WP1 Coordinators
The coordinators for the WP1 are: G. I. Forte and E. Scifoni
WP1 Participants
The WP1 Participants are as listed in the following table:
| Name | Unit | FTE | Name | Unit | FTE | Name | Unit | FTE |
| CNR Grant Holder | TIFPA | 0.4 | Cella Zanacchi F. | PI | 0.1 | Minafra L. | LNS | 0.1 |
| Attili A. | TIFPA | 0.2 | Cordoni F. | TIFPA | 0.3 | Monti V. | TO | 0.3 |
| Bellinzona V.E. | TIFPA | 0.2 | Costa M. | PI | 0.1 | Russo G. | LNS | 0.5 |
| Bisio A. | TIFPA | 0.5 | Croci S. | TIFPA | 0.6 | Scifoni E. | TIFPA | 0.3 |
| Bortolussi S. | MI | 0.2 | Ficarra M. | LNS | 1 | Sokol O. | TIFPA | 0.1 |
| Boscolo D. | TIFPA | 0.1 | Forte G.I. | LNS | 0.7 | Strettoi E. | PI | 0.1 |
| Bravatà V. | LNS | 0.5 | Fuss M. | TIFPA | 0.1 | Tinganelli W. | TIFPA | 0.1 |
| Calvaruso M. | LNS | 0.5 | La Tessa C. | TIFPA | 0.2 | Tommasini F. | TIFPA | 0.2 |
| Cammarata F. | LNS | 0.1 | Manghi M. | TIFPA | 0.5 | Vannini E. | PI | 0.1 |
Main Goals
The WP1 activity is devoted to gaining insights in understanding the still poorly explained FLASH effect mechanism. For this goal, the WP1 is articulated in 4 main tasks:
Task 1 - Biophysical modeling
Different approaches will be used to analyze the spatiotemporal damage evolution at high dose rates, to obtain a dose enhancement factor, dependent on all the relevant parameters connected to the FLASH effect, to realize a biologically driven TPS. The modeling will explore:
i) the heterogeneous chemical stage, by expanding the TRAX-CHEM Monte Carlo chemical track structure code;
ii) the homogeneous chemical stage, by plugging an analytical model of tissue O2 diffusion-reaction;
iii) The DNA repair kinetics in Flash/Conv conditions;
iv) advanced Normal Tissue Complication Probability modelling as a function of different irradiation and target condition.
Task 2 - In vitro Experiments
The in vitro experiments aim to highlight biological processes activated by flash mode irradiation, to perform a “tuning” of physical irradiation parameters (dose, dose/rate, dose/pulse), selecting the combinations that enable to appreciate the effect at microscopic level. Two breast cell lines, the non-tumorigenic MCF10A and the tumorigenic MDA-MB-231, will be used and experiments will be performed varying doses, dose rates and oxygen concentration, to evaluate the following biological endpoints: survival and DNA damage; cell death (apoptosis/necrosis) and senescence; cells redox status; profiling of secreted immunological markers; RNA-seq on irradiated cells; microtubule integrity and collagen structure modification studies. The feasibility of using the organ-on-a-chip technology will be investigated, to provide insights into normal human organ functions. This activity will be carried out by CT (in-kind) in collaboration with the University of Surrey.
Task 3 - Ex vivo Experiments
Starting from retinal cell cultures ARPE19 cells (a model of retina-pigment epithelium) the effect of FLASH/CONV approaches identifying effective and damaging doses will be studied. Then, whole mouse retinas will be exposed to FLASH/CONV irradiation. Endpoints evaluated will be: local microglia activation, apoptosis and oxidative stress.
Task 4 - Upgrade of UNITO Linac and TIFPA p-Lab
TO unit will modify a conventional clinical ELEKTA LINAC (4-18 MeV) of TO and Physics Department (UNITO), fully dedicated to research. The modification will allow delivering electron beams achieving FLASH RT rates, controlling electron pulses, beam output stability, pulse constancy and beam flatness. The dosimetric calibration will be performed in collaboration with the CT unit. The upgraded LINAC will be used to test beam monitors/dosimeters. The TIFPA team will upgrade, in collaboration with IBA, the proton beam experimental facility to deliver ultra-high dose rates.
WP1 Deliverables
The WP1 Deliverables are summarized in the following table:
| Deliv. | Short Name | Description | When (M) | Deliv. | Short Name | Description | When (M) |
| D1.1 | RadChem Modelling | Report of radchem extension to the heterogeneous stage. | 1-16 | D2.4 | Radbio consolidation and insights | Repetition of some radiobiological experimental sets requiring statistical robustness or potential irradiation for selected end points using other beams with different characteristics. | 24-36 |
| D1.2 | DDRK Modelling | Report on DDRK Modelling studies. | 1-24 | D2.5 | Radbio on 3D under FLASH and CONV | Report on the effects of CONV vs. FLASH irradiation on the 3D collagen scaffold. Data collected mainly with 2nd harmonic/two photon microscopy. | 1-24 |
| D1.3 | DMF Modelling | Report on DMF Modelling studies. | 16-36 | D2.6 | Radbio on cytoskeleton under FLASH and CONV | Report on the effects of CONV vs. FLASH irradiation on the cytoskeleton (i.e microtubules) fiber integrity. Data collected mainly with advanced microscopy techniques. | 1-32 |
| D2.1 | Radbio under Conventional Irradiation | Report on Cell survival/DNA repair and redox status under conventional irradiation. Collecting of conventionally irradiated medium and pellets for future immunological profiling and RNAseq. | 1-12 | D3.1 | Retina Cells | Report on tumor cells and retina-pigment cells studies. | 16 |
| D2.2 | Radbio under Flash Irradiation | Report on Cell survival/DNA repair and redox status under Flash irradiation. Collecting of conventionally irradiated medium and pellets for future immunological profiling and RNAseq. | 13-20 | D3.2 | Retina explanted | Report on explanted mouse retina. | 24 |
| D2.3 | Radbio profiling | Immunological profiling and RNAseq. | 13-24 | D4.1 | TO-Linac dosimetric calibration | The dosimetric calibration of the INFN Torino LINAC, after its modification, will be performed in collaboration with Catania section. | 24 |
| D4.2 | TIFPA-pFLASH dosimetric calibration | The dosimetric calibration of the TIFPA FLASH beam, after its modification, will be performed in collaboration with PI and CT. | 24 |