SECONDA CONFERENZA DEI PROGETTI DEL CENTRO FERMI 19-20 APRILE 2012

1. SECONDA CONFERENZA DEI PROGETTI DEL CENTRO FERMI 19-20 APRILE 2012 SALONE DELLE CONFERENZE DEL MINISTERO DELL’INTERNO ROMA, PIAZZA DEL VIMINALE 1 In vitro-In vivo MUrinecoSmiCsiLEnceexperiment (MUSCLE) D. Capece and E. Fratini (Centro Fermi) on behalf of the Cosmic Silence Collaboration

2. Aim of MUSCLE Project To understand the biologicaleffects of environmentalradiation

3. Peculiar low dose radiation effects (<100mGy) Phenomenon in which unirradiated cells exhibit irradiated effects as a result of signals received from nearby irradiated cells. • Bystander effect

4. Peculiar low dose radiation effects (<100mGy) Phenomenon characterized by an increase in the genetic spontaneous alteration observed in cells which have not themselves been irradiated, but are the progeny of an irradiated cell. • Genomic instability

5. Peculiar low dose radiation effects (<100mGy) Phenomenon that occurs when a low dose of mutagenic agent can protect from the biological damage of a subsequent high dose of the same or different mutagenica agents • Adaptive reponse Conditioning Dose Challenge Dose + Damage

6. Our Hypothesis Life has evolved on Earth for 3 bilion years in the presence of environmental ionizing radiation. All organism have therefore incorporated in their normal biology a daily stiimulus of ultra-low-dose environmental radiation that might act as conditioning agent.

7. LNGS – The Gran Sasso underground laboratory

8. Past Experiments Satta et al., 1995. Satta et al., 2002. Antonelli et al., 2008. S. Cerevisiae cells Carbone et al., 2009. Carbone & Pinto et al., 2010. Chinese Hamster cells Human cells

9. S. Cerevisiae D7 cells Satta et al., Mutat. Res. 1995.

10. Chinese Hamster V79 Satta et al., Radiat. Environ. Biophys 2002. Antonelli et al., Nuovo Cimento B 2008.

11. TK6 Lymphoblastoid cells Carbone et al., Radiat. Environ. Biophys 2009. Carbone & Pinto et al., Nuovo Cimento B 2010.

12. …Summarizing… Compatible with ourhypothesis of environmentalradiationas a conditioningagent, reducedenvironmentalradiationconditionsseem to render cells, afterseveralmonths of protracted culture: • Lesstolerant to radiation-induced DNA damage • Lessefficient in scavengingreactiveoxygenspecies

13. pKZ1 Mouse Model An ideal model for studying responses to very low doses of DNA damaging agents Matsuoka et al., Science 1991

14. pKZ1 Chromosomal Inversion Assay ZcaL P DNA Inversion LacZ P LacZ Expression Matsuoka et. al., Science 1991

15. Experimental design To use pKZ1 mouse chromosomalinversionassay to studybiologicaleffects of environmental background radiation • A11 Hybridoma cell line obtained from pKZ1 a splenocyte • Immortalized cell lines from brain and skin of neonatal pKZ1 mice Day et al., Radiat. Res. 1991

16. Experimental design Reference environmental radiation (~ 2mGy/year) Reduced environmental radiation (~ 25mGy/year) LNGS - INFN Istituto Superiore di Sanità Rome

17. Experimental design ISS – Reference Environmental Radiation LNGS – Reduced Environmental Radiation Rome Teramo Biological End-points • Inversion frequency • Doubling Time • Rate of apoptosis/autophagy • Redox cellular state

19. Perspectives Whathappensin vivo ? ReducedEnvironmentalRadiation LNGS (L’Aquila)/LSC-Canfranc underground laboratory(Saragozza)? Reference EnvironmentalRadiation L’Aquila University pKZ1 pKZ1

20. CosmicSilence Collaboration CENTRO FERMI D. Capece, E. Fratini, M.C. Carbone, M. Pinto INFN L. Satta Istituto Superiore di Sanità G. Simone, M.A. Tabocchini L’Aquila University F. Zazzeroni, E. Alesse ENEA Casaccia R. Amendola Flinders University, Adelaide P. Sykes ENEA R. Amendola INFN-LNGS M. Balata