Short Title: Gender Differences in Radiation-induced Lung Adenocarcinoma

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:
Specific Aim 1: Determine whether there are sex differences in the incidence and
aggressiveness of lung adenocarcinomas after charged particle irradiation.

Aim 1A: Determine the incidence of lung cancer resulting from low LET radiation exposures
(137Cs [?-ray, 0.02 keV/?m]and proton exposures mimicking the 1972 Solar particle event [30-
180 MeV/n, 1.2-0.2 keV/μM]), and high LET exposures (28Si [600 MeV/n, 50.4 keV/μm] and 40Ca
[533 MeV/n, 107.7 keV/μm]) in male vs female BALB/c mice.

Aim 1B: Determine if there is a sex difference in the incidence of lung tumor aggressiveness
(invasion/metastasis) in the same irradiated cohorts in Aim 1a by pathologic interrogation.
Specific Aim 2: Characterize the lung tumors generated either spontaneously or by radiation
exposure at both the tissue and molecular level.

Specific Aim 2: Characterize the lung tumors generated either spontaneously or by radiation
exposure at both the tissue and molecular level.

Aim 2A: Determine if there are sex differences in the characterization of such tumors that may
point to underlying mechanisms for risk for carcinogenesis or aggressiveness of tumors via
transcriptomics and miRNA expression analysis of tumors. Confirm where possible the
molecular markers identified at the tissue level by histology and/or immune histochemistry.

Aim 2B: Determine if biomarker(s) based upon the appearance of circulating miRNA in plasma
or whether there are DNA copy number changes contained within circulating fragmented DNA
(cfDNA) that occur over time post-irradiation that are indicative of lung cancer risk from either
low vs. high LET radiation exposure.

Aim 2C: Determine if the appearance and extent of phosphotidyl serine on the exosomes found
in the blood of irradiated animals are a marker for lung cancer risk when examined over time
post-irradiation.

Specific Aim 3: Determine whether the FDA investigational new drug GC4419 can mitigate
spontaneous and radiation-induced lung cancer incidence in animals when exposed to
simulated GCR and SPE’s.

Aim 3A: Determine the incidence of lung cancer in mice when GC4419 is provided ad libitum
just before and for 20 weeks post-irradiation.

Aim 3B: Determine whether there are molecular changes (transcriptomics and miRNA
expression) in the lung tissues of irradiated animals without tumors as a function of time when
exposed to GC4419 or not vs unirradiated controls.

Aim 3C: Using the molecular signatures established in Aim 2, determine if GC4419 alters such
signatures that were indicative of either high vs. low LET radiation or tumor aggressiveness and
whether there is a correlation between GC4419 exposure, lung cancer incidence, and the
appearance of circulating, exosomal phosphotidyl serine.

Short Title: GCR, DNA repair and countermeasures

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:

Aim 1. Removed from study.

Aim 2. To test the hypothesis that GCR exposure leads to a chronic DNA damage response in mouse lung and whether this can be abated with a dietary countermeasure, nicotinamide riboside. Sex, dose and radiation quality effects will be investigated.

Aim 3. To determine whether radiogenic tumors display a mutational signature indicative of a decline in the functional status of the DNA repair machinery as reflected in their mutational signature, thus establishing a functional link to tumorigenesis. This aim has been expanded from the original study to include a pilot experiment to determine fixed-dose sex-specific radiation effect ratios (RERs) for lung tumorigenesis. Sex, radiation quality, and countermeasure effects will be investigated.

Short Title: Mouse Glioma Models

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:

Aim 1: Assess radiation quality effects on charged particle-induced carcinogenesis using validated

glioblastoma models.

Aim 2: Delineate the role of NHEJ vs HR DNA repair pathways as barriers to charged particle-induced gliomagenesis.

 

Aim 3: Identify genetic signatures unique to charged particle-induced glioblastomas.

Short Title: Mouse Models Cancer Risk & Prevention

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:

Aim 1: Investigate the cellular and molecular mechanisms that contribute to the space radiation associated carcinogenic risks of mouse models of human colon and lung cancer.

 

Aim 2: Determine the probabilities for increased colonic and lung carcinogenesis from space radiation as a result of radiation dose, quality, and dose rate and to compare the biomarkers obtained from these cancer susceptible mice with those in wild type mice so we can extrapolate to risks in healthy astronauts.

 

Aim 3:  Provide statistically significant biomarker signatures and biological markers

for modeling and risk assessment for increases in cancer for astronauts during long-term space

missions.

 

Aim 4: Determine the anti-cancer effectiveness and long-term side effects, if any, of the CDDO

countermeasure to guide the most effective approaches for their use on long-term space missions.

Short Title: MERA

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Directed

Aims:

The Multi-model Ensemble Risk Assessment project will develop an ensemble modeling approach for risk assessment where REID acceptance is determined based on the region of overlap between multiple predictive models allowing a straight forward method to input information derived from multiple sources including international risk prediction models.

 

Specific Aims:

Aim 1:  Determine mathematical formulism to implement multiple probabilistic and deterministic risk models into a single risk framework.

Aim 2: Identify common risk quantification parameters across cancer, cardiovascular disease, and late neurodegeneration consistent with Human System Risk Board quality of life consequences.

Aim 3: Develop visualization of results to support risk communication and decision making.

Aim 4: Develop multi-scale model for the evaluation of countermeasure efficacy based on human epidemiology and radiobiology data from NSRL.

Aim 5: Develop methodology to down select models for inclusion and retirement including appropriate weighting factors.

 

Models will be configuration managed and fully documented in accordance with appropriate NASA software develop standards

Short Title: Ovarian cancer and space radiation

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:
Specific Aim 1: Utilize archived ovaries to compare ovarian tumor induction by irradiation with oxygen (16O), iron (56Fe), or g-rays and investigate the mouse strain dependence of ovarian tumor induction by 16O and 56Fe. Comparison of ovarian tumorigenesis in 3-month old C57BL/6J and B6C3F1 mice irradiated with 0.5 Gy 16O or 56Fe with 3-4 month old CB6F1 mice irradiated with 0.4, 0.8, 1.2, or 1.6 Gy g-rays, and concurrent controls, all euthanized 15-16 months after irradiation. We will conduct detailed histopathology of ovaries and molecular characterization of tumors using immunostaining for tumor markers. We will establish the relative biological effectiveness of charged 16O or 56Fe compared to g-radiation for ovarian tumorigenesis.

Specific Aim 2: Utilize archived ovaries harvested at various time points after irradiation
with low doses of 16O or 56Fe to examine the persistence of ovarian oxidative lipid, protein,
and DNA damage, archived serum to measure a biomarker of ovarian reserve, and evaluate
these as potential early biomarkers of ovarian tumorigenesis. Our published work [Mishra et
al. 2016, Mishra et al. 2017] demonstrates oxidative damage and dose-dependent apoptotic
depletion of ovarian follicles after exposure to 0, 0.05, 0.3, and 0.5 Gy 16O or 56Fe, with estimated
ED50 for follicle depletion at one week after irradiation of 0.046 and 0.27 Gy, respectively. Serum
luteinizing hormone (LH) and follicle stimulating hormone (FSH) were significantly elevated in
0.5 Gy-irradiated mice 8wk after irradiation, consistent with loss of negative feedback due to
follicle depletion, but serum LH and FSH are not optimal serum markers of ovarian reserve
because they vary with estrous cycle stage and are secreted episodically, and we did not perform
immunohistochemical analyses of ovaries at 8 wk after irradiation. We will examine oxidative
lipid, protein, and DNA damage by immunostaining as potential biomarkers of ovarian tumor risk
in archived ovaries from mice euthanized 8 wk after irradiation with 16O and 56Fe. We will measure
Anti-Müllerian Hormone (AMH), a serum marker of ovarian reserve that is used clinically, in
archived serum from mice euthanized 1 wk and 15 months after irradiation.

Short Title: Gastrointestinal Cancer

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:

Aim 1: Quantitatively assess GI tumorigenesis in mouse models of GI cancer and collect samples for qualitative analysis.

 

Aim 2: Dissection of the signaling events and consequences in GI cells of the persistent effects of space radiation (e.g. oxidative stress, aberrant pro-proliferative signaling, and ongoing mitochondrial damage and dysfunction)

 

Aim 3: Characterization of IR-induced neoplastic events in normal diploid human colonocytes. In order to bridge in vivo mouse model tumorigenesis studies, human colonocytes will be employed to characterize space radiation induced changes and to compare to colonocytes from irradiated mice (Aims 1 and 2).

 

Aim 4: Development of systems biology (Project 4A) and mathematical modeling (Project 4B) approaches for GI cancer risk assessment. Genome-wide NGS combined with other omics approaches will contribute to a systems biology understanding of the effects of space radiation in the intestines and stomach.

Short Title: Simulated Space Radiation Interactions

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Human Health Countermeasures (HHC) Element

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:
Aim 1: Examine the metabolome of mice exposed to space flight relevant radiation.

Aim 2: Determine changes in the GI associated immune system.

Aim 3: Measure changes in the architecture of the GI associated lymphoid organs and lymphatic vessels of the bowel wall.

Short Title: Cancer Biomarkers

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:

Project 1 - identify biomarkers for susceptibility to HZE ion carcinogenesis and for early disease detection using genetic and systems biology approaches. Uses integrative “omics” approaches over multiple levels of biological organization and involves multiple species.

Project 2 - focuses on the mechanisms underlying the increased malignancy of HZE ion-induced tumors compared with their spontaneous or γ- ray-induced counterparts.

Project 3 - examines the critical question of risk from protracted exposures to high LET radiation at low doses and dose rates. Uses chronic exposures to high LET associated neutron radiation as a surrogate for conditions of space-relevant fluence rates and total doses to estimate the carcinogenic effects.

Project 4 - utilizes the resources (irradiated mice and “omics” results) generated in the first three projects to study the neurobehavioral consequences of HZE ion and neutron exposures and whether they are related to cancer outcome measures and predicted by the same or distinct biomarkers

Short Title: Epigenetic state modulation of radiation-induced DNA damage

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:

Aim 1: Build coarse-grained Monte Carlo simulations to sample chromatin fiber ensembles for selected epigenetic states based on data from human fibroblasts.

Aim 2. Simulate irradiation of chromatin fiber models and predict DNA break patterns.

Aim 3. Predict and DNA fragmentation patterns in different epigenetic states in IMR-90 fibroblasts and HUVECs after irradiation with a galactic cosmic ray model and X-rays.

Short Title: Space radiation effects on clonal hematopoiesis

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:
Aim 1: Employ a novel murine adoptive bone marrow transfer approach to model clonal hematopoiesis induced by somatic mutations in TET2, DNMT3A, TP53 and PPM1D. 

Aim 2: Examine the consequences of simulated GCR and SPE on: (1) hematologic clone expansion (2) cardiovascular function (3) leukemogenic/cancer endpoints. 

Short Title: Chromosome Aberrations II - Breakpoints

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Directed

Aims:

Aim 1: determine if analysis of IR-induced CA breakpoint junctions suggest different molecular pathways are involved, with respect high- versus low LET exposure.

Aim 2: determine if the sizes of co-deleted sequences at breakpoint junctions differ for those produced by high versus low LET IR; these may form the basis of an LET-dependent biomarker.

Aim 3: determine if the breakpoints associated with complex chromosomal exchanges are different than those associated with simple reciprocal exchanges that involve only two chromosomes.

Aim 4: At NASA’s discretion related to source availability and energies of interest, examine the breakpoint junctions of exchange aberrations induced by medium-to-high energy neutrons.

Aim 5:  At NASA’s discretion, characterize molecular differences between the breakpoints produced by different ultra-low dose ions.

Short Title: GCR Harderian Gland

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Active - Currently funded and in progress

Procurement Mechanism(s):

Solicited

Aims:

Aim 1: Investigate the effects of a single beam 350 MeV/u Oxygen (LET ~ 20 keV/μm) alone at 10, 20 and 30% tumor frequency levels and two-beam combinations of (1) 250 MeV/u protons (LET ~ 0.4 keV/μ) + 260 MeV/u Silicon (LET ~ 70 keV/μm) and (2) 250 MeV/u protons+600 MeV/u Iron (LET ~ 190 keV/μm) and (3) 260 MeV/u Silicon + 600 MeV/u Iron on the development of HG and lung tumors 16-month post irradiation. The investigator will measure the tumor prevalence in B6CF1/Anl mice whole-body exposed. In addition to the sham-treated controls, a minimum of 3 dose groups with protons contributing to 60% of the dose and 40% of the dose for Z>2.

 

Aim 2: Carry out corresponding multi-component experiments, with the same beam energies as in Aim 1 with at least 3 dose groups in addition to the sham treated controls.

 

Aim 3: In silico modeling of mixed-beam and other experiments on tumor incidence.

Short Title: MERA Maturation

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Planned-Funded - Task expected to be within budget

Procurement Mechanism(s):

Maturation

Aims:

Ongoing use of ensemble modeling approach for risk assessment where REID acceptance is determined based on the region of overlap between multiple predictive models to integrate: 

1. Common risk quantification parameters across cancer, cardiovascular disease, and late
    neurodegeneration consistent with Human System Risk Board quality of life consequences.
2. Countermeasure efficacy based on human epidemiology and radiobiology data from NSRL.

Short Title: Dose Rate Effects and GCR Simulation

Responsible HRP Element: Space Radiation

Collaborating Org(s):

Funding Status: Planned-Funded - Task expected to be within budget

Procurement Mechanism(s):

Directed

Aims:

• Test the hypothesis that the time between doses and the dose-rate are critical modulators of mutant frequency in human cells following exposure to a mixed GCR field. Cultured human TK6 lymphoblasts will be used and the results compared with data obtained previously with these cells using acute exposures to single ions or rapidly delivered sequential beams.

• Test the hypothesis that the time between doses and dose-rate are critical modulators of mutational spectra following exposure to a mixed GCR field. We hypothesize that the types of TK1 mutations will be distinct from spontaneous events and that dose-rate and the time between exposures will alter the signature of radiogenic mutations.