NOTE: End date changed to 08/31/2023 per TRISH (Ed., 2/23/23)

Astronauts will soon be deploying on deep space missions to Mars, and will be exposed to continuous low dose, high energy ionizing radiation (IR). This IR is different from Earth orbit radiation; therefore, this kind of space radiation may lead to undefined acute and chronic health problems from continuous radiation exposure. Previous research on the effects of space radiation have predominantly studied male astronauts. Data from cancer patients receiving radiation, or animal research, suggest women might show an exaggerated inflammation response, making them more likely to develop diseases of the heart or blood vessels (vascular cells). These unknown sex differences remain as unexplored as deep space. The purpose of this fellowship proposal is to investigate how men and women differ in their immune and vascular response to IR, as radiation may increase or accelerate disease burden in astronauts who traveled into deep space. The first study goal is to test increasing levels of IR to see if the inflammation response is more sensitive in one sex versus the other. Immune cells from healthy adults will be exposed to IR and followed for 24 hours. Measurements include signals the cells produce, signals inside and on the surface of immune cells that change their function. In the second study, immune and vascular cells are placed together after IR, to see if crosstalk between cells is different between the sexes. Methods from study one will be repeated in both cell types, plus measuring adhesion molecules on the surface of vascular cells that allow immune cells to bind and amplify inflammation. Crosstalk is expected to magnify the inflammation response. The findings are expected to help develop sex-specific interventions to minimize risk from IR, so astronaut health and mission success are maximized.

Aim 2 was to determine if the immune cells response to stimulus is impacted post acute ionizing proton radiation. We showed that 48 hours after 1Gy of radiation the expression of anti-/pro-inflammatory cytokines is increased in a similar manner between sexes in response to a stimulus. We also found that 72 hours after 1 Gy of radiation there was no sex difference in the response of anti-/pro-inflammatory cytokines. There was also a similar increase in stimulated cytokine expression after 72 hours 1Gy compared to 48 hours. However, we found that 72 hours post 1 Gy of radiation had a greater stimulated cytokine expression compared to 48 hours.

Future studies include increasing our sample size, utilizing different dosages of proton radiation, performing repeated dosages of proton radiation, and using flow-cytometry to determine the intracellular immune and sex dependent response of macrophages.

This project showed that immune cells that are exposed to proton radiation, which is the predominate form of space radiation, produce inflammatory markers in a sex specific manner. We also showed that the immune cells may have an exaggerated response to a stimulus after they have irradiated with proton radiation.

This information indicates that the immune cells exposed to an acute dose radiation have a lower stimulus threshold. This can lead to an exaggerated immune response to a viral or bacterial stimulus. Countermeasures to the priming of the immune cells could be deployed or developed to prevent the immune cells from being overstimulated.

NOTE: End date changed to 08/31/2023 per TRISH (Ed., 2/23/23)

Astronauts will soon be deploying on deep space missions to Mars, and will be exposed to continuous low dose, high energy ionizing radiation (IR). This IR is different from Earth orbit radiation; therefore, this kind of space radiation may lead to undefined acute and chronic health problems from continuous radiation exposure. Previous research on the effects of space radiation have predominantly studied male astronauts. Data from cancer patients receiving radiation, or animal research, suggest women might show an exaggerated inflammation response, making them more likely to develop diseases of the heart or blood vessels (vascular cells). These unknown sex differences remain as unexplored as deep space. The purpose of this fellowship proposal is to investigate how men and women differ in their immune and vascular response to IR, as radiation may increase or accelerate disease burden in astronauts who traveled into deep space. The first study goal is to test increasing levels of IR to see if the inflammation response is more sensitive in one sex versus the other. Immune cells from healthy adults will be exposed to IR and followed for 24 hours. Measurements include signals the cells produce, signals inside and on the surface of immune cells that change their function. In the second study, immune and vascular cells are placed together after IR, to see if crosstalk between cells is different between the sexes. Methods from study one will be repeated in both cell types, plus measuring adhesion molecules on the surface of vascular cells that allow immune cells to bind and amplify inflammation. Crosstalk is expected to magnify the inflammation response. The findings are expected to help develop sex-specific interventions to minimize risk from IR, so astronaut health and mission success are maximized.

The only remaining steps from year 1 to complete are actual radiation exposure of cells (1 week) with follow-up immunology assays and analysis (2 weeks); thus, we feel confident we are largely on track for this fellowship. We have increased recruitment for the biospecimen samples during the second half of the year and are currently at 100% of male and 73% of female samples collected.

Looking forward toward Year 2, we aim to:

• Submit at least 2 manuscripts from Aim 1 data.

• Learn laboratory culture practices to grow endothelial cells and study the interactions between endothelial cells and immune cells-the goal for Aim 2.

• Complete the second round of irradiation (Year 2) allowing flexibility for follow-up or confirmation experiments based on Aim 1 findings.

• Repeat radiation studies on other immune cell types for preliminary data for future grants.

Astronauts will soon be deploying on deep space missions to Mars, and will be exposed to continuous low dose, high energy ionizing radiation (IR). This IR is different from Earth orbit radiation; therefore, this kind of space radiation may lead to undefined acute and chronic health problems from continuous radiation exposure. Previous research on the effects of space radiation have predominantly studied male astronauts. Data from cancer patients receiving radiation, or animal research, suggest women might show an exaggerated inflammation response, making them more likely to develop diseases of the heart or blood vessels (vascular cells). These unknown sex differences remain as unexplored as deep space. The purpose of this fellowship proposal is to investigate how men and women differ in their immune and vascular response to IR, as radiation may increase or accelerate disease burden in astronauts who traveled into deep space. The first study goal is to test increasing levels of IR to see if the inflammation response is more sensitive in one sex versus the other. Immune cells from healthy adults will be exposed to IR and followed for 24 hours. Measurements include signals the cells produce, signals inside and on the surface of immune cells that change their function. In the second study, immune and vascular cells are placed together after IR, to see if crosstalk between cells is different between the sexes. Methods from study one will be repeated in both cell types, plus measuring adhesion molecules on the surface of vascular cells that allow immune cells to bind and amplify inflammation. Crosstalk is expected to magnify the inflammation response. The findings are expected to help develop sex-specific interventions to minimize risk from IR, so astronaut health and mission success are maximized.