Aim 1: To determine the effects of X-ray radiation on RAS, pro-oxidant and pro-fibrotic markers in the heart. The proposal hypothesizes that the combination of fish oil with pectin will mitigate radiation-induced pathology.

Aim 2: The impact of HZE (28Si, 48Ti) radiation on cardiac markers of pro-oxidant and pro-fibrotic signaling will be determined. This proposal postulates that intervention with fish oil and pectin will abrogate radiation-induced oxidative stress and fibrosis in the heart.

Aim 3: The effect of HZE (56Fe) radiation combined with X-Ray radiation at BNL will be determined on RAS, oxidative stress, pro-inflammatory markers, and pro-fibrotic signaling on heart. The proposal hypothesizes that deletion of a single p53 allele will significantly affect markers of oxidative stress, inflammation, and fibrosis.

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.

• Aim 1: Identify clinical characteristics, biofluid biomarkers and imaging features on baseline and follow-up multiparametric MRI that predict for cognitive decline following radiation exposure to the brain.
• Aim 2: Evaluate the relationship between changes in specific aspects of cognitive function with radiation dosimetry in the whole brain and brain subregions in patients receiving radiation to the brain.
• Aim 3: Identify early changes in multiparametric MRI measures that predict for subsequent changes in brain structure following radiation to the brain in preclinical and clinical studies.

• Aim 1: investigate late CNS/CVD Effects of Simplified 5-ion Mixed Field Beam (normalized to 500 mGy) GCR IRR in Male & Female AD Tg and WT Mice.

• Aim 2: investigate Late CNS/CVD Effects of Simplified 5-ion Mixed Field Beam IRR in Male and Female APP/E3 and APP/E4 Tg and WT Mice.

• Aim 3: investigate Acute and Late CNS and CVD Effects of Simplified 5-ion Mixed Field Beam Space Radiation on Human Cells In Vitro

Aim 1: Perform comprehensive evaluation of physiological and functional changes occurring in the     

murine cardiovascular system following exposure to different HZE radiation scenarios.  

Aim 2: Correlate cardiovascular functional changes with molecular pathway perturbations obtained from proteomic data sets (quantitative, dynamic and post translational modification).

Aim 1: Determine the effects of chronic low-dose rate neutron exposure on cardiac function and structure in male C3H and female BALB/c mice.

Aim 2: Assess how Long Evans rats scale to C3H and BALB/c mice in their cardiovascular response to chronic low-dose rate neutron exposure.

Aim 3: Identify similarities and differences in the cardiovascular response to low dose protons and 16O exposure in the mouse, rat, and rabbit.

Aim 1: In the archived heart samples of fractionated proton and iron IR studies the study

plans to determine long-term IR-induced effects on cardiac fibrosis, inflammation, oxidative damage, microvascular, structural and morphological changes in the heart, including cellular hypertrophy and cytoskeletal reorganization.

Aim 2: In archived samples of CB6F1 mice irradiated with a range of low and very low doses of gamma, 14Si, 22Ti and 56Fe the study proposes to determine long-term effects and molecular endpoints in the heart as a function of IR type and dose.

AIM 1. Determine the longitudinal effect of IR type, dose and gender on cardiovascular physiology in wild type mice and isolated CV cells after single, low-dose, full-body proton (¹H), HZE particle iron (⁵⁶Fe) and mixed field and gamma radiation (? -IR).

AIM 2. Evaluate space-type IR mediated modulations in exosomal cargo in the blood, and determine whether these changes are associated with alterations in the heart function, structure and vasculature before manifestation of clinical symptoms.

AIM 3. Utilize known and newly identified biomarkers in the blood to develop human relevant point-of-care tests (POCT) for predicting and monitoring possible CV alterations before, during and after the space flights.

The objectives of this application are to characterize the impact of radiation doses and qualities on retinal vasculature and tissue remodeling and to study the role of mitochondria in regulating oxidative stress-induced cellular damage and alteration of cell-cell interactions in the function of blood-retina-barrier (BRB). Archived ocular tissues samples at dose ranges from 0.1 Gy to 1 Gy with multiple particle types and energies across multiple animal species will be available for sharing from three NASA funded investigators. We will evaluate radiation –induced retinal vascular and tissue remodeling at multiple time points. Our study will elucidate the spectrum of radiation-induced structural and functional alteration in retinal BRB function and the mechanisms by which these

changes are mediated.

Specific Aim 1:
Define the relationships between radiation-induced oxidative stress in ROS expression, retinal vascular remodeling, and BRB function. We will correlate cellular oxidative status with changes in retinal vascular topology, vascular endothelial cells (ECs), and BRB integrity using immunohistochemistry (IHC), stereological and automated image analyses, and magnetic resonance imaging diffusion tensor imaging (DTI-MRI). The observed changes in mice will be compared with that of rats and rabbits.

Specific Aim 2:
Determine whether radiation-induced oxidative damage in retina is mediated through photoreceptor mitochondrial ROS production. We will examine mitochondrial associated-oxidative stress and apoptosis and mitochondrial integrity in retinal photoreceptor using Western blot assays and histopathology.

Aim 1. To compare and contrast the efficacy of countermeasures to High-LET exposure

in cultured heart cells.

Aim 2: Compare and contrast the efficacy of druggable molecules in the long-term

management in an animal model of high-LET radiation induced cardiomyopathy.

Aim 1:  To histologically analyze cardiac morphology and cardiac oxidative stress of mice

exposed to space radiation.

Aim 2: To appraise the levels of mitochondrial oxidative stress in response to space radiation.

Aim 3. To determine the status of mitochondrial stress response pathways.

Aim 1. Create RBEs for the vascular damage endpoints.

• Aim 1a Carry out Gamma radiation studies at BNL for the determination of reference RBEs.
• Aim 1b Carry out charged particle studies at BNL for the determination of RBEs

Aim 2. Identify the proteins secreted by the endothelial cells during angiogenesis and in mature human 3D micro-vessel tissue models in response to radiation.

• Aim 2a Development of the proteomics assay.
• Aim 2b Identify the proteins secreted by the endothelial cells during angiogenesis and in mature human 3 D micro-vessel tissue models in response to gamma radiation.
• Aim 2c Identify the proteins secreted by the endothelial cells during angiogenesis and in mature human 3 D micro-vessel tissue models in response to the Simplified GCR sim.

• AIM 1: Discovery and validation of miRNAs biomarkers that are altered as a function HZE exposure, at a dose that could result in increased cardiovascular risks in astronauts on long duration space mission.

• AIM 2: Discovery and validation of miRNAs and lncRNAs altered as a function of exposure to 0.2 Gy Silicon ions in C3H mice and matched sham controls.

Aim 1. Establish temporal profiles of human telomere length dynamics (before, during, and

after flight) for individual astronauts across the integrated One-Year Mission Project onboard the

ISS, and across the concurrent ground analog component (prolonged isolation missions).

• Evaluate telomere length dynamics (qRT-PCR) and shifts in individual telomere length

distributions (Telo-FISH); bulk vs. stem/early progenitor cell populations

• Evaluate telomerase status/expression (TERT and HIF-1a protein levels; ELISA)

Aim 2. Establish temporal profiles of DNA damage responses (before, during, and after flight)

for individual astronauts across the integrated One-Year Mission Project onboard the ISS, and

across the concurrent ground analog component (prolonged isolation missions).

• Evaluate DNA damage responses; genomic vs. telomeric (g-H2AX foci, TIFs)

• Evaluate cytogenetic damage/biodosimetry; genome instability, telomere-specific

oxidative damage, and ALT activation (Telo-dGH/G-rich sister telomere loss/T-SCE)

Aim 1. Characterize and build computational models for the effects of age-at-exposure and dose

fractionation (repeated exposures) on performance in each of three behavioral domains, using

NASA-provided data sets.

Aim 1a. Evaluate the age-at-exposure and dose fractionation data sets for data quality across ions, exposure regimens, study design, and behavioral outcomes, and for suitability for multivariate modeling of risks for specific behavioral domains.

Aim 1b. Build single-ion risk models that incorporate age-at-exposure and dose-fractionation effects for each behavioral domain, using data sets that pass the Aim 1a evaluation criteria.

Aim 1c. Build multi-ion risk models that integrate the age-at-exposure and dose-fractionation effects for each behavioral domain from the single ion models of Aim 1b. The risk predictions of the different behavioral domains will be evaluated using both experimental exposure regimens and GCR-relevant estimates for ion spectra and doses.

Aim 2. Characterize and build computational models for exposure dose and effect modifiers of performance in three behavioral domains, using NASA-provided data sets.

Aim 2a. Evaluate the dose-response and effect-modifier data sets for data quality across ions, exposure regimens, study design, and behavioral outcomes, and for suitability for multivariate modeling of risks for specific behavioral domains. Effect modifiers to be assessed may include strain/genotype, sex, and diet group, depending on the data sets provided.

Aim 2b. Build single-ion risk models that incorporate dose-response and effect-modifier effects for each behavioral domain, using data sets that pass the Aim 2a evaluation criteria.

Aim 2c. Build multi-ion risk models that incorporate dose-response and effect-modifier effects for each behavioral domain, based on the findings of the single ion models of Aim 2b. The risk predictions of the different behavioral domains will be evaluated using both experimental exposure regimens and GCR-relevant estimates for ion spectra and doses.

Aim 3. Apply advanced statistical methods to integrate the risk estimates from Aim 1 and Aim 2 into a unified model to predict individual-level risk for specific behavioral domains.

Aim 4. Evaluate NASA-provided data sets for building computational models for associations of

CNS tissue biomarkers with performance in three behavioral domains.

Aim 4a. Evaluate the CNS tissue biomarker and behavioral outcomes data set for data quality across exposure regimens and behavioral outcomes, and for suitability for multivariate modeling of risks for specific behavioral domains. Effect modifiers that maybe available include strain/genotype and age at exposure. The data sets expected to be available for aim 4 are likely to be single ion experiments with molecular biomarker data from a few CNS sub-regions in animals tested for behavioral outcomes.

Aim 4b. Build single-ion risk models for CNS tissue biomarkers for each behavioral domain, using data sets that pass the Aim 4a evaluation criteria

GSAM project provides delivery scheme of low and high LET particles to best simulate 600/900 day Mars missions within facility and animal constraints.

• Studies required to inform countermeasure testing and validation
• Fluence based versus acute exposures to simulate space radiation 

GSAM (GCR Simulation and Mitigation) research objectives:

Aim 1:  Design and implement Hybrid Shield Concept to simulate GCR environment at NSRL to include production of secondaries and correlated multiplicities found in space

Aim 2:  Validate ground simulation of dose rate including fractionation and time course delivery of particles to simulate low dose chronic exposures in space.  

Aim 3: Evaluation of dose delivery scheme with biological timescales to inform chronic studies.

Aim 4: Expand design and implement Mars surface simulation at NSRL to include mixed field environment found on Mars surface.

• Aim 1 (CVD): Validate surrogate endpoint biomarkers, including standard of care clinical markers, for monitoring space radiation-induced late cardiovascular diseases for use in risk assessment model development and evaluation of protective countermeasures.

• Aim 2 (Late CNS): Define adverse outcome pathways and potential biomarkers associated with CNS functional changes and late neurodegenerative diseases

• Aim 3 (CVD): Evaluate standard of care clinical protective measures, such as exercise, dyslipidemia drugs, anti-inflammatory drugs, diet, as mitigation strategy for late cardiovascular diseases.

• Aim 4 (late CNS): Determine whether CVD countermeasures are effective in mitigating adverse CNS effects.

• Aim 1 (CVD):  Validate efficacy of standard of care clinical protective measures in a combined stressor model with chronic mixed field exposures, evaluating for mitigation of radiation-induced CVD.

• Aim 2 (CVD/CNS): Validate efficacy of cross-risk candidate radioprotective agents in a combined stressor model with chronic mixed field exposures, evaluating for mitigation of radiation-induced CVD and late CNS adverse outcomes.

• Aim 3 (CVD, CNS):  Evaluate any potential antagonistic, synergistic, or additive effects across CVD protective measures and late CNS adverse outcomes.

Specific Aims:

• Aim 1 (Cancer): Determine impact of a simulated 600 day Mars mission exposure on radiation carcinogenesis including tumor burden, spectrum, latency, progression, and aggressiveness.

•  

• Aim 2 (Cancer): Determine impact of a simulated 900 day Mars mission exposure, including complex secondary environment found on Mars Surface, on radiation carcinogenesis including tumor burden, spectrum, latency, progression, and aggressiveness.

•  

• Aim 3 (CVD/CNS): Determine impact of a simulated 600 day Mars mission exposure on cardiovascular and late neurodegenerative CNS diseases.

•  

• Aim 4 (CVD/CNS): Determine impact of a simulated 900 day Mars mission exposure, including complex secondary environment found on Mars Surface cardiovascular and late neurodegenerative CNS diseases.

•  

• Aim 5 (Cross Risk): Validate biomarker approaches for in mission surveillance during Mars 600 and 900 mission simulation.

•  

Studies will focus on doses and dose rates seen on a 600 day free space transit simulating Mars flyby or opposition-class, short stay mission as well as a 900 day conjunction-class, long stay mission on the surface of Mars.

• Aim 1: Determine potential synergistic effects from other spaceflight stressors e.g., psychological (isolation and confinement), altered gravity (micro-gravity), stress, sleep deficiency, altered circadian rhythms, hypercapnea, altered immune, endocrine and metabolic function, or other on CVD and CNS endpoints.

• Aim 2: Quantify impact of combined spaceflight stressors on the spectrum and dose responses for late functional CNS changes related to behavioral changes or degenerative neurological disease associated with space relevant exposures.

•  

Aim 3: Quantify the impact of combined spaceflight stressors on spectrum and dose responses for late functional CVD changes related to clinically relevant endpoints associated with space relevant exposures.
 

Research will evaluate medical countermeasures that mitigate risk of radiation induced diseases addressing multiple tumor types and multiple risk areas (i.e., cancer, late neurodegeneration, cardiovascular disease) including the:

• Mitigation of solid and hematologic cancers due to space radiation exposure as measured by incidence, aggressiveness, overall burden, and time to occurrence.

• Mitigation of cardiovascular disease incidence, latency, and progression as measured by clinical outcomes, events, pathology and survival times.

   

• Mitigation of late neurodegenerative conditions and microvascular disruption of the CNS as measured by changes in performance, cognitive function, and pathology.

Specific Aims:

• Aim 1 (Cross Risk): Identify and prioritize candidate cross-risk countermeasure agents targeting common pathways of adverse outcomes from space radiation exposure

• Aim 2 (Cross Risk): Using appropriate animal models, test efficacy of selected medical countermeasures for prevention of adverse outcomes across multiple risk areas following simulated GCR and SPE exposures.

• Aim 3 (Cross Risk): Identify potential cross-risk and cross-element antagonistic countermeasure approaches

• Aim 4 (Cross Risk): Identify biomarker approaches to assess countermeasure efficacy for surveillance, treatment, and outcome prediction.

Studies to validate identified medical countermeasures that address multiple tumor types and multiple risk areas (i.e., cancer, late neurodegeneration, cardiovascular disease) with the following aims:

Specific Aims:

• Aim 1 (Cross Risk): Validate efficacy of candidate countermeasure agents in additional model systems in compliance with FDA animal rules.

• Aim 2 (Cross Risk): Validate efficacy of candidate countermeasure agents in space relevant environment.

• Aim 3 (Cross Risk):  Determine minimum dosing (mg/day) requirements for optimum effectiveness.

• Aim 4 (Cross Risk):  Evaluate any potential antagonistic effects across space radiation risk area endpoints including multiple tumor types

• Aim 5 (Cross Risk): Evaluate biomarker approaches to assess countermeasure efficacy for surveillance, treatment, and outcome prediction.

Dose-rate and mixed field animal and cellular studies, supporting the development and concept of operations of the NSRL GCR Simulator, are sought to:

Specific Aims:

• Aim 1: Provide data to define the dose delivery scheme for low-LET radiation, the dose delivery scheme for high-LET radiation, and the time course (duration) of cellular and animal studies for simulation of GCR-induced endpoints; including studies to understand the principles governing biological responses to a mixed field of low- and high-LET radiation.

• Aim 2: Reduce uncertainties in dose-rate effects used to estimate risks from space radiation exposures.

Secondary aim:

• Aim 3: Generate mixed field RBEs for relevant endpoints.

Evaluation of dose rate effects and tissue specific RBE’s for major at risk tissues using chronic mixed-field radiation with the following aims:

Specific aims

• Aim 1 (Cancer): Research to improve accuracy of dose and dose-rate effectiveness factor (DDREF) estimates for tumorigenesis and evidence on whether risk is linear or additive over mixed high and low LET radiation exposure

• Aim 2 (Cancer): Determine tissue specific mixed field RBE values for tumorigenesis for major at risk tissues

• Aim 3 (CVD): Examine dose thresholds, dose-rate dependence and mixed field effects on CVD disease latency and progression; evaluate RBEs

• Aim 4 (late CNS): Examine dose thresholds, dose-rate dependence and mixed field effects for late behavioral and neurodegenerative CNS diseases; evaluate RBEs

• Aim 5 (CVD): Evaluate effects of chronic space radiation exposures on ‘standard of care’ clinical markers for CVD effects.

Secondary aims:

• Aim 6: Assess the impact of simulated low dose GCR exposures on the health of the blood forming organs.

Studies will generate mixed field dose response curves to evaluate thresholds and RBE’s.

• Specific Aim A: To facilitate the integration of space radiobiology research results into improved models of BR radiation risk projections.
• Specific Aim B: To complete the cancer risk uncertainty analysis factors to be used for near-term mission planning (trade studies), and to update these as new radiobiological data sets become available.
• Specific Aim C: To develop descriptive and predictive models of modifications to biochemical pathways known likely to be causative of cancer risk.
• Specific Aim D: To integrate or develop new data and models of DNA damage, transmissible chromosomal aberration including translocation, insertions, terminal deletion including telomere loss, and other aberrations related to chromosomal instability or cancer initiation, incorporating applications to the simulations in Specific Aim C.
• Specific Aim E: To perform a comprehensive assessment of available acute radiation risk projection models, and to develop simulation software of acute risk for mission design assessments.