Periodic Reporting for period 1 - GIULIa (MAGNETIC HYPERTHERMIA FOR METASTASIZED TUMOR TREATMENT AND REMOTE MANIPULATION OF MICRODEVICES)
Reporting period: 2023-02-01 to 2025-07-31
In cancer therapy, the heat generated by magnetic nanoparticles (MNPs) is exploited in the so-called magnetic hyperthermia treatment (MHT) to burn primary solid tumors (i.e. in Glioblastoma multiforme patients) and potentiate chemotherapy or radiotherapy. Currently, the accumulation of a sufficient dose of MNPs for MHT at the tumor is only feasible with local intratumorally injection of MNPs. This has limited the MHT therapy only to primary tumors. The MHT treatment of metastasized tumors is still the biggest challenge, until new methods to accumulate the MNP dose for MHT at the metastatic sites are found.
In GIULIa, my ERC consolidator grant, to bring a sufficient dose of magnetic materials to the metastases two distinct routes will be explored: 1. Using biological entities, the natural killer (NK) immune cells equipped with MNPs, to target the metastases; 2. Using an artificial magnetic microdevice with remote locomotion.
Optimal MNPs developed for MHT, will be loaded in/on natural killer (NK) immune cells, and once intravenously injected, they will act as Trojan horses to deliver the right dose of magnetic materials needed for MHT to the metastases. The capability of NK cells and Chimeric Antigen Receptor (CAR) NK immune cells to infiltrate and recognize the tumor will be also studied upon four different infiltrating strategies that will aim at potentiating the immune response of NK immune cells at the tumour. Among these strategies we will identify the best one that merges synergic toxic effects of NK cells immunotherapy with controlled MHT-heat damage of MNPs.
In parallel, we are also addressing the development of magnetic microdevices and their remote locomotion based on MHT-heat gradient, which represent a new technological solution for drug delivery purposes with no tissue-depth attenuation for their actuation. For this part, we are focusing on the development of magnetic based micro-devises which include, as building blocks, metallic magnetic-based heterostructures as hot spots on which to generate bubbles in a liquid and drag an ad hoc designed magnetic microdevices to which, the heterostructures are anchored. There is also a part of the development on the scale-up synthesis of metallic-magnetic nano heterostructures and Janus based microstructures needed as building block for the microdevices.
In GIULIa, my ERC consolidator grant, to bring a sufficient dose of magnetic materials to the metastases two distinct routes will be explored: 1. Using biological entities, the natural killer (NK) immune cells equipped with MNPs, to target the metastases; 2. Using an artificial magnetic microdevice with remote locomotion.
Optimal MNPs developed for MHT, will be loaded in/on natural killer (NK) immune cells, and once intravenously injected, they will act as Trojan horses to deliver the right dose of magnetic materials needed for MHT to the metastases. The capability of NK cells and Chimeric Antigen Receptor (CAR) NK immune cells to infiltrate and recognize the tumor will be also studied upon four different infiltrating strategies that will aim at potentiating the immune response of NK immune cells at the tumour. Among these strategies we will identify the best one that merges synergic toxic effects of NK cells immunotherapy with controlled MHT-heat damage of MNPs.
In parallel, we are also addressing the development of magnetic microdevices and their remote locomotion based on MHT-heat gradient, which represent a new technological solution for drug delivery purposes with no tissue-depth attenuation for their actuation. For this part, we are focusing on the development of magnetic based micro-devises which include, as building blocks, metallic magnetic-based heterostructures as hot spots on which to generate bubbles in a liquid and drag an ad hoc designed magnetic microdevices to which, the heterostructures are anchored. There is also a part of the development on the scale-up synthesis of metallic-magnetic nano heterostructures and Janus based microstructures needed as building block for the microdevices.
During the first half-life of GIULIa project, first, we have established the workplan for ‘Data Organization Strategy’. Upon involvement of the GIULIa teams and my IIT group members, we have estimated for each of the WPs and tasks of GIULIa project the amount of data which will be collected. We have rationally defined the scheme to organize the collection of data, the storage and the preparation of rough data file to be added as supplementary materials for guarantee transparent research for the whole lifetime of GIULIa.
As a second objective, we have aimed at purchasing all the instrumentations accordingly to the assigned GIULIa project fundings. We have established a new Magnetic Particle Imaging (MPI) laboratory at the host institution, the Italian Institute of Technology, in Genoa-Italy, and we have conducted an international tender to purchase the MPI scanner which was finally installed in November 2024 (Figure 1A). This scanner has started to become central for many of ongoing WPs activities of GIULIa ERC project; we have started to perform some measurements on new magnetic materials developed and on magnetic loaded immune cells. We have also acquired a new magnetic hyperthermia device which will enable us to apply the alternated magnetic field, through a movable coil connected to a long arm, from the core of the device (Figure 1B) and which will be used for some magneto-optical characterization of our materials. We also bought a microfluidic device for continuous synthesis of building blocks (Figure 1C) and a microfluidic pump system (Figure 1D), which is capable to simulate the blood vessel/capillary flow on our magnetic microdevices or MNPs-loaded immune cells dispersed in the flow to study their behaviour (Figure 1C) under an optical/fluorescent microscope coupled with the magnetic hyperthermia devise of Figure 1B. Finally, we have also bought a sample positioner as an accessory for our Alternating Current (AC) magnetometer device (Figure 1E). This tool enables to automatically perform the AC magnetometer measurements of magnetic materials exposed to different environments or to perform long-term studies with multiple repetition of measurements of the same sample over prolonged time.
For the scientific specific task of the project, we have set a new protocol to engineering the natural killer (NK) cells with magnetic nanoparticles (MNPs), to be used as trojan horses to deliver, at the tumor. This protocol was studied on NK92 cells and NK cells extracted from healthy donor blood but also a T-cell line and is under investigation on CAR-NK cells. The whole collection of data has been used to file an Italian patent application (Application PT230732 of the 28/03/2024). In parallel, magnetic-heterostructure and plasmonic magnetic heterostructures have been developed as main building blocks for the development of magnetic microdevices to act as microrobot within this first period of GIULIa project. We have been focusing on synthesizing bimetallic dimers and tri-metallic dumbbell heterostructures featuring a magnetic core and gold-silver plasmonic domains and these synthesis conditions, set on bench protocol, are now under further development to obtain the same heterostructures under microfluidic conditions. We have developed a well-optimized, microwave-assisted Fe3O4 NCs synthesis process, which allowed us to produce highly monodisperse, well-crystalline structures in very short synthesis durations which is of the order of minutes (Wid Mekseriwattana et al., https://doi.org/10.1002/adfm.202413514 ). These conditions are now under further implementation for an in-flow synthesis method for the continuous production of ferrite based nanocubes to be then used for the assembly of magnetic micro-device.
As a second objective, we have aimed at purchasing all the instrumentations accordingly to the assigned GIULIa project fundings. We have established a new Magnetic Particle Imaging (MPI) laboratory at the host institution, the Italian Institute of Technology, in Genoa-Italy, and we have conducted an international tender to purchase the MPI scanner which was finally installed in November 2024 (Figure 1A). This scanner has started to become central for many of ongoing WPs activities of GIULIa ERC project; we have started to perform some measurements on new magnetic materials developed and on magnetic loaded immune cells. We have also acquired a new magnetic hyperthermia device which will enable us to apply the alternated magnetic field, through a movable coil connected to a long arm, from the core of the device (Figure 1B) and which will be used for some magneto-optical characterization of our materials. We also bought a microfluidic device for continuous synthesis of building blocks (Figure 1C) and a microfluidic pump system (Figure 1D), which is capable to simulate the blood vessel/capillary flow on our magnetic microdevices or MNPs-loaded immune cells dispersed in the flow to study their behaviour (Figure 1C) under an optical/fluorescent microscope coupled with the magnetic hyperthermia devise of Figure 1B. Finally, we have also bought a sample positioner as an accessory for our Alternating Current (AC) magnetometer device (Figure 1E). This tool enables to automatically perform the AC magnetometer measurements of magnetic materials exposed to different environments or to perform long-term studies with multiple repetition of measurements of the same sample over prolonged time.
For the scientific specific task of the project, we have set a new protocol to engineering the natural killer (NK) cells with magnetic nanoparticles (MNPs), to be used as trojan horses to deliver, at the tumor. This protocol was studied on NK92 cells and NK cells extracted from healthy donor blood but also a T-cell line and is under investigation on CAR-NK cells. The whole collection of data has been used to file an Italian patent application (Application PT230732 of the 28/03/2024). In parallel, magnetic-heterostructure and plasmonic magnetic heterostructures have been developed as main building blocks for the development of magnetic microdevices to act as microrobot within this first period of GIULIa project. We have been focusing on synthesizing bimetallic dimers and tri-metallic dumbbell heterostructures featuring a magnetic core and gold-silver plasmonic domains and these synthesis conditions, set on bench protocol, are now under further development to obtain the same heterostructures under microfluidic conditions. We have developed a well-optimized, microwave-assisted Fe3O4 NCs synthesis process, which allowed us to produce highly monodisperse, well-crystalline structures in very short synthesis durations which is of the order of minutes (Wid Mekseriwattana et al., https://doi.org/10.1002/adfm.202413514 ). These conditions are now under further implementation for an in-flow synthesis method for the continuous production of ferrite based nanocubes to be then used for the assembly of magnetic micro-device.
To translate these basic findings into the two main final aims of GIULIa’s research project, several steps need to be taken. For the immune cell’s activities, we will move towards the in vitro studies to test MNPs loaded NK cell killing response towards tumour cells upon different infiltrating and targeting strategies to optimize their therapeutic outcomes. Simultaneously, we will investigate the magnetic particle imaging (MPI) properties of MNP-labelled immune cells, aiming to generate robust in vitro MPI datasets. These data will support the preparation of regulatory documentation needed to obtain clearance for in vivo preclinical studies, which will be selectively conducted to validate the most promising approaches will uncover the best infiltrating NK cells strategy with the best magnetic nanoparticles formulations. In parallel, besides producing MNPs and magneto-plasmonic heterostructures with precise control over the size, size distribution, shape and composition, their scalable production must be achieved even by exploring fluidic methods. As a next step, we aim to integrate these nanomaterials into Janus-like microdevices, using inorganic or polymeric matrices to achieve structured assemblies. This will pave the way for proof-of-concept studies focused on assessing their locomotion and functional performance.