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Dynamic nature of epigenetic patterns observed during the Mars 520-d mission simulation

Interplanetary human spaceflight represents a formidable medical challenge but also provides a unique platform for investigating human adaptation to extreme environmental changes. Understanding the long-term effects of isolation has relevance in a range of scenarios and it is well recognized that a better understanding of the relationship between environmental exposure and the epigenome can lead to more effective preventive measures. Here we conduct a longitudinal epigenetic mood state and biochemical profiling of 6 crew members in an experiment simulating a 520-day mission to Mars. Illumina HumanMethylation450 BeadChip was used to obtain DNA methylation profiles. Firstly we found that long-term isolation can induce global DNA methylation remodeling and this change seems to be an active adaptation (rather than a random process or a by-product of the isolation). This study is the first to demonstrate the dynamic relationship between global epigenetic remodeling and isolation-induced mood state and biochemical changes. Secondly by considering the location of methylation sites within the genome and using gene-pathway annotation we were able to identify pathways that were significantly enriched in methylation events and consider their association with specific function and the timeline of the mission. Thirdly via our definition of epi-entropy a measure of entropy adapted to methylation events we observed that the methylation remodeling produced a marked reduction in epi-entropy. Results suggest that DNA methylation change is an indicator of change rather than its by-product i.e. there is a psychology-epigenome-metabolism model of long-term depression; DNA methylation programs the environment signal into the epigenome which is subsequently transformed into the biochemical output and health outcome. Thus longitudinal epigenetic profiling could code the effect of isolation and act as early indicators of latent health outcome. A longitudinal epigenetic mood state and biochemical profiling of 6 crew members in an experiment simulating a 520-day mission to Mars. 36 samples of blood cell DNA methylation profiling were obtained by Illumina HumanMethylation450 BeadChip across 6 sampling points during the 520 days mission for all of the 6 crew members.

MARS EXPRESS MARS MRS 1/2/3 EXTENDED MISSION 3 3310 V1.0

This is a Mars Express Radio Science data set, collected during the extended mission phase 2010-01-01 to 2012-12-31. It is a Global Gravity measurement and covers the time 2012-07-29T10:05:18.000 to 2012-07-29T13:14:54.000.

MARS EXPRESS MARS MRS 1/2/3 EXTENDED MISSION 5 3806 V1.0

This is a Mars Express Radio Science data set, collected during the extended mission phase 2015-01-01 to 2016-12-31. It is a Occultation measurement and covers the time 2015-09-01T14:10:09 to 2015-09-02T06:48:38.999.

Transcriptional profiling of spleens from mice flown on the RR-9 mission

The objective of the Rodent Research-9 (RR-9) mission was to use mice to understand the molecular basis of phenomena that affect astronauts during long-duration spaceflight, particularly visual impairment, and joint tissue degradation. To this end, a flight group (FLT) of 10-week-old male C57BL/6J mice were launched from Kennedy Space Center (KSC) on 8/14/2017 and housed in Rodent Habitats on the ISS for 33 days before being returned alive to Earth. After splashdown in the Pacific Ocean, the animals were transported to Loma Linda University (LLU) for testing, euthanasia, and dissection on 9/18/2018. Ground Control (GC) studies were planned to commence at KSC approximately one-week after the conclusion of the flight experiments. However, all the GC mouse studies at KSC had to be cancelled due to Hurricane Irma and potential adverse effects on the animal housing facility. The GC studies were therefore rescheduled and begun in May 2018. The GC was euthanized and dissected 6/18/2018 - 6/20/2018. Because this resulted in a different cohort of mice being used for the GC controls as compared to the flight (FLT) groups, two cohort controls were included in the study. The first, Cohort Control 1 (CC_C1), was from the same cohort as the FLT animals and was sacrificed and dissected 4 days after the FLT group (9/22/2017). The second, Cohort Control 2 (CC_C2), was from the same cohort as the GC, and was sacrificed and dissected 2-8 days after the GC, (6/24/2018 - 6/26/2018). The CC_C1 and CC_C2 groups were housed in standard cages and fed standard chow in contrast to all other groups which received Rodent Foodbars. Upon dissection, spleen tissues were preserved in liquid nitrogen and stored at 80 C before RNA was extracted. Only the flight (FLT, n of 10) and Ground Control (GC, n of 10) samples were processed and analyzed in this study. Libraries were generated using a 3’ Tag-seq approach and sequenced at a targeted depth of 40 M clusters (SE 93 bp).

MARS EXPRESS MARS PFS EDR FOURTH EXTENSION MISSION DATA V1.0

The Mars Express PFS data set contains raw (CODMAC Level 2) measurements from the Planetary Fourier Spectrometer collected during the fourth extension Mars orbit phases.

Bulk RNA sequencing and spatially resolved transcriptional profiling of cerebellums from mice flown on the RRRM-2 mission

In the Rodent Research Reference Mission (RRRM-2), forty female C57BL/6NTac mice were flown on the International Space Station. To assess differences in outcomes due to age, twenty 12 week-old and twenty 29 week-old mice were flown, respectively. To directly assess spaceflight effects, half of the young and old mice (10 old, 10 young) were sacrificed on-orbit after 55-58 days (ISS Terminal, ISS-T), while the other half (10 old, 10 young) were returned live to Earth after 32 days and allowed to recover for 24 days (Live Animal Return, LAR) before sacrifice. ISS-T and LAR mice were the same age at sacrifice. Both the ISS-T and LAR animals had independent ground controls (10 mice per group housed in flight hardware in matched environmental conditions), basal controls (10 mice per group sacrificed 2 days before launch), and vivarium controls (10 mice per group housed within standard vivarium habitats). Thus RRRM-2 included a total of 160 mice. This study includes bulk RNA sequencing and spatially resolved transcriptional profiling data from cerebellums from 4 (bulk RNAseq) or 5 (spatial transcriptomics) old ISS-T flight animals, 3 old ISS-T ground control (GC) animals, 5 young ISS-T flight animals, 3 young ISS-T GC animals, 3 old LAR flight animals, 3 old LAR GC animals, 2 (bulk RNAseq) or 3 (spatial transcriptomics) young LAR flight animals, and 3 young LAR GC animals. Cerebellums from the right hemisphere were embedded and cryosectioned. Cryosections were either processed for bulk RNA sequencing or placed on gene expression arrays, stained and imaged. Imaging was followed by tissue permeabilization to release mRNA molecules from cells for capture onto the array surface. Subsequently, spatial transcriptomics libraries were prepared and sequenced.

MARS EXPRESS MARS MRS 1/2/3 EXTENDED MISSION 3 3339 V1.0

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MARS EXPRESS MARS MRS 1/2/3 EXTENDED MISSION 5 3816 V1.0

Transcriptional profiling of cerebellums from mice flown on the RR-23 mission

The objective of the Rodent Research-23 missions (RR-23) was to better understand the effects of spaceflight on the eyes, specifically on the structure and function of the arteries, veins, and lymphatic vessels that are needed to maintain vision. To this end, twenty male, C57BL/6J, 16-17 weeks old mice were delivered to the ISS on SpaceX-21 in a single transporter, transferred to two rodent habitats, and maintained in microgravity for 38 days. Flight mice were then returned to Earth alive (Jan 13th, 2021). After splashdown in the Atlantic Ocean, mice were transported to Kennedy Space Center via helicopter. The 20 Flight, 20 Habitat Ground Control (HGC), and 20 Vivarium Ground Control (VGC) mice were removed from Rodent Transporters (Flight and HGC) or vivarium cages (VGC), placed into shipping containers, and flown to Texas A and M University. There, mice underwent post flight procedures, before euthanasia and tissue collection. Flight, HGC and VGC animals were euthanized and dissected on Jan 14th, 17th or 20th of 2021, respectively. Cerebellums were preserved by immersion in RNAlater and stored at -80 ˚C until RNA was extracted, and libraries generated and sequenced (target 60 M clusters per sample, PE 150 bp). This dataset features 9 samples from the Flight group, 9 samples from the HGC group, and 8 samples from the VGC group. A technical replicate is included for one sample in each group. These consist of an independent library preparation for a single RNA extraction.

GIOTTO EXTENDED MISSION ELECTRON PARTICLE ANALYSER V1.0

This dataset contains count rates from the Energetic Particle Analyser (EPA) experiment aboard the Giotto spacecraft during its flyby of comet P/Grigg-Skjellerup during July of 1992. These data were not formally reviewed by the PDS.

Transcriptional profiling of livers from mice flown on the RR-23 mission

The objective of the RR-23 mission was to better understand the effects of spaceflight on the eyes, specifically on the structure and function of the arteries, veins, and lymphatic vessels that are needed to maintain vision. To this end, twenty male, C57BL/6J, 16-17 weeks-old mice were delivered to the ISS on SpaceX-21 in a single transporter, transferred to two rodent habitats, and maintained in microgravity for 38 days. Flight mice were then returned to Earth alive (Jan 13th, 2021). After splashdown in the Atlantic Ocean, mice were transported to Kennedy Space Center via helicopter. The 20 Flight, 20 Habitat Ground Control (HGC), and 20 Vivarium Ground Control (VGC) mice were removed from Rodent Transporters (Flight and HGC) or vivarium cages (VGC), placed into shipping containers, and flown to Texas A and M University. There, mice underwent post-flight procedures, before euthanasia and tissue collection. Flight, HGC and VGC animals were euthanized and dissected on Jan 14th, 17th or 20th of 2021, respectively. Livers were preserved by flash freezing in liquid nitrogen and stored at -80 C until RNA was extracted, and libraries generated and sequenced (target 60 M clusters per sample, PE 150 bp). This dataset features 9 samples from the Flight and VGC groups, and 8 samples from the HGC group.

Transcriptional analysis of kidneys from mice flown on the RR-7 mission

The objective of the Rodent Research-7 mission (RR-7) was to study the impact of the space environment on the gut microbiota of two strains of mice and how any changes in-turn affect the immune system, metabolic system, and circadian or daily rhythms. To this end, ten 11-week-old female C57BL/6J and ten 11-week-old female C3H/HeJ mice were flown to the International Space Station on June 29, 2018 on SpaceX-15 and housed in two Rodent Habitats. Samples of food, swabs from living surfaces, and fecal pellets were collected from each animal before launch and regularly during the mission. The mission also involved extended video collection (48 hr video segments per Habitat) to monitor circadian rhythms, and on-orbit mass measurement. After 25 days on-orbit, half of the mice of each strain were euthanized on the ISS with Ketamine/Xylazine/Acepromazine and cardiac puncture, after which carcasses were segmented in three sections and preserved in RNA later. After 75-76 days the remaining 5 animals from each group were euthanized and processed in the same manner. The 25-day dissected carcasses returned on SpX-15, and the 75-day dissected carcasses returned on SpX-16. In addition to the Flight group, three ground control groups were also part of the study: Basal (representing the pre-launch state), Vivarium (standard vivarium housing for the same duration of time as flight), and Ground (same habitat in the International Space Station Environment Simulator, ISSES). Twenty mice (10 of each strain) were included in each of these control groups, which were euthanized and processed on the same schedule and in the same manner as the flight samples. Dissections for tissues from all experimental groups were completed by the PI groups along with NASA's Biospecimen Sharing Program in February 2019. GeneLab received kidneys from thirty-eight C57BL/6J mice: 9 Basal, 5 Ground (25 days), 4 Ground (75 days), 5 Flight (25 days), 5 Flight (75 days), 5 Vivarium (25 days), 5 Vivarium (75 days). GeneLab received kidneys from thirty-nine C3H/HeJ mice: 9 Basal, 5 Ground (25 days), 5 Ground (75 days), 5 Flight (25 days), 5 Flight (75 days), 5 Vivarium (25 days), 5 Vivarium (75 days). From these kidneys, RNA was extracted, libraries generated (stranded, ribodepleted) and sequenced (target 60 M clusters at PE 98 bp).

MARS EXPRESS MARS MRS 1/2/3 EXTENDED MISSION 3 3018 V1.0

This is a Mars Express Radio Science data set, collected during the extended mission phase 2010-01-01 to 2012-12-31. It is a Global Gravity measurement and covers the time 2011-07-19T00:02:46.000 to 2011-07-19T00:43:29.500.

MARS EXPRESS MARS MRS 1/2/3 EXTENDED MISSION 3 3397 V1.0

MARS EXPRESS MARS MRS 1/2/3 EXTENDED MISSION 5 3718 V1.0

This is a Mars Express Radio Science data set, collected during the extended mission phase 2015-01-01 to 2016-12-31. It is a Occultation measurement and covers the time 2015-04-04T00:07:34 to 2015-04-05T13:31:42.500.