for the Study of the pH- Sensitive Channel ASIC1a. J Biomol Screen. 2016 Apr;21(4):372-80
de Ceglia R, Chaabane L, Biffi E, Bergamaschi A, Ferrigno G, Amadio S, Del Carro U, Mazzocchi N, Comi G, Bianchi V, Taverna S, Forti L, D'Adamo P, Martino G, Menegon A, Muzio L. Down-sizing of neuronal network activity and density of presynaptic terminals by pathological acidosis are efficiently prevented by Diminazene Aceturate. Brain Behav Immun. 2015 Mar;45:263-76
Biffi E, Regalia G, Ghezzi D, De Ceglia R, Menegon A, Ferrigno G, Fiore GB, Pedrocchi A. A novel environmental chamber for neuronal network multisite recordings. Biotechnol Bioeng. 2012 Oct;109(10):2553-66
Caiazzo M, Dell'anno MT, Dvoretskova E, Lazarevic D, Taverna S, Leo D, Sotnikova TD, Menegon A, Roncaglia P, Colciago G, Russo G, Carninci P, Pezzoli G, Gainetdinov RR, Gustincich S, Dityatev A, Broccoli V. Direct generation of functional dopaminergic neurons from mouse and human fibroblasts. Nature. 2011 Jul 3;476(7359):224-7
Rossi S, Muzio L, De Chiara V, Grasselli G, Musella A, Musumeci G, Mandolesi G, De Ceglia R, Maida S, Biffi E, Pedrocchi A, Menegon A, Bernardi G, Furlan R, Martino G, Centonze D. Impaired striatal GABA transmission in experimental autoimmune encephalomyelitis. Brain, Behavior, and Immunity. 2011; Jul;25(5):947-56
Menegon A, Bonanomi D, Albertinazzi C, Lotti F, Ferrari G, Kao H.T, Benfenati F, Valtorta F. Protein kinase A-mediated synapsin I phosphorylation is a central modulator of Ca2+-dependent synaptic activity. J Neuroscience. 2006; Vol. 26 (45):11670-81
Bonanomi D, Menegon A, Miccio A, Ferrari G, Corradi A, Kao H, Benfenati F, Valtorta F. Phosphorylation of synapsin I by cAMP-dependent protein kinase controls synaptic vesicle dynamics in developing neurons. J. Neuroscience. 2005; Vol. 25(32) pp 7299-308
Menegon A, Verderio C, Leoni C, Benfenati F, Czernik A.J, Greengard P, Matteoli M, Valtorta F. Spatial and temporal regulation of Ca2+/calmodulin dependent protein kinase II activity in developing neurons. J. Neuroscience. 2002; Vol. 22, pp 7016-26
Menegon A, Dunlap D, Castano F, Benfenati F, Czernik A.J, Greengard P, Valtorta F. Use of phosphosynapsin I-specific antibodies for image analysis of signal transduction in single nerve terminals. J. Cell Sci. 2000; Vol 113, pp 3573-82