Description
Pavol Lukáč1,2,, Pavel Bláha3, Leonid Sushytskyi1, Lenka Rajsiglová1, Paolo Tenti1,2, Dmitry Stakheev1,4, Daniel Smrž1,4, Lorenzo Giuffrida3, Daniele Margarone3, Luca Vannucci1,
- Presenting Authors
1 Laboratory of Immunotherapy, Institute of Microbiology of the CAS v.v.i., Prague, Czech Republic
2 Department of Cells Biology, Faculty of Science, Charles University, Prague, Czech Republic
3 ELI Beamlines Facility, The Extreme Light Infrastructure ERIC, Dolní Břežany, Czech Republic
4 Department of Immunology, Second Faculty of Medicine, Charles University and University Hospital Motol, Prague, Czech Republic
Increasing knowledge on cancer biology and immunology together with an exponential growth in biotechnologies and new technologies in physics and chemistry are revolutionizing cancer diagnosis and treatments. New technologies are helping us to find more effective solutions for new therapeutic procedures in the treatment of the increasingly frequent occurrence of patients with cancer. Radiation therapy (accelerated photons, neutrons, electrons, protons, and ions) is in growing interest not only for its possibilities to directly affect the tumor cells, but also to promote valuable and specific immune response. To find new approaches in radiation treatments, the ELI Beamlines facility (Dolní Břežany, CZ) develops novel technologies which explore application of advanced laser-driven accelerators for anticancer treatments as an alternative to traditional accelerator types. The laser-driven proton beams are produced with specific temporal and dose-rate characteristics (very high peak dose-rate beams pulsed in nanosecond delivery times) that may potentially offer novel and increased treatment benefits including better sparing of healthy tissue, and cancer stress molecule and cell debris release with elicitation of active immune responses. The laser acceleration system is powered by the L3 HAPLS petawatt laser, integrated with the ELIMAIA (ELI Multidisciplinary Applications of laser-Ion Acceleration) laser-plasma accelerator and the ELIMED (ELI MEDical application) beam transport and dosimetry line, enabling multi-shot proton irradiation. In our experiments, we irradiated spheroids of human colorectal adenocarcinoma (three-dimensional cell culture model). Spheroids, with their more complex structure than cell monolayers, are more suitable for studying the tumor microenvironment responses. In the study, we used 6-day-old spheroids, monitoring changes in 24 hours after irradiation using the Live-Dead Assay on confocal microscopy. The results showed cellular damage to the nuclei after irradiation and the beginning of spheroid disintegration. Immunogenic cell death and stress molecules expression are under evaluation. In conclusion, new methods of generating protons have the potential to be used in research of healthy tissue sparing as well as induction of immunologically useful tumor cells damage, which has chance to enhance the cancer treatment radiotherapy results.
Keywords: three-dimensional cell models, human colorectal carcinoma, laser-driven proton beam.
Acknowledgments: Institutional Grant RVO 61388971 (CZ), The National Institute for Cancer Research Project (EXCELES, ID Project No. LX22NPO5102) funded by the European Union - Next Generation EU, the Ministry of Health of the Czech Republic project NU23-08-00071, grant ELIUPM4-117, EU Marie Skłodowska-Curie Actions MERIT - Grant Agreement No. 101081195, the Generali/Česká pojišťovna a.s. (CZ), and UniCredit Bank CZ&SK a.s. Praha (CZ) sponsorships.
