OBJECTIVES:
This project addresses the need to develop and utilize mixed field irradiation protocols that approximately
represent the shielded tissue environment in space and that can be combined with single-beam studies to validate
and further improve cancer risk models. Extensive studies have been carried out on the induction of chromosomal
aberrations by low- and high- linear energy transfer (LET) radiation in human lymphocytes, fibroblasts, and
epithelial cells exposed in vitro. The results, which are consistent with computational modeling predictions,
establish chromosome aberration models as an LET-sensitive tool for predicting damage from single ion
exposures. However, there is a lack of data on chromosome aberrations induced by low dose rate chronic
exposure and mixed field beams such as those expected in space.
This investigation will define a simplified mixed field and make model predictions of the effects of dose rate,
mixed fields, and shielding on expected biological damage. Chromosome aberration studies at NASA’s Space
Radiation Laboratory (NSRL) will provide the biological validation needed to extend the computational models
over a broader range of experimental conditions leading up to the galactic cosmic radiation (GCR) simulator,
which will help to reduce uncertainties in radiation quality effects and dose-rate dependence in cancer risk
models. These models can then be used to answer some of the open questions regarding requirements for a full
GCR reference field, including particle type and number, energy, dose rate, and delivery order.
This study has the following specific aims:
1) Evaluate the effects of dose rate on biological damage using acute and chronic exposures with single ion
and mixed field beams to irradiate normal human fibroblasts and to measure chromosome aberrations.
2) Compare the external field versus local tissue field approach by measuring chromosome aberrations in
normal human fibroblasts using shielded and unshielded.
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APPROACH:
hTERT-immortalized human normal skin fibroblast cells 82-6 were grown as monolayers at under typical culture
conditions supplemented with 10% fetal bovine serum (FBS), antibiotic antimycotic and L-glutamine. Cells were
irradiated in a confluent state. The premature chromosome condensation (PCC) technique will be used to collect
G2/M-phase chromosomes. After irradiation, fibroblast were allowed to repair for 16 hours, and then subcultured
at low density. After 10-18 hours of incubation, cells were arrested in mitosis by adding colcemid to a final
concentration of 0.1 ug/ml in the culture media and then cells were incubated for an additional six hours.
Approximately 30 minutes before collection, 50 nM of calyculin were added to the culture media to condense the
chromosomes in the G2 phase of the cell cycle.
Whole blood was collected from healthy volunteers and irradiated with accelerated heavy ions. Immediately after
exposure, whole blood cultures containing RPMI 1640 media supplemented with 20% calf serum, and 1% were
incubated at 37 degrees Celsius.
Chromosomes were analyzed using the 3-color whole-chromosome fluorescence in situ hybridization (FISH)
technique during the first cell division after irradiation, and chromosome aberrations will be identified as either
simple exchanges (translocations and dicentrics) or complex exchanges (involving at least two chromosomes and
at least three double strand breaks).
RESULTS:
Chromosome analysis of 150 samples (human fibroblasts, epithelial cells and lymphocytes) collected during the
first three years were completed. Chromosome samples collected from mouse bone marrow 16 months after
exposure to single or mixed beams were analyzed. Both 4 beam mix and 6 beam mixed exposure shows less than
additive effects in chromosome aberration frequency both in human fibroblasts and blood lymphocytes. In
addition, 4 beam mix and 6 beam mix showed similar frequency of chromosome aberrations.
Crew health and performance is critical to successful human exploration beyond low Earth orbit.
The Human Research Program (HRP) investigates and mitigates the highest risks to human health
and performance, providing essential countermeasures and technologies for human space exploration.
Risks include physiological and performance effects from hazards such as radiation, altered gravity,
and hostile environments, as well as unique challenges in medical support, human factors,
and behavioral health support. The HRP utilizes an Integrated Research Plan (IRP) to identify
the approach and research activities planned to address these risks, which are assigned to specific
Elements within the program. The Human Research Roadmap is the web-based tool for communicating the IRP content.
The Human Research Roadmap is located at:
https://humanresearchroadmap.nasa.gov/
+ Click here for information of how this experiment is contributing to the HRP's path for risk reduction.