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Cell-surface immobilized vs. internalized magnetic nanoparticles for magnetic hyperthermia studies

Periodic Reporting for period 1 - OUTstandINg (Cell-surface immobilized vs. internalized magnetic nanoparticles for magnetic hyperthermia studies)

Periodo di rendicontazione: 2015-09-01 al 2017-08-31

The MSC OUTstandINg project proposed the use of bioorthogonal click chemistry as tool to covalently attach magnetic nanoparticles (MNPs) to living cell surfaces with the aim to address two fundamental questions in the field of magnetic hyperthermia therapy using MNPs: 1) how the subcellular localization (on the plasma membrane or inside the cells) of MNPs affects their heating behaviour when compared to MNPs in solution 2) how MNPs immobilization and sub-lethal magnetic hyperthermia impact different subcellular signalling pathways and the biophysics of cell membranes.
This multidisciplinary two-years research project was built upon the expertise of the Fellow (Dr. Raluca M. Fratila) in bioorganic chemistry (including click chemistry, bioconjugates and functionalization of surfaces and (nano)materials) and the experience of the supervisor (Dr. Jesús Martínez de la Fuente) and of the host group (GN2 - Nanotherapy and Nanobiosensors, University of Zaragoza, Spain) in nanotechnology and nanomedicine.
The main scientific results achieved can be summarized as follows:
- Successful preparation and characterization of magnetic nanoparticles functionalised with cyclooctynyl derivatives (CO-MNPs) for bioorthogonal click chemistry applications
- Successful covalent immobilization of CO-MNPs on azide surfaces as proof of concept of their bioorthogonal click capability
- Successful installation of azide groups on the membrane of MCF-7 cells
- Successful covalent immobilization of CO-MNPs on the membrane of azide-labelled MCF-7 cells
- Preliminary magnetic hyperthermia studies using CO-MNPs covalently attached on the membrane of azide-labelled MCF-7 cells (experiments carried out during the secondment at University College London, UCL, April-June 2017)
The main training achievements can be summarized as follows:
- New techniques for characterization of MNPs
- New skills in MNP synthesis and functionalization and bioorganic chemistry
- Hands-on training for solution and in vitro magnetic hyperthermia
- New knowledge and skills regarding in vitro experiments (cell culture techniques, cell viability tests, subcellular localization studies of MNPs by confocal fluorescence microscopy)
- Complementary/transferrable skills (proposal writing, Scientific presentation and dissemination skills, project management, leadership, networking)
The action has supposed the launch of a new research line at the host institution in the field of bioorthogonal chemistry applied to nanotechnology. This research line falls into the strategic lines of the Horizon 2020 programme (nanotechnology, advanced materials, health and wellbeing). Future research and collaborations derived from this project are expected to enhance the European competitiveness in the fields of nanotechnology/nanomedicine, bioorthogonal chemistry (most successful research group in the field are located in the USA), and nanomagnetic actuation. New projects based on the preliminary results of OUTstandINg have been already granted to the HI or are under evaluation.
The training-through research carried out in the action had a clear beneficial impact on the career of the researcher, as it allowed Dr Fratila to improve and diversify her competencies and acquire new experimental, managerial, networking and communication skills and to reinforce her professional maturity. Dr Fratila has recently secured a five-year tenure track position at the HI Universidad de Zaragoza through the highly competitive Ramón y Cajal funding scheme from the Spanish Government and has already established a small team formed by one PhD and two MSc students to continue working on the research derived from this MSC project.
Bioorthogonal “click” attachment of MNPs to living cell surfaces