Spaceflight induces molecular, cellular and physiological shifts in astronauts and poses myriad biomedical challenges to the human body, which are becoming increasingly relevant as more humans venture into space(1-6). Yet current frameworks for aerospace medicine are nascent and lag far behind advancements in precision medicine on Earth, underscoring the need for rapid development of space medicine databases, tools and protocols. Here we present the Space Omics and Medical Atlas (SOMA), an integrated data and sample repository for clinical, cellular and multi-omic research profiles from a diverse range of missions, including the NASA Twins Study(7), JAXA CFE study(8,9), SpaceX Inspiration4 crew(10-12), Axiom and Polaris. The SOMA resource represents a more than tenfold increase in publicly available human space omics data, with matched samples available from the Cornell Aerospace Medicine Biobank. The Atlas includes extensive molecular and physiological profiles encompassing genomics, epigenomics, transcriptomics, proteomics, metabolomics and microbiome datasets, which reveal some consistent features across missions, including cytokine shifts, telomere elongation and gene expression changes, as well as mission-specific molecular responses and links to orthologous, tissue-specific mouse datasets. Leveraging the datasets, tools and resources in SOMA can help to accelerate precision aerospace medicine, bringing needed health monitoring, risk mitigation and countermeasure data for upcoming lunar, Mars and exploration-class missions.

Background:The Inspiration4 (I4) mission, the first all-civilian orbital flight mission, investigated the physiological effects of short-duration spaceflight through a multi-omic approach. Despite advances, there remains much to learn about human adaptation to spaceflight's unique challenges, including microgravity, immune system perturbations, and radiation exposure. Methods:To provide a detailed genetics analysis of the mission, we collected dried blood spots pre-, during, and post-flight for DNA extraction. Telomere length was measured by quantitative PCR, while whole genome and cfDNA sequencing provided insight into genomic stability and immune adaptations. A robust bioinformatic pipeline was used for data analysis, including variant calling to assess mutational burden. Result:Telomere elongation occurred during spaceflight and shortened after return to Earth. Cell-free DNA analysis revealed increased immune cell signatures post-flight. No significant clonal hematopoiesis of indeterminate potential (CHIP) or whole-genome instability was observed. The long-term gene expression changes across immune cells suggested cellular adaptations to the space environment persisting months post-flight. Conclusion:Our findings provide valuable insights into the physiological consequences of short-duration spaceflight, with telomere dynamics and immune cell gene expression adapting to spaceflight and persisting after return to Earth. CHIP sequencing data will serve as a reference point for studying the early development of CHIP in astronauts, an understudied phenomenon as previous studies have focused on career astronauts. This study will serve as a reference point for future commercial and non-commercial spaceflight, low Earth orbit (LEO) missions, and deep-space exploration.

The SpaceX Inspiration4 mission provided a unique opportunity to study the impact of spaceflight on the human body. Biospecimen samples were collected from the crew at different stages of the mission, including before (L-92, L-44, L-3 days), during (FD1, FD2, FD3), and after (R+1, R+45, R+82, R+194 days) spaceflight, creating a longitudinal sample set. The collection process included samples such as venous blood, capillary dried blood spot cards, saliva, urine, stool, body swabs, capsule swabs, SpaceX Dragon capsule HEPA filter, and skin biopsies, which were processed to obtain aliquots of serum, plasma, extracellular vesicles, and peripheral blood mononuclear cells. All samples were then processed in clinical and research laboratories for optimal isolation and testing of DNA, RNA, proteins, metabolites, and other biomolecules. This paper describes the complete set of collected biospecimens, their processing steps, and long-term biobanking methods, which enable future molecular assays and testing. As such, this study details a robust framework for obtaining and preserving high-quality human, microbial, and environmental samples for aerospace medicine in the Space Omics and Medical Atlas (SOMA) initiative, which can also aid future experiments in human spaceflight and space biology.

Spaceflight induces an immune response in astronauts. To better characterize this effect, we generated single-cell, multi-ome, cell-free RNA (cfRNA), biochemical, and hematology data for the SpaceX Inspiration4 (I4) mission crew. We found that 18 cytokines/chemokines related to inflammation, aging, and muscle homeostasis changed after spaceflight. In I4 single-cell multi-omics data, we identified a "spaceflight signature" of gene expression characterized by enrichment in oxidative phosphorylation, UV response, immune function, and TCF21 pathways. We confirmed the presence of this signature in independent datasets, including the NASA Twins Study, the I4 skin spatial transcriptomics, and 817 NASA GeneLab mouse transcriptomes. Finally, we observed that (1) T cells showed an up-regulation of FOXP3, (2) MHC class I genes exhibited long-term suppression, and (3) infection-related immune pathways were associated with microbiome shifts. In summary, this study reveals conserved and distinct immune disruptions occurring and details a roadmap for potential countermeasures to preserve astronaut health. Multiple omics platforms and deep single-cell profiling in the I4 astronauts reveal both conserved and distinct immune system disruptions across missions, provide a single-cell immune reference for future missions.

Breast cancer imposes a significant burden globally. While the survival rate is steadily improving, much remains to be elucidated. This observational, single time point, multiomic study utilizing genomics, proteomics, targeted and untargeted metabolomics, and metagenomics in a breast cancer survivor (BCS) and age-matched healthy control cohort (N = 100) provides deep molecular phenotyping of breast cancer survivors. In this study, the BCS cohort had significantly higher polygenic risk scores for breast cancer than the control group. Carnitine and hexanoyl carnitine were significantly different. Several bile acid and fatty acid metabolites were significantly dissimilar, most notably the Omega-3 Index (O3I) (significantly lower in BCS). Proteomic and metagenomic analyses identified group and pathway differences, which warrant further investigation. The database built from this study contributes a wealth of data on breast cancer survivorship where there has been a paucity, affording the ability to identify patterns and novel insights that can drive new hypotheses and inform future research. Expansion of this database in the treatment-naïve, newly diagnosed, controlling for treatment confounders, and through the disease progression, can be leveraged to profile and contextualize breast cancer and breast cancer survivorship, potentially leading to the development of new strategies to combat this disease and improve the quality of life for its victims.

Ten years ago, it was predicted that the multi-omics revolution would also revolutionize space pharmacogenomics. Current barriers related to the findable, accessible, interoperable, and reproducible use of space-flown pharmaceutical data have contributed to a lack of progress beyond application of earth-based principles. To directly tackle these challenges, we have produced a novel database of all the drugs flown into space, compiled from publicly available ontological and spaceflight-related datasets, to exemplify analyses for describing significant spaceflight-related targets. By focusing on mechanisms perturbed by spaceflight, we have provided a novel avenue for identifying the most relevant changes within the drug absorption, distribution, metabolism, and excretion pathways. We suggest a set of space genes, by necessity limited to available tissue types, that can be expanded and modified based on future tissue-specific and mechanistic-specific high-throughput assays. In sum, we provide the justification and a definitive starting point for pharmacogenomics guided spaceflight as a foundation of precision medicine, which will enable long-term human habitation of the Moon, Mars, and beyond. ![Figure][1] ### Competing Interest Statement CEM is a co-Founder of Onegevity, Twin Orbit, and Cosmica Biosciences. CMS, JCS, and MAS are owners in Sovaris Holdings, LLC. [1]: pending:yes

Background: Military special operators, elite athletes, and others requiring uninterrupted optimal performance currently lack options for sleep and mood support without performance-inhibiting effects. Kavalactones, derived from the root of the kava plant (Piper methysticum Forst), have been shown to elevate mood and wellbeing by producing a feeling of relaxation without addiction or cognitive impairment. Methods: In this placebo-controlled, crossover study (NCT05381025), we investigated the effects of 2 weeks of kavalactones use on cortisol (diurnal salivary), sleep (RSQ-W; Restorative Sleep Questionnaire, Weekly), mood (DASS-21; Depression Anxiety Stress Scale-21), and motivation state to expend (Move) or conserve (Rest) energy (CRAVE; Cravings for Rest and Volitional Energy Expenditure, Right Now) in a cohort of 15 healthy, physically fit young males engaged in a rigorous, two-a-day preparation class for special operations forces qualification. Results: Cortisol, sleep, and mood were within normal, healthy parameters in this cohort at baseline. This remained unchanged with kavalactones use with no significant findings of clinical interest. However, a statistically similar, positive slope for within-group Move scores was seen in both groups during kavalactones loading (first group Move slope 2.25, second group Move slope 3.29, p = 0.299). This trend was seen regardless of order and with no apparent effects on the Rest metric (all p >= 0.05). Moreover, a significant between-group difference appeared after 1 week of kavalactones use in the first phase (p = 0.044) and persisted through the end of the first loading period (p = 0.022). Following the 10-day washout, this between-groups divergence remained significant (p = 0.038) but was reversed by 1 week after the crossover (p = 0.072), with Move scores once again statistically similar between groups and compared to baseline at study end. Furthermore, the group taking kavalactones first never experienced a significant decrease in Move motivation state (lowest mean score 21.0, highest 28.6, all p >= 0.05), while the group receiving kavalactones in the last 2 weeks of the study had Move scores that were statistically lower than baseline (lowest mean score 8.6, highest 25.9, all p <= 0.05) at all time points but the last (p = 0.063) after 2 weeks of kavalactones exposure. Conclusions: We report a novel finding that kavalactones may support performance by maintaining or rescuing the desire to expend energy in the context of significant physical and mental strain in well-conditioned individuals, even in a context of already normal cortisol, sleep, and mood.

Abstract The I4 mission, the first all-civilian orbital flight mission, investigated the physiological effects of short duration spaceflight through a multi-omic approach. We analyzed telomere length, clonal hematopoiesis of indeterminate potential (CHIP), whole genome stability, cell-free DNA (cfDNA) cell lysis, and immune cell gene expression. Our results revealed telomere length dynamics similar to those observed in the NASA Twins Study and in astronauts spending 6 months on the ISS, with lengthening in space and shortening upon return to Earth. Our cell-type of origin analysis of cfDNA fragments revealed an increased presence of innate and adaptive immune cell signatures that persisted over a month after return to earth. No significant relationship between spaceflight and CHIP-related or whole genome abnormalities were observed. Longitudinal mitochondrial, ribosomal and immune function gene expression changes occurred across both adaptive and innate immune cells, suggesting adaptations to the space environment can extend months after return from spaceflight and alter immune function. Our findings provide valuable insights into the physiological consequences of short duration spaceflight and will serve as a reference point for future space tourism, low Earth-orbit (LEO) missions, and deep-space exploration.

PURPOSE: This pilot crossover study investigated the effect of kavalactones (KL) on selected biological and behavioral measures that impact performance and recovery associated with heavy training, including motivation to expend energy. METHODS: 15 healthy, physically fit males (mean age 22.7y) in a rigorous Special Forces preparation class took 225 mg KL then placebo (K-P) or placebo then KL (P-K) for 14 days each with 10 days intervening washout. Motivation to expend energy (Move) was monitored weekly during loading with the validated CRAVE scale (Cravings for Rest and Volitional Energy Expenditure; 0-50 range, higher is more motivated). RESULTS: A significant difference in Move was observed between K-P and P-K (between group) after the first week (L1m: 14.4 vs 26.5, p = 0.026) and remained so through three time points (L2s: 8.6 vs 21.0, p = 0.018). A significant within-group decrease in Move was seen in P-K while taking placebo at study start (25.9 → 14.4, p = 0.047), while within group changes observed in K-P did not reach significance (24.1 → 26.5, p = 0.726). (Figure 1) CONCLUSIONS: We report a novel finding that 225 mg KL at night for two weeks appears to promote and restore the desire to expend energy in the context of substantial physical and mental strain in trained males. KL used at the start of a period of increased exertion appeared to preserve motivation with possible lasting effect, while participants on placebo lost motivation. Use of KL following loss of motivation appeared to restore motivation within a two-week period. This finding was coupled with similar KL effect trajectories regardless of whether KL was ingested in the first or the second phase of the study, as highlighted within each slope of the Kava treatment arms. We conclude that KL may be useful in supporting motivation in elite athletes, though further investigation is needed.Figure 1.: Mean CRAVE Move Scores. ■ = K-P, ● = P-K. †p ≤ 0.05 (t-test) K-P vs P-K. *p ≤ 0.05 (Wilcoxon) vs P-K at baseline (L1s).

Abstract The I4 mission, the first all-civilian orbital flight mission, investigated the physiological effects of short duration spaceflight through a multi-omic approach. We analyzed telomere length, clonal hematopoiesis of indeterminate potential (CHIP), whole genome stability, cell-free DNA (cfDNA) cell lysis, and immune cell gene expression. Our results revealed telomere length dynamics similar to those observed in the NASA Twins Study and in astronauts spending 6 months on the ISS, with lengthening in space and shortening upon return to Earth. Our cell-type of origin analysis of cfDNA fragments revealed an increased presence of innate and adaptive immune cell signatures that persisted over a month after return to earth. No significant relationship between spaceflight and CHIP-related or whole genome abnormalities were observed. Longitudinal mitochondrial, ribosomal and immune function gene expression changes occurred across both adaptive and innate immune cells, suggesting adaptations to the space environment can extend months after return from spaceflight and alter immune function. Our findings provide valuable insights into the physiological consequences of short duration spaceflight and will serve as a reference point for future space tourism, low Earth-orbit (LEO) missions, and deep-space exploration.

BackgroundFactors influencing individual susceptibility to brain acceleration forces or to poor outcomes in brain injury are not well understood. Characterization of blood variants in athletes entering the highly competitive contact environment of professional football, when coupled with longitudinal follow-up of future concussion incidence and trajectory, may provide additional insight into factors that influence brain injury. We observed the metabolic phenotype of collegiate football players entering the 2016 National Football League (NFL) draft. The principal aims were to characterize the molecular status of individual athletes and quantify the prevalence of athletes with multiple concurrent molecular deficits. MethodsBlood was taken from 30 elite American collegiate football players seven weeks before the NFL scouting combine and 15-weeks before entering the NFL draft. ResultsAverage results revealed undesirable values in Omega-3 Index (avg ± std, 4.66 ± 1.16 %), AA:EPA fatty acid ratio (29.13 ± 10.78), homocysteine (11.4 ± 3.4 µmol/L), vitamin D (30 ± 11.4 ng/mL), and RBC magnesium (4.1 ± 0.8 mg/dL). Using sport optimized reference ranges from previously published research, 10% presented with 3, 40% presented with 4, and 50% of athletes presented with 5 undesirable values at once. ConclusionWe conclude molecular deficits in this cohort entering the NFL draft were common, with a significant number of athletes presenting with multiple abnormalities, all with relevant influence on brain health and function. This data warrants extensive metabolic phenotyping and prophylactic precision nutrition countermeasures for athletes entering contact environments.

In this Commentary, we will discuss some of the current trends and challenges in modeling microbiome metabolism. A focus will be the state of the art in the integration of metabolic networks, ecological and evolutionary principles, and spatiotemporal considerations, followed by envisioning integrated frameworks incorporating different principles and data to generate predictive models in the future.

Metabolites produced by enteric microbes may have important effects on astronauts on long-duration missions. The NASA Twins Study provided the most comprehensive multi-scale omics data to date from which to extract molecular features of potential clinical significance to spaceflight. From the multivariate data, we identified an elevation of the uremic toxin p -cresol, which is produced by gut microbial fermentation of dietary tyrosine. p -Cresol has adverse metabolic effects via depletion of the hepatic sulfur pool, which impacts metabolism of drugs, endogenous metabolites, and xenobiotics. Moreover, p -cresol reshapes gut microbial community structure by facilitating survival of species such as clostridia and inhibition of butyrate producers. Spaceflight may also impact the genes responsible for the metabolism of p -cresol (e.g. SULT) and for the safe metabolism of common drugs used in space, such as acetaminophen (e.g. SULT, CYP4502E1, GST). Understanding p -cresol production and its related molecular networks in astronauts may lead to precision medicine advances that enhance astronaut safety and performance on long-duration missions.

Background: At present, there is no clear understanding of the effect of long-duration spaceflight on the major enzymes that govern the metabolism of omega-6 and omega-3 fatty acids. To address this gap in knowledge, we used data from the NASA Twins Study, which includes a multiscale omics investigation of the changes that occurred during a year-long (340 days) human spaceflight. Embedded within the NASA Twins data are specific analytes associated with fatty acid metabolism. Objectives: To examine the long-chain fatty acid desaturases and elongases in a single human during 1 year in space. Method: One male twin was on board the International Space Station (ISS) for 1 year, while his monozygotic twin served as a genetically matched ground control. Longitudinal assessments included the genome, epigenome, transcriptome, proteome, metabolome, microbiome, and immunome during the mission, as well as 6 months before and after. The gene-specific fatty acid desaturase and elongase transcriptome data (FADS1, FADS2, ELOVL2, and ELOVL5) were extracted from untargeted RNA-seq measurements derived from white blood cell fractions. Results: Most data from the elongases and desaturases exhibited relatively similar expression profiles (R-2 > 0.6) over time for the CD8, CD19, and lymphocyte-depleted (LD) cell fractions, indicating overall conservation of function within and between the subjects. Both cell-type and temporal specificity was observed in some cases, and some differences were also apparent between the polyadenylated (polyA) fraction of processed RNAs versus the ribodepleted (ribo-) fraction. The flight subject showed a stronger enrichment of the fatty acid metabolic process pathway across almost all cell types (col-umns, CD4, CD8, CPT, and LD), most especially in the ribodepleted fraction of RNA, but also with the polyA+ fraction of RNA. Gene set enrichment analysis (GSEA) measures across three related fatty acid metabolism pathways showed a differential between the ground and the flight subject. Conclusions: There appears to be no persistent alteration of desaturase and elongase gene expression associated with 1 year in space. However, these data provide evidence that cellular lipid metabolism can be responsive and dynamic to spaceflight, even though it appears cell-type and context specific, most notably in terms of the fraction of RNA measured and the collection protocols. These results also provide new evidence of mid-flight spikes in expression of selected genes, which may indicate transient responses to specific insults during spaceflight. (C) 2020 The Author(s) Published by S. Karger AG, Basel

To date, more than 565 professional astronauts have flown in space, of whom only 11% have been women. Of these, ∼565 have been NASA astronauts, of whom 86% have been men and 14% have been women. Th...

The large C2H2-Zinc Finger (C2H2-ZNF) gene family has rapidly expanded in primates through gene duplication. There is consequently considerable sequence homology between family members at both the nucleotide and amino acid level, allowing for coordinated regulation and shared functions. Here we show that multiple C2H2-ZNF mRNAs experience differential polyadenylation resulting in populations with short and long poly(A) tails. Furthermore, a significant proportion of C2H2-ZNF mRNAs are retained in the nucleus. Intriguingly, both short poly(A) tails and nuclear retention can be specified by the repeated elements that encode zinc finger motifs. These Zinc finger Coding Regions (ZCRs) appear to restrict polyadenylation of nascent RNAs and at the same time impede their export. However, the polyadenylation process is not necessary for nuclear retention of ZNF mRNAs. We propose that inefficient polyadenylation and export may allow C2H2-ZNF mRNAs to moonlight as non-coding RNAs or to be stored for later use.