The Laboratory of Radiation Biology Founders Vasilij Vasil'evich - PowerPoint PPT Presentation

The Laboratory of Radiation Biology

Founders Vasilij Vasil'evich Parin Oleg Georgievich Gazenko Andrej Vladimirovich Lebedinskij Jurij Grigor'evich Grigor'ev Page  2

First radiobiological experiments on synchrocyclotron Page  3

History 1959 First experiments at Laboratory of Nuclear Problems (LNP) 1978 Biological Research Sector at LNP 1988 Biological Department at LNP 1995 The Department of Radiation and Radiobiological Research 2005 Laboratory of Radiation Biology Prof. E.A. Krasavin, www.lrb.jinr.ru Corr.Member of RAS Page  4

Education activity The LRB performs education programs for students and young researchers on modern equipment Page  5

Main research topics 1. Research on the effect of accelerated heavy ions of different energies on genetic structures 2. Research on the effect of different doses of accelerated charged particles on the retina , study of cataractogenesis 3. Research on the character of the heavy charged particle-induced damage and functional disorders of central nervous system (CNS) cells. 4. Mathematical modeling of radiation induced effects in biophysical systems 5. Evaluation of the radiation environment and radiation safety 6. Solving problems of astrobiology ( in cooperation with Italy ) Page  6

JINR’s accelerators Phasotron: protons 660 MeV U-400M: heavy ions 50 MeV/u Nuclotron: heavy ions up to 4 GeV/u U-400: heavy ions 10 MeV/u Page  7

The dose distribution of radiation in matter 1 unit of the dose 1 unit of the dose X-rays Fe ion Page  8

The Galactic Cosmic Ray (GCR) energy spectrum The integral flux of GCR particles of carbon and iron groups equals to 10 5 particles/cm 2 per year Particle flux density interplanetary space Z ฀ 20 160 per day per cm 2 Page  9

Consequences of Galactic heavy ion action  Formation of gene and structural mutations;  Induction of cancer;  Violation of visual functions:  lesions of retina;  cataract induction;  CNS violation Page  10

DNA damage

Isolated DNA damage Single strand break Base damage Sugar damage Page  12

Clustered DNA damage Double strand break Base damage Sugar damage Base damage Page  13

“Comet assay” for detection of DNA lesions D = 0 Gy D = 5 Gy D =10 Gy mt Dose, Gy D = 60 Gy D = 20 Gy D = 40 Gy Page  14

Visualization of damaged sites in DNA Irradiation Fixation of cells at different times Secondary antibody post-irradiation (PI) with fluorescent dye Primary antibody Visualisation of   H2AX H2AX induced DSBs (γH2AX/53BP1 foci) 53ВР1 53ВР1 Acquisition of 53BP1 foci γH2AX foci images DSB DNA 3D analysis of induced γH2AX/53BP1 foci - Acquiarium Page  15 merge

Human cells exposed to γ -rays and 11 B ions 11 B ions (LET = 135 keV/μm) γ - rays (LET = 0.3 keV/μm) 5 min x-y y-z x-y y-z x-z x-z 1 h x-y y-z x-y y-z x-z x-z 2 μ m 24 h x-y y-z x-z x-y y-z x-z γ H2AX Page  16 53BP1 Chromatin (DAPI)

Human cells exposed to 11 B ions at 10 ° 5 min 15 min 30 min 4 h 24 h 1 h γ H2AX 53BP1 Chromatin (DAPI) Page  17

Kinetics of the formation and disappearance of γH2AX/53BP1 foci Comparison of γH2AX/53BP1 foci: γ -rays and 11 B Average number of γH2AX/53BP1 foci 80 γ -rays γ -rays 11 Boron 11Bor 60 per cell 40 20 0 Time after irradiation Page  18

Incidence of clusters of γ H2AX/53BP1 foci Comparison of total clusters: γ -rays and 11 B Average number of clusters per cell 20 11 Bor γ -rays 10 0 Time after irradiation Page  19

Monte-Carlo computer modeling of heavy ion tracks and cluster damage analysis GEANT4-DNA http://geant4.org Page  20

Mutagenesis and RBE

Radiation induced mutagenesis Structural mutation Gene mutation Guanine Page  22

The frequency of gene and structural mutation induction after γ -ray and heavy ion irradiation а б 4 He 50 4 He20 Nm/N  10 -5 Nm/N 12 C200 Dose, Gy Dose, Gy Page  23

Induction of mutagenic DNA repair by heavy ions luciferase FMNH 2 + RCHO + O 2  FMN + RCOOH +H 2 O + h  Page  24

RBE dependence on LET 2 3,2 3,0 3 2,8 2,6 2,4 1 1 – gene mutations R B E 2,2 x 2,0 2 – deletions 1,8 x 3 – lethal effect 1,6 x 1,4 x 1,2 x 1,0 0,8 0,1 1 10 100 1000 L E T , keV/  m Page  25

Formation of unstable chromosomal aberration in human cells after heavy ion irradiation Unstable chromosomal Aberrations/100 cells aberration Dose, Gy Block of cell division Page  26

Formation of stable chromosomal aberrations in human cells after heavy ion irradiation Chromosome № 1 Stable chromosomal Translocations/100 cells aberration Successful of cell division Dose, Gy Page  27

Cytogenetic effect of low doses of accelerated 24 Mg ions chromosomal aberrations The frequency of cells with 24 Mg chromosome aberrations. Chinese hamster cells exposed to 24 Mg γ -rays ions with energy 500 MeV/nucleon micronuclei Page  28

Genetic network model of induced mutagenesis in bacteria E.coli Page  29

Mathematical modeling of DNA repair systems in bacteria Page  30

Mathematical modeling of DNA repair systems in mammals and human Page  31

Action of radioprotectors

Influence of radioprotectors on bacterial cells after heavy ion irradiation without protector bacteria E.coli cysteamine LET, keV/mkm Page  33

Cancer

Gardner tumors H a r d e r ia n G la n d T u m o r P r e v a le n c e 3 0 G a m m a Iron ions p r o to n h e liu m n e o n R e la t iv e R is k ir o n ( 6 0 0 M e V /u ) ir o n ( 3 5 0 M e V /u ) 2 0 1 0  -rays 0 .0 0 .5 1 .0 1 .5 2 .0 D o s e , G y Page  35 Nelson, 2006

Skin cancer (rats) Page  36 Burns, Albert , 1986

RBE for carcinogenic effect of irradiation Page  37

Eyes and retina

Cataractogenesis UV-induced aggregation of Eye lens and retina  L - crystalline under B 11 cytoplasm micro-vacuolization, fiber electroretinogram cell swelling, nuclear fragmentation А Б МНМ, 70 mg/kg, 2h Control Dysfunction after mutagen insertion Page  39 Ostrovskii М.А., 2011

Cataract induction by iron ions and X-rays 10 1000 Iron ions Iron ions X-rays Cataract ratio 100 RBE 1 10 RBE = D x /D Fe 0.1 1 0.01 0.1 1 10 0.01 0.1 1 Dose, Gy Dose, Gy Worgul et al., 2006 Page  40

Action of 56 Fe ions on retina cells Axon growth index vs 56 Fe ion dose 90000 0 cGy 5 cGy 10 cGy 80000 * 70000 Arbitrary units 60000 50 cGy 100 cGy 200 cGy 50000 40000 30000 20000 0 50 100 150 200 250 dose, cGy Page  41 Apoptosis Vazquez, 2006

Central Nervous System

Cosmic ray hit frequencies in CNS critical areas Fe CNS in General  A B  2 or 13% cells will be hit at least one Fe p particle  8 or 46% would be hit by at least one p e particle with Z  15 p  Every nucleus will be traversed by a A B proton once every 3 days and a alpha particle once every 30 days. Mixed Field Multi-hit FE ION TRACKS VISUALIZED BY MARKERS OF DNA DSBs ( γ H2AX) 0 cGy 50 cGy 100 cGy 200 cGy TRACK DIRECTION

Cognitive tests (Morris water maze) 1 month after irradiation M.Rabin, 2005 Page  44

Persistent reduction in the spatial learning ability of rats after 56 Fe ion irradiation Results after 3 months 160 140 120 100 Delay, s 80 20 cGy 60 Ф  10 5 /cm 2 40 20 0 1 2 3 day of test R. Britten et al., 2012 Симуляция облучения 20 сГр 1 ГэВ/нуклон Fe 20 cGy 1GeV/nucleon 56 Fe control simulation Page  45

First experiments with monkeys Irradiation with a proton medical beam, 170 MeV Irradiation with 12 C ions, 500 MeV/u, at the Nuclotron Page  46

Distinguishing visual stimuli on the sensory attributes conditioned stimulus refreshment right response color brightness configuration orientation Page  47

Molecular targets at subcelular level Myelin sheath degradation Chiang et al (1993) RRP in glutamate synapses Levels of receptor subunits Shi et al (2006) Machida et al (2010) Britten et al (2014) Na + currents Mullin et al (1986); Hunt et al (1988); Sokolova et al (2015) Membrane hyperpolarization Sokolova et al (2015) Page  48

Rarefaction of Purkinje cell layer after irradiation by 645 М eV protons and 137 Cs  -rays Page  49 Krasavin Е.А., 1979

Studying the level of neurotransmitters in different rat brain areas Irradiation with 1 Gy of 500 MeV/u carbon ions Radiation-induced decrease in the level of neurotransmitters is observed in the brain regions responsible for the emotional and motivational state 3 months after irradiation Hippocampus Striatum Prefrontal cortex Nucleus accumbens Hypothalamus Page  50 Rat brain

Modeling of energy deposition events in CA1 pyramidal neurons Page  51

Modeling of cognitive tests by neural networks 20cGy 40cGy, Fe ions Simulation of single neuron and network activity during WM task Page  52