The research project proposed in this action aimed to develop targeted and BBB-permeable nanoparticles aiming to provide a new treatment for glioblastoma multiforme. To guide my way toward this aim I set four specific objectives: 1) generation of site-specifically modified antibodies and peptides, 2) preparation and characterization of the targeted polyplexes, 3) in vitro assessment of safety, transfection efficiency, selectivity and BBB transport, and 4) in vivo study of targeting capacity and therapeutic efficacy. In the 24 months given for this action, I have completed goals 1-3. Given the COVID-19 pandemic many of the experiments were delayed and, as a result, objective 4, which involves in vivo experiments has not been accomplished yet. However, the remaining experiments will be conducted within the research projects awarded.
PROGRESS BY OBJECTIVES
Objective 1: Generation of site-specifically modified antibodies and peptides
Antibodies capable of targeting EGFR that can be anchored on the nanoparticles were produced. The extensive research conducted toward this end, led to the production of a review article published in the high impact journal (IF: 14.5) ACS Central Science. As for peptides intended to cross the BBB (BBB-shuttle peptides), not only we synthesized sequences reported in the literature but also we developed novel cyclic protease-resistant anti-transferrin receptor peptides. A research article reporting the novel BBB shuttles developed and a review are in preparation and will be submitted in the last trimester of 2021.
Objective 2: Development of the targeted polyplexes
Acrylate-terminated pBAEs were produced. As mentioned in the project, several strategies were explored in order to coat the pBAEs. Overall, polyplexes with adequate physicochemical properties have been developed.
Objective 3: In vitro assessment of safety, transfection efficiency and BBB transport
pBAEs polyplexes were proved to have negligible toxicity on a variety of cells. After a long process of optimization, the new coating enabled reducing the transfection efficiency of pBAEs by over 24-fold. Upon ligand modification, selective transfection to cancer cells overexpressing the transferrin receptor was enhanced by over 42-fold. Therefore, nanoparticles with selective transfection capacity have been prepared. These results are the basis for a manuscript in preparation. Moreover, we have assayed the capacity of coated nanoparticles in a BBB cell-based model and identified the most suitable candidates. The preliminary results obtained for the transport capacity of the final conjugates is currently being verified.
Objective 4: In vivo study of targeting capacity and therapeutic efficacy
Due to the COVID-19 pandemic all experiments were delayed and we intend to start the in vivo stage in late 2021. We have obtained funding for three additional projects well aligned with this proposal that will enable pursuing these experiments.
DISSEMINATION AND EXPLOITATION
Regarding scientific audiences, so far, we have published a review article at ACS Central Science (IF: 14.5) and we are currently preparing three more manuscripts. Moreover, results from this Project have been/will be shared at four international conferences and a seminars.
As planned activities for the general public, I have given a conference to IQS, delivered a talk followed by a colloquium with high school teachers in the Biosciences talks organized by IQS, and given another talk to pre-university students invited by the Catalan Chemical Society. Additionally, a webpage devoted to the research program that has emerged from this project has been created and an article for the IQS community and affiliated institutions has been prepared and will be released in September.
Regarding exploitation of the results, we have been contacting several companies who could be interested either in the technologies we have developed in the project or in the methods we have set up in the process. We have applied for a competitive project with one of the companies and we are negotiating a contract for the development of peptides against a particular target with another company.
Last but not less important, building on the preliminary results of this project and the experience acquired with this action, I have obtained the three competitive projects. In these projects, we will develop more sophisticated and efficient systems to target the nanomedicine developed in the MSCA-IF fellowship, which should enable higher degree of selectivity.
