IRFMN - Mario Negri Institute for Pharmacological Research IRCCS, Milano, Italy

Lead beneficiary of WP6 - Capture-Enhanced Neutron Irradiation (CENI) Effectiveness.
IRFMN is the main partner of NECTAR consortium with the experience and know-how to handle properly and effectively Aβ proteins as well as AD animal models and related healthy counterparts. For this reason, IRFMN is first of all the partner in charge of Aβ procurement, synthesis and preparation for all the experiments, from the level of water solutions up to animals.
In addition, IRFMN is responsible for: (i) APP/PS1 mouse purchasing and maintenance, (ii) APP/PS1 mouse evaluations at behavioral and neuropathological level upon neutron irradiation, (iii) evaluations of residual neurotoxicity of irradiated Aβ aggregates on cell cultures.
To go specifically through the tasks of the project, IRFMN will be the leader partner for the following tasks: T3.1 (Aβ aggregates characterization), T4.2 (Aβ depolymerization by CENI), T4.3 (residual toxicity of Aβ aggregates in vitro and in vivo), T5.2 (in vivo CENI toxicity), T6.1 (in vivo CENI effectiveness); while IRFMN will be a participant in the following steps: T2.4 (Aβ binding assay through SPR).
Due to their general objectives, IRFMN is involved also in the management (T1.1-T1.3) of the project as well as the dissemination, communication and training activities (T7.1-T7.4).

Experience in: IRFMN has a long lasting experience with in-vivo studies, having analyzed and characterized several animal models of AD both at behavioral and neuropathological level. Importantly, several therapeutic approaches were previously tested on AD mice, which implies a large experience with the assessment of therapeutic efficacy at multiple level within the complex, multi-factorial pathological scenario of AD. IRFMN has also a very large experience with peptide synthesis including Aβ, the characterization and analysis of Aβ aggregate formation and the determination of changes within the peptide aggregation process. Furthermore, IRFMN will share its expertise with SPR technique, allowing the assessment of protein binding and affinity.

Facilities and tools: The Mario Negri Institute is fully equipped with all the facilities, know-how and technologies required to execute the experiments described in the project. IRFMN disposes of: stereotaxic and gas anesthesia systems for brain surgery; various behavioral apparatus connected to the Noldus system; three locations with cryostats for brain slice collection. Two Olympus confocal microscopes and one Nikon structured illumination microscope for immunofluorescence imaging. Two light microscopes supported by the analysis program for histological analysis. One virtual stage Nikon microscope for image acquisition. Two work stations with the IMARIS software for imaging post-processing. Imaging systems for quantitative analysis of Western blots. The Institute provides also a very large high-standard Specific Pathogen Free animal facility. For in vitro experiments the department is fully equipped for chemical and solvent storage and handling, cell culture maintenance, several cell-culture laminar flow hoods and incubators. It also includes an advanced equipment for cell sorting (FACS), biochemical analysis (absorption and fluorescence plate readers) and microscopy (optical e confocal microscopes, epifluorescent microscope equipped with thermostatic chambers for time-lapse live imaging).

IRFMN - Mario Negri Institute for Pharmacological Research IRCCS, Milano, Italy

Letizia Dacomo

Marco Gobbi

Edoardo Micotti