NASA Technical Reports Server (NTRS) 20170000926: Skin-Based DNA Repair Phenotype for Cancer Risk from GCR in Genetically Diverse Populations

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SKIN-BASED DNA REPAIR PHENOTYPE FOR
CANCER RISK FROM GCR IN GENETICALLY
DIVERSE POPULATIONS
Elodie Guiet1, Louise Viger1,2, Charlotte Degorre2, Antoine Snijders1 , Sylvain V. Costes1,3
1Lawrence Berkeley National Laboratory, 2. University of Nantes, France, 3. NASA Ames Research Center
[email protected]
Background issue with non-confluent
Problem The Model
plates
Predicting cancer risk associated with cosmic
radiation remains a mission-critical challenge for
NASA radiation health scientists and mission
planners. Epidemiological data are lacking and
risk methods do not take individual radiation
sensitivity into account.
Hypothesis
Strain Radiation Sensitivity
In our approach we hypothesize that genetic factors strongly influence
risk of cancer from space radiation and that biomarkers reflecting DNA
POWER COEFFICIENT ALPHA FOR REPAIR
damage and cell death are ideal tools to predict risk and monitor
Power - high LET Power - Xray
Slope = Cst x hour^(-alpha(strain))
potential health effects post-flight.
CC019 CC037 CC032 CC051 CC011 B6C3 CC040 CC013 CC002 CC061 CBA C57 CC042 C3H BALBC
0
-0.2
Time and Dose points
-0.4
-0.6
-0.8
Year 1 – Project Overview -1
BACKGROUND (FOCI/CELL)
4
Bgd 350 MeV/n Si Bgd 350 MeV/n Ar Bgd 600 MeV/n Fe Bgd Xray
3
Strain 1 Strain 9 2
Strain 10
Strain 2
Strain 11
Strain 3
At this workshop, we will be reporting the work we have done over the Strain 4 Strain 12 1
Strain 13
Strain 5
Strain 14
Strain 6
first 9 months of this proposal. Skin cells from 15 different strains of
Strain 15 0
Strain 7
Strain 8 Empty CC019 CC037 CC032 CC051 CC011 B6C3 CC040 CC013 CC002 CC061 CBA C57 CC042 C3H BALBC
mice already characterized for radiation-induced cancer sensitivity
NORMALIZED SLOPE AT 48HRS (1/GY)
(B6C3F; BALB/cByJ, C57BL/6J, CBA/CaJ, C3H/HeMsNrsf), and 10 3
1 2 3 1 2 3
elam elam elam 1 ela 2 ela 3 ela elam elam elam 1 ela 2 ela 3 ela 2.5 Si_48hr Ar_48hr Fe_48hr Xray_48hr
e e e M M M e e e M M M
F F F F F F
strains from the DOE collaborative cross-mouse model were
2
1.5
expanded from ear biopsy and cultivated until Passage 3. On
1
average, 3 males and 3 females for each strain were expanded and 0.5
• Mouse skin cells exposed ex-vivo
0
• Duplicate plate per condition CC019 CC037 CC032 CC051 CC011 B6C3 CC040 CC013 CC002 CC061 CBA C57 CC042 C3H BALBC
frozen for further characterization at the NSRL beam line during the
Strains ranked from resistant to
NSRL16C run for three LET (350 MeV/n Si, 350 MeV/n Ar and 600
sensitive based on 48 hours
slope for X-ray
MeV/n Fe) and two ion fluences (1 and 3 particles per cell).
15 strains
X 3 males
4 8 24 48 144 Hours
X 3 females
Radiation
High-LET Control X2 LET
ASYMPTOTE AMPLITUDE FOR 4 HOUR XRAY FIT
L
N 1.1 Particles/100 µm2 X3 LET
7
B
3 Particles/100 µm2
6
Total
5
L X ray control
N Senescence
4
B X ray 0.1 Gy
L DSB (53bp1) 3
X ray 1 Gy
2
X ray 4 Gy
1
# of 96 well plates (DSB) 16+8 16 16+8 16+8 64+24 0
# of 96 well plates (Senescence) 16 16
Asymptotic fit
(Asym+(R0-Asym) x exp(-exp(lrc) x hour)
Linear fit
DSB/cell (8.75 DSB/Gy at 4 hours)
CORRELATION POWER FACTOR CORRELATION ASYMPTOTE VS REPAIR
-1 -0.9 -0.8 -0.7 -0.6 -0.5 2
Foci vs Track vs DSB
C
-0.2 IT
1.5
O
T T
EL -0.4 PM 1
- T y = 0.2033x + 0.5637
H Y I
G S F 0.5
A R² = 0.3267
I -0.6
H y = 1.1893x + 0.4455
Y
A 0
R² = 0.8531 R
-0.8 - 0 2 4 6 8
X
XRAY NORMALIZED FOCI HIGH-LET SLOPE AT 48 HOURS
Ranking metrics – Focusing on differences between low and high-LET
Experimental Design
Background Power high LET Power X-ray Xray4hr Asymptote High LET norm. slope 24/48 hr Xraynorm. slope 24/48 hr
CC019 0.63 CC019 -0.94 CC019 -0.69 CC019 1.81 CC032 0.69 CC019 0.32
CC040 0.64 CC051 -0.80 CC051 -0.51 CC051 2.56 CC011 0.90 CC037 0.45
BALBC 0.67 CC061 -0.77 CC040 -0.47 CC061 2.95 CC019 0.98 CC032 0.46
C3H 0.69 CC040 -0.76 CC013 -0.47 CC013 3.02 CC051 1.00 CC051 0.46
CC042 0.70 CC013 -0.75 B6C3 -0.43 CC040 3.14 CC061 1.07 CC011 0.47
B6C3 0.72 CC011 -0.73 CC011 -0.43 CC032 3.28 CC002 1.08 B6C3 0.49
Constants Variables Measurements
CC013 0.75 CC032 -0.72 CC061 -0.41 B6C3 3.58 BALBC 1.21 CC040 0.49
C57 0.79 BALBC -0.72 CC032 -0.40 CC011 3.60 CC037 1.30 CC013 0.52
CBA 0.82 CC042 -0.71 CC037 -0.40 C57 3.74 CC013 1.37 CC002 0.53
CC051 0.87 C57 -0.68 C57 -0.39 CC002 3.79 CC042 1.41 CC061 0.56
• Strains of • LET • 53BP1 foci detection as DNA
CC037 0.87 CC002 -0.67 CC042 -0.38 BALBC 3.98 CC040 1.63 CBA 0.61
CC061 0.92 B6C3 -0.67 BALBC -0.34 CC042 4.04 C57 1.67 C57 0.62
animals double strand break marker
• Dose
CC002 1.07 CC037 -0.66 CC002 -0.33 CC037 4.23 CBA 1.68 CC042 0.62
CC011 1.07 C3H -0.63 C3H -0.28 C3H 5.36 B6C3 1.76 C3H 0.74
• Human PBMC • Repair Kinetic parameters
• Time post- CC032 1.51 CBA -0.61 CBA -0.25 CBA 5.76 C3H 1.85 BALBC 0.75
(Year 2 and 3)
IR • (power function)
• Foci saturation (Asymptotic fit) Conclusion / Future Work
• Foci Background
• 800 cells/condition
The mice work has established new metrics for the usage of Radiation Induced Foci as a marker for various
53BP1 foci/cell
aspect of DNA repair deficiencies. In year 2, we propose to continue characterization of the mouse lines
with low LET to identify loci specific to high- versus low- LET and establish genetic linkage for the various
350 MeV/n Si
DNA repair biomarkers. Correlation with cancer risk from each animals strain and gender will also be
350 MeV/n Ar
investigated.
600 MeV/n Fe
On the human side, we will start characterizing the DNA damage response induced ex-vivo in 200 human’s
blood donors for radiation sensitivity with a tentative 500 donors by the end of this project. All ex-vivo
Linear fit (foci = Bgd + slope x hour) Tracks/cell DSB/cell (8.75 DSB/Gy at 4 hours)
phenotypic data will be correlated to genetic characterization of each individual human donors using SNP
arrays characterization as done for mice. Similarly, ex-vivo phenotypic features from mice will be associated
to cancer risk, to identify which biomarkers correlate the most with cancer risk. Genetic traits across
humans will also be associated to radiation phenotypic features as a function of age and gender.