ELI-RO 10/2024 Project
English (Engleză)
Project title: Understanding FLASH irradiation effect on cancer cells using laser-induced very high energy electrons and in vitro tumor-on-chip assays / FLASH CANCELLS
Periode: 11 July 2024 – 31 March 2027
Contact Person:
Project Director: Dr. Felix SIMA (INFLPR)
Coordinator:
National Institute for Laser, Plasma and Radiation Physics - I.N.F.L.P.R.
Atomiștilor 409, Măgurele, Romania
Partner:
Institutul de Biochimie al Academiei Romane (IB-AR)
Splaiul Independentei, nr. 296, sector 6, 060031, Bucharest
Abstract (Objectives):
The main objective of this project is to propose and exploit various innovative in vitro approaches in order to understand the FLASH effect induced by either laser-plasma accelerated very high energy electrons (LPA VHEE) or X-ray radiation on cancer cells. To this aim, we will investigate radiation dose dependent cancer cell death vs. potential immune boosting properties, using a panel of chemo- and radio- sensitive or resistant cancer cells together with immune and tissue-specific non-malignant normal cells. Specifically, the project aims to increase the understanding of molecular mechanisms involved in cellular response to FLASH radiation by providing fundamental knowledge of its effects using in vitro models of melanoma and ovarian cancer. A tumour-on-chip model will be proposed for testing the immune response upon co-culture of lymphocytes with cancer spheroids. The tumour-on-chip concept will be designed to test the interaction of cancer cells with artificial narrow tissue-like architectures and immune system components and live monitored by time lapse microscopy in parallel for cancer cells exposed to FLASH irradiation vs. unexposed cancer cells. Other specific objectives refer to i) the development of an X-ray FLASH experimental demonstrator for ultra-high dose rate delivery exceeding 40 Gy/s, ii) the improvement of LPA VHEE experiment reliability by efficient control of laser peak intensity and focus position iii) the development of an LPA geometric configuration to obtain VHEE beams with very high charge and iv) the development of new dosimetric characterization protocols.
Phases:
- 2024 - Analysis of cancer cell response upon exposure to pulsed X-ray and conventional X-ray irradiation conditions;
- 2025 - Analysis of cancer cell response upon exposure to FLASH X-ray irradiation conditions;
- 2026 - Analysis of cancer cell response upon exposure to LPA VHEE for FLASH irradiation and potential synergy with immunotherapeutic (biological) agents;
- 2027 - Validation of proteome analysis identified cancer cell secreted biomarkers of radiotherapeutic response using cell biology and biochemical assays.
Română (Romanian)
Titlul proiectului: Intelegerea efectului de iradiere FLASH asupra celulelor canceroase utilizand fascicule de electroni de energie foarte mare accelerati cu laser si analize in vitro tumora-pe-cip / FLASH CANCELLS
Perioada: 11 Iulie 2024 – 31 Martie 2027
Persoana de contact:
Director Proiect: Dr. Felix SIMA (INFLPR)
Coordonator:
Institutul Național pentru Fizica Laserilor, Plasmei și Radiației - I.N.F.L.P.R.
Atomiștilor 409, Măgurele, ROMÂNIA
Partener:
Institutul de Biochimie al Academiei Romane (IB-AR)
Splaiul Independentei, nr. 296, sector 6, 060031, Bucuresti
Abstract (Obiective):
Obiectivele principale ale acestui proiect sunt propunerea și exploatarea diferitelor abordări in vitro cu scopul de a înțelege efectul FLASH indus asupra celulelor canceroase atât de electronii de energii foarte înalte (very high energy electrons – VHEE) generați prin accelerare laser-plasma (laser-plasma accelerated – LPA), cât și de radiația X. Astfel, propunem investigarea mecanismelor de apoptoză celulară în funcție de doza de iradiere vs. potențiale proprietăți de stimulare imună, folosind celule canceroase rezistente sau sensibile radiologic și chimic, împreună cu celule imune si celule non-maligne sănătoase, specifice tesutului de origine. În mod specific, proiectul propune înțelegerea mecanismelor moleculare implicate în răspunsul celular la radiație FLASH prin furnizarea de cunoștințe fundamentale asupra efectului acesteia folosind modele in vitro de cancer de melanom și cancer ovarian. Un model tumoral pe biocip va fi propus pentru testarea răspunsului imun pe modele de co-cultură de limfocite cu sferoizi tumorali. Conceptul de model tumoral pe biocip va fi elaborat pentru testarea interacției celulelor canceroase cu arhitecturi artificiale constrictive ce mimează țesuturile și cu componente ale sistemului imun si va fi monitorizat prin microscopie în timp real, atât în cazul celulelor expuse la iradieri FLASH, cât și în cazul celulelor neexpuse. Alte obiective specifice se referă la i) dezvoltarea unui demonstrator experimental de raze X de tip FLASH pentru doze ultra-înalte de peste 40 Gy/s, ii) îmbunătățirea fiabilității experimentelor bazate pe VHEE generați prin LPA prin controlul precis al intensității de vârf a laserului și poziția de focus a acestuia, iii) dezvoltarea unei configurații geometrice LPA pentru obținerea fasciculelor VHEE de sarcină foarte înaltă și iv) dezvoltarea de noi protocoale de caracterizare dozimetrică.
Etape:
- 2024 - Analiza răspunsului celulelor canceroase dupa expunerea la diverse condiții de radiație X pulsată și convențională;
- 2025 - Analiza răspunsului celulelor canceroase dupa expunerea la diverse condiții de radiație X de tip FLASH;
- 2026 - Analiza răspunsului celulelor canceroase la expunerea cu electroni VHEE LPA pentru iradieri FLASH și a potențialei sinergii cu agenți imunoterapeutici (biologici);
- 2027 - Validarea analizei proteomice realizate pe biomarkeri secretați de celule canceroase ca răspuns la radioterapie folosind experimente de biologie celulară și biochimie.
RESULTS:
Publications:
[1] S. Orobeti, I. Porosnicu, A. Bran, I. Tiseanu, F. Sima, S. M. Petrescu, and L. E. Sima, "In vitro experiments with cell co-cultures exposed to LPA electrons generated by the interaction of a high-intensity laser beam with gas jets", submitted to Bio-protocol, October 2024.
[2] A. Bran, S. Orobeti, F. Jipa, A. Bonciu, E. Axente, L. E. Sima, F. Sima, K. Sugioka, Tissue-like scaffolds created by two photon polymerization for testing cancer cell behavior in confined environments, submitted, November 2024.
[3] L. E. Sima, M. Tudor, S. Orobeti, A. Filimon, G. Negroiu, C. Munteanu, G. Chiritoiu, A. Roseanu, P. Florian, M. Icriverzi, I. Porosnicu, D. Avram, A. Bran, S. Simion, I.Tiseanu, F. Sima, Ultra-high dose rate ionizing radiation for FLASH radiotherapy applications, Chapter in Book "Ionizing Radiation - Insights into Chemistry, Biology, and Applications", Ed. Intech, submitted, December 2024.
Patents:
Conference presentations:
[1] L. Ionel, F. Jipa, A. Bran, E. Axente, G. Popescu-Pelin, F. Sima, K. Sugioka, Numerical and experimental investigations of ultra-short pulsed laser interaction with matter in focal region for advanced lab-on-chip applications, 16th International Conference on Physics of Advanced Materials (ICPAM-16), Invited presentation, Akdeniz University in Antalya, Turkey, September 7-15, 2024.[2] L.E. Sima, S. Orobeti, I. Porosnicu, C. Diplasu, G. Giubega, G. Cojocaru, R. Ungureanu, C. Dobrea, M. Serbanescu, A. Mihalcea, E. Stancu, C. E. Staicu, F. Jipa, A. Bran, E. Axente, S. Sandel, M. Zamfirescu, I. Tiseanu, F. Sima, First molecular investigations using laser-induced high-energy electron FLASH irradiation to characterize cancer cells response to ultrahigh dose rate exposure, The Annual International Conference of the RSBMB 2024, Keynote presentation, Romanian Academy Library, Bucharest, Romania, September 25-27, 2024.
[3] A. Bran, S. Orobeti, L.E. Sima, F. Jipa, A. Bonciu, E. Axente, K. Sugioka, F. Sima, 3D tissue-like scaffolds fabricated by two-photon polymerization for melanoma cell invasion evaluation, The Annual International Conference of the RSBMB 2024, Oral presentation, Romanian Academy Library, Bucharest, Romania, September 25-27, 2024.
[4] S. Orobeti, G. Chirițoiu, M. Tudor, A. Bran, F. Sima, S. Petrescu, L.E. Sima, SKOV3 actin-GFP ovarian cancer stable cell lines for 4D imagistic analysis of cancer cell processes upon treatment with chemo- and FLASH radiotherapy, The Annual International Conference of the RSBMB 2024, Poster presentation, Romanian Academy Library, Bucharest, Romania, September 25-27, 2024.
[5] F. Sima, A. Bran, F. Jipa, L. Ionel, S. Orobeti, E. Axente, L. E. Sima, K. Obata, M. Hanzawa, H. Kawano, K. Sugioka, Tumor-on-chip models for testing cancer cell behavior in tridimensional environments, Congresul Universităţii de Medicină și Farmacie „Carol Davila” din București, XII-a ediție, poster and oral presentation, 24-26 October 2024, Bucuresti, Romania
[6] M. Tudor, C. Munteanu, G. Chiritoiu, S. Orobeti, F. Jipa, F. Sima, G. E. Schiltz, D. Matei, L. Sima, Small molecule inhibitors targeting TG2 in ovarian cancer and identification of drug escape pathways, Congresul Universităţii de Medicină și Farmacie „Carol Davila” din București, XII-a ediție, poster and oral presentation, 24-26 October 2024, Bucuresti, Romania.
[7] F. Sima, A. Bran, K. Obata, M. Hanzawa, H. Kawano, F. Jipa, L. Ionel, S. Orobeti, L. E. Sima, E. Axente, K. Sugioka, Ultrafast laser 3D processing of tumor-on-chip platforms for cancer research, Invited presentation, 5th International Conference on Optics, Photonics and Lasers, 25-27 November, Valencia, Spain.
[8] A. Bran, F. Jipa, A. Bonciu, S. Orobeti, E. Axente, L. E. Sima, F. Sima, K. Sugioka, Cancer cells interaction with 3D tissue-like scaffolds fabricated by two photon polymerization, Oral presentation, 5th International Conference on Optics, Photonics and Lasers, 25-27 November, Valencia, Spain.
[9] L.E. Sima, S. Orobeti#, I. Porosnicu, C. Diplasu, G. Giubega, G. Cojocaru, R. Ungureanu, C. Dobrea, M. Serbanescu, A. Mihalcea, E. Stancu, C.E. Staicu, F. Jipa, A. Bran, E. Axente, S. Sandel, M. Zamfirescu, I. Tiseanu, F. Sima, First in vitro tests of laser-induced very high energy electron FLASH irradiation effect on cancer cells, Invited presentation, OncoHub 2024, November 21-23, 2024, Hotel Pullman, Bucharest, Romania.
[10] S. Orobeti, G. Chiritoiu, M. Tudor, A. Bran, F. Sima, S. Petrescu, L.E. Sima, Developing in vitro models for real-time imaging analysis of cancer cell processes in response to chemotherapeutics and radiation exposure, Poster presentation, 4th edition of OncoHub 2024, November 21-23, 2024, Hotel Pullman, Bucharest, Romania.
Webinars:
[1] L.E. Sima, Quantification of p-𝛾-H2AX foci in co-cultured cells exposed to radiation, invited Zoom recorded webinar (with Karly Buffa, TissueGnostics, Austria), released on November 2024, (https://tissuegnostics.com/webinar-livia-sima).
Outreach:
[1] L.E. Sima, participating in AFIST 2024 – topic: „Passion and progress. Women who inspire STEM fields”, November 27, 2024, organized by the Association of Women in Engineering, Science, and Technology (AFIST) under the auspices of CNR UNESCO, Invited interview, House of Trade and Industry, Bucharest (CCIB), Romania.
Trainings:
[1] Alexandra Bran, Working stage at Center for Advanced Photonics, RIKEN, 4-27 October, RIKEN, Wako, Japan; Felix Sima, Working visit at Center for Advanced Photonics, RIKEN, 4-14 October, RIKEN, Wako, Japan
[2] Ioana Porosnicu, The XII Geant4 International School, 25-29 November 2024, ELI-NP, Magurele, Romania
Awards:
International Cooperation:
[1] RIKEN, Center for Advanced Laser Photonics, Japan, Professor Koji Sugioka;