Prof. Antonino Cattaneo, Director of Bio@SNS, has been granted by the European Human Brain Project, with a project “The systematic generation of antibodies against Neuroligins and their interaction partners: a proof of concept study”. The Neuroantibodies team also includes the European Brain Research Institute (EBRI, Rome) and the Institute of Biophysics CNR (Palermo)
The Human Brain Project is a ten-year project funded by the European Commission as a FLAGship project under Horizon 2020. The project, which will have a total budget of over Euro 1 billion, is European-led with a strong element of international cooperation. The goal of the project is to build a completely new ICT infrastructure for neuroscience, and for brain-related research in medicine and computing, catalyzing a global collaborative effort to understand the human brain and its diseases and ultimately to emulate its computational capabilities.
Antibodies are essential reagents for studying the brain proteome, and antibody assays are an essential toolkit for descriptive and functional analyses, that include localization, isolation, interaction, quantification and interference. The group of A. Cattaneo has pioneered the technology of intracellular antibodies (intrabodies) and the construction of libraries of antibody domains in yeast cells, for the selection of antibodies to be used for subcellularly precise protein silencing (the Intracellular Antibody Capture Technology, IACT). This allows the effective isolation of antibodies directly from gene sequences, with no manipulation of the corresponding proteins.
The objective of the NEUROANTIBODIES Project is to provide a proof-of-concept for the systematic generation of antibodies against a family of synaptic proteins and their proximal interactome, exploiting IACT. This should allow in principle, to streamline the procedure to the high-throughput necessary to address the large numbers of targets, relevant for the HBP. After biochemical and histochemical validation and characterization, the antibodies will be expressed in mouse neurons for functional interference studies, acting at different points of the protein network involved. The experimental readout will involve the electrophysiological evaluation, by patch-clamp recording from CA1 principal cells, of excitatory glutamatergic and inhibitory GABAergic synaptic transmission and long-term plasticity processes, in response to the selective protein interference by the different intrabodies. Unsupervised, automatic computational modeling of the bidirectional excitatory and inhibitory synaptic transmission, involving the receptors and protein network under study, in basal or protein interference conditions, will be implemented and integrated into the HBP Brain Simulation Platform. The planned work will provide a feasibility demonstration of the use of IACT and intrabodies for the functional, quantitative dissection of a synaptically relevant receptor linked protein network.
Project references: FET Flagship CP/CSA, Grant agreement number 604102