Biological risks associated with space radiation and microgravity are major concerns for long-term space travel. Through a Systems Biology approach our previous NASA work has shown both TGF xce xb2 signaling pathways and miRNAs have a critical impact on defining health risks with and without space irradiation. We hypothesize that circulating microRNA (miRNA) signatures are driving microvascular disease and muscle degeneration associated with accelerating aging and will be enhanced by exposure to the space environment (radiation and microgravity). We investigated this hypothesis both in vivo and in vitro and test novel antagonist therapies to these miRNA signatures as countermeasures to reduce space radiation-induced health risks. A comprehensive Systems Biology approach was used to examine the influence by high atomic number by high (H) atomic number (Z) and energy (E) (HZE) irradiation. To simulate low-dose exposure due to galactic cosmic rays (GCR) we used the ions energy and doses determined by a NASA consensus formula of 7 different ions to represent GCR (referred to as GCR sim model). To simulate high-dose radiation exposure due to solar particle events (SPE) we used an acute dose of SPE simulated beam at 1Gy which has energies ranging from 50MeV to 150MeV. C57BL/6 wild-type mice were utilized for irradiation with our established simulated microgravity model (hindlimb suspension model) and an in vitro 3D microvasculature tissue model under simulated microgravity (clinostat) conditions will also be irradiated. To expand on the circulating miRNA signature determined from our preliminary data we determined a group of conserved miRNAs which are commonly being regulated in the majority of the organs and tissues throughout the host using our established techniques. MiRNA-sequencing on serum (before IR and at time of sacrifice) liver heart and muscle tissue for all radiation groups revealed the key circulating miRNA signature (consisting of multiple miRNAs) impacting disease risk. Collectively understanding of how whole body space radiation impacts microvascular and tissue degeneration through circulating miRNAs will greatly enhance health risk prognostication and provide possible new mechanisms for protection against space radiation.

The thymus undergoes atrophy during spaceflight. In this study we analyzed gene expression of the thymus of mice on board International space station to elucidate molecular aspects of the thymic atrophy by spaceflight.

This study s goal was to examine if any salivary microbiome changes were observed in astronauts as a result of spaceflight. In addition this study looked for any microbiome signature that may be associated with viral reactivation in humans

Understanding plant adaptive responses to the space environment is a requisite for enabling space farming. Spaceflight produce deleterious effects on plant cells particularly affecting ribosome biogenesis a complex stress-sensitive process coordinated with cell division and differentiation known to be activated by red light. Here we have used mutants from the two nucleolin genes in Arabidopsis (NUC1 and NUC2) encoding the main regulator of the ribosome biogenesis in the nucleolus in order to better understand their role in adaptive response mechanisms to stress. Thus we show that nucleolin stress-related gene NUC2 can compensate the environmental stress provided by darkness in nuc1 plants while nuc2 plants are not able to provide a complete response to red light. These ground control findings as part of the ESA/NASA Seedling Growth spaceflight experiments will determine the basis for the identification of a genetic background enabling an adaptive advantage for plants in future space experiments.

The Rodent Research-3 (RR-3) mission was sponsored by the pharmaceutical company Eli Lilly and Co. and the Center for the Advancement of Science in Space to study the effectiveness of a potential countermeasure for the loss of muscle and bone mass that occurs during spaceflight. Twenty BALB/c 18-weeks old female mice (ten controls and ten treated) were flown to the ISS and housed in the Rodent Habitat for 39-42 days. Twenty mice of similar age and matching sex and strain were used for ground controls housed in identical hardware and matching ISS environmental conditions. Basal controls were housed in standard vivarium cages. Spaceflight ground controls and basal groups had blood collected then were euthanized had one hind limb removed and finally whole carcasses were stored at -80 C until dissection. All mice in this data set received only the control/sham injection. Brain samples from three flight and three ground control animal groups were cut in half between hemispheres. One hemisphere of each brain was used for generating spatially resolved transcriptional profiling data. Hemispheres were cryosectioned so that 2 consecutive sections from the hippocampus of each brain was placed on Visium Gene Expression arrays. Samples were fixed stained with Hematoxylin and Eosin and imaged. Imaging was followed by tissue permeabilization to release mRNA molecules from cells for capture onto the array surface. Subsequently following the 10XGenomics Visium Gene Expression protocol Spatial Transcriptomics RNA-seq libraries were prepared and sequenced.

The first global genomic proteomic and secondary metabolomic characterization of the filamentous fungus Aspergillus nidulans following growth onboard the International Space Station (ISS) is reported. The investigation included the A. nidulans wild-type and three mutant strains two of which were genetically engineered to enhance secondary metabolite production. Whole genome sequencing revealed that ISS conditions altered the A. nidulans genome in specific regions. In strain CW12001 which features overexpression of the secondary metabolite global regulator laeA ISS conditions induced the loss of the laeA stop codon. Differential expression of proteins involved in stress response carbohydrate metabolic processes and secondary metabolite biosynthesis was also observed. ISS conditions significantly decreased prenyl xanthone production in the wild-type strain and increased asperthecin production in LO1362 and CW12001 which are deficient in a major DNA repair mechanism. These data provide valuable insights into the adaptation mechanism of A. nidulans to spacecraft environments.

Biological risks associated with space radiation and microgravity are major concerns for long-term space travel. Through a Systems Biology approach our previous NASA work has shown both TGF xce xb2 signaling pathways and miRNAs have a critical impact on defining health risks with and without space irradiation. We hypothesize that circulating microRNA (miRNA) signatures are driving microvascular disease and muscle degeneration associated with accelerating aging and will be enhanced by exposure to the space environment (radiation and microgravity). We investigated this hypothesis both in vivo and in vitro and test novel antagonist therapies to these miRNA signatures as countermeasures to reduce space radiation-induced health risks. A comprehensive Systems Biology approach was used to examine the influence by high atomic number by high (H) atomic number (Z) and energy (E) (HZE) irradiation. To simulate low-dose exposure due to galactic cosmic rays (GCR) we used the ions energy and doses determined by a NASA consensus formula of 7 different ions to represent GCR (referred to as GCR sim model). To simulate high-dose radiation exposure due to solar particle events (SPE) we used an acute dose of SPE simulated beam at 1Gy which has energies ranging from 50MeV to 150MeV. C57BL/6 wild-type mice were utilized for irradiation with our established simulated microgravity model (hindlimb suspension model) and an in vitro 3D microvasculature tissue model under simulated microgravity (clinostat) conditions will also be irradiated. To expand on the circulating miRNA signature determined from our preliminary data we determined a group of conserved miRNAs which are commonly being regulated in the majority of the organs and tissues throughout the host using our established techniques. MiRNA-sequencing on serum (before IR and at time of sacrifice) liver heart and muscle tissue for all radiation groups revealed the key circulating miRNA signature (consisting of multiple miRNAs) impacting disease risk. Collectively understanding of how whole body space radiation impacts microvascular and tissue degeneration through circulating miRNAs will greatly enhance health risk prognostication and provide possible new mechanisms for protection against space radiation.

In the Rocket Science citizen science project Eruca sativa (salad rocket) seeds stored for six months on board of the International Space Station caused delayed crop establishment. Here we investigated the physiological and molecular mechanisms underpinning the spaceflight effects on dry seeds. We found that Space seed germination vigor was reduced and aging sensitivity increased but the spaceflight did not compromise seed viability and the development of normal seedlings. Comparative analysis of the transcriptomes (using RNASeq) in dry seeds and upon controlled artificial aging treatment (CAAT) revealed differentially expressed genes (DEGs) associated with spaceflight and ageing. DEG categories enriched by spaceflight and CAAT included transcription and translation with reduced transcript abundances for 40S and 60S ribosomal subunit genes. Among the spaceflight-up DEGs were Heat Shock Protein (HSP) DNAJ-related chaperones a Heat Shock Factor (HSFA7a-like) and components of several DNA repair pathways (e.g. ATM DNA ligase1). The response to radiation category was especially enriched in spaceflight-up DEGs including HSPs catalases and the transcription factor HY5. The major finding from the physiological and transcriptome analysis is that spaceflight causes vigor loss and partial ageing during air-dry seed storage for which space environmental factors and consequences for seed storage during spaceflights are discussed.

The Space Algae Experiment Verification Test completed a competitive growth selection on mutagenized microalgae in a series of batch cultures that were passaged for three growth cycles. The experiment was conducted with ground control conditions similar to those actually used in spaceflight. Cultures were limited in growth rate due to gas permeable membranes to provide oxygen and carbon dioxide exchange and a lack of agitation to mix the cells throughout the liquid media. Two strains were grown and three biological replicates were completed for each strain. Specific variables (factors) tested were: 1) The effect of UVC mutagenesis was tested by sampling the algae cultures prior to mutagenesis. 2) The sensitivity of different strains was tested by conducting the experiment with a wild-type and cell wall mutant (cw15) strain. 3) The effect of competitive growth was tested by sampling each biological replicate experiment at the end of each growth cycle. 4) The effect of live culture storage in the dark was tested by sampling each growth cycle twice. At the time of passage a sample of cells was pelleted and frozen. The cultures were then stored in the dark in a Cargo Transport Bag (CTB) to simulate storage on the ISS and return of cultures. This factor tested whether cultures could be stored alive in the dark or if samples needed to be fixed at the time of passage in order to get an accurate representation of the genetic variation in each cycle of growth. Paired-end whole genome sequencing was completed for 38 samples: 2 strains pre mutagenesis and 2 strains X 3 biological replicates X 3 growth cycles X 2 storage conditions.

The BRIC-21 mission was designed to identify the response of Bacillus subtilis to the human spaceflight environment. For this mission samples were grown in rich-medium using the Biological Research in Canister Petri Dish Fixation Units (BRIC-PDFU) spaceflight hardware. B. subtilis spores were inoculated during spaceflight grown at the ambient ISS temperature and frozen in the onboard -80 C freezer prior to returning to Earth. RNA was extracted from samples grown onboard the International Space Station (ISS) and matching Ground Controls for transcriptome analysis.