The Partners of the NEVERMIND Consortium have worked for the first 18 months of the project with their complementary expertise in different scientific fields, starting from the synthesis of nanovectors (NVs) to their physico-chemical and biological characterization.
The collaboration between the Univeristy of Milan (UNIMI) and the University of Milano-Bicocca (UNIMIB) led to the SYNTHESIS of different drug loaded-NV formulations functionalized with modified peptides that allow the crossing of the blood brain barrier and the release of the drugs in the neuroinflammatory or tumor niche.
⇒ UNIMI designed and synthetized two protease target peptides that have been inserted in NV structures
⇒ UNIMIB has prepared 3 stable NVs preparations that were characterized for their chemical and physical properties, and the evaluation of NVs capacity to be internalized in the cells has been tested.
The Laboratory of Nanomedicine and Clinical Biophotonics (LABION) at Fondazione Don Carlo Gnocchi has worked on the validation of drug-loaded NVs.
⇒ LABION performed the QUALITY CONTROL of NVs, studying at the same time the encapsulated candidate drug release and the target specificity through the correct surface functionalization, taking advantages of the biophotonics-based tecniques Raman Spectroscopy and Surface Plasmon Resonance imaging (SPRi).The Raman analysis allowed to verify the encapsulation of the drugs and the presence of modified peptides and mApoE on the NV surface in a label-free manner that prevents the introduction of artifacts prior to their testing in the in vitro models.
⇒ LABION has also verified the INTERACTIONS of mApoE with target receptors for the blood brain barrier crossing and the integrity of the encapsulated drugs, providing information regarding the release dynamics, that could help the design of the next in vitro and in vivo studies, clarifying the binding kinetic and the candidate drug release profile.
Fondazione Don Carlo Gnocchi has also studied the IMMUNOLOGY and EPIGENETIC aspects for the evaluation of the biological effects of drug-loaded NVs in Alzheimer’s patients.
⇒ The Laboratory of Molecular and Translational Medicine have investigated the response of immunce cells to the candidate NVs. The results can contribute to the definition of the dose necessary to obtain a significant effect on the specific molecular pathway. Their experience in genetic profiling has also allowed to detect NVs induced variations in the expression of specific gene and miRNA related to neuroinflammation.
Ospedale San Raffaele (OSR) and Istituto Clinico Humanitas (ICH) have started the series of experiments aimed at testing the BIOCOMPATIBILITY and TOXICITY of the NVs.
⇒ OSR has investigated the biocompatibility of the formulations that have been synthetized from the other partners comparing the effects of NVs and free drugs in neurons finding that they are not toxic for cells.
⇒ ICH has set up the experimental setting to test the toxicity of drug-loaded NVs on glioblastoma cells obtained from tumors excised from patients.