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We also thank the Biomedical Imaging Core at the UA College of Medicine - Phoenix for providing epifluorescent imaging services. We express our gratitude to Drs. Christopher D. Porada, Matthew A. Coleman, and Paul F. Wilson for bringing this study together and for their tremendous help at the NSRL facilities. We also extend our thanks to all the staff at Brookhaven National Laboratory, and in particular to the team at NSRL, for coordinating irradiation and helping with shipments. Finally, we would like to thank Dr. Paul Wilmes for his initial discussions and expertise in microbiology, which helped shape this project."},{"name":"DOI","value":"10.26030/y1ww-ep94"},{"name":"Data Source Accession","value":""},{"name":"Data Source Link","value":""},{"name":"Experiment Platform","value":""},{"name":"Flight Program","value":""},{"name":"Funding","value":"This study was mostly supported by the NASA Translational Research Institute for Space Health (TRISH) through Cooperative Agreement NNX16AO69A."},{"name":"Identifiers","value":""},{"name":"Managing NASA Center","value":""},{"name":"Mission End","value":""},{"name":"Mission Link","value":""},{"name":"Mission Name","value":""},{"name":"Mission Start","value":""},{"name":"Project Identifier","value":"High_LET_Irradiation, Multiple_Radiation_Types, High_Dose_Irradiation"},{"name":"Project Link","value":""},{"name":"Project Title","value":""},{"name":"Project Type","value":"Ground Study"},{"name":"Space Program","value":"NASA"},{"name":"Study Grant Number","value":""}],"description":"Space radiation poses significant risks to human health, particularly to the radiation-sensitive gastrointestinal tract, yet the role of the microbiome in modulating intestinal responses to space-relevant heavy-ion exposure remains poorly understood due to the lack of advanced human-relevant experimental models. Here, we adapted a previously developed gut-on-chip (HuMiX) that recapitulates host-microbiome crosstalk to investigate the specific role of Lacticaseibacillus rhamnosus GG (LGG) on the radiation response of normal human intestinal cells (FHs 74 Int) after exposure to 600 MeV per nucleon Si-28 or 1 GeV per nucleon Fe-56 ions. First, results showed that LGG induced transcriptional reprogramming characterized by reduced proliferative signaling and changes in metabolic and epithelial organization pathway, confirming efficient interaction with the FHs 74 Int cells. Irradiation at an absorbed dose of 50 cGy of Si-28 or Fe-56 ions triggered canonical radiation responses, including activation of inflammatory signaling, DNA damage pathways, p53 signaling, and cell-cycle checkpoints, while suppressing developmental and epithelial differentiation programs. Despite shared radiation-response signatures, the two radiation qualities produced distinct transcriptional profiles: Fe-56 preferentially activated inflammatory and lipid metabolic pathways, whereas Si-28 induced interferon signaling, oxidative phosphorylation, and translation-related programs. Importantly, LGG modulated the radiation response with differences in DNA repair- and replication-associated transcriptional programs. Quantification revealed significantly reduced residual gamma-H2AX-53BP1 DNA damage foci in irradiated LGG-exposed cells compared with irradiated cells cultured without LGG. Together, these findings demonstrate the utility of organ-on-chip platforms to investigate human tissue responses to space radiation and reveal a potential microbiome-mediated mechanism of intestinal radioprotection.","factors":[{"@id":"HdeENB9q76O15DqwRnzSdxiccxgsmRU9MSFssmJ9","comments":[],"factorName":"Ionizing radiation","factorType":{"@id":"lxbLs9W1zXxYT4z2FHb8baoCdWPNdwXmQcARkZcI","annotationValue":"ionizing radiation","comments":[],"termAccession":"http://purl.obolibrary.org/obo/RBO_00000116","termSource":"RBO"}},{"@id":"ptDUh5ykW4PxMIVIzODauUcAICOxDYBCkCMYN6kl","comments":[],"factorName":"Culture Platform","factorType":{"@id":"ZspxAnFx5bNo3Xihv4t7Fn9N75SPCNTacoqOBqbj","annotationValue":"culture 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Adapter-ligated libraries were then PCR-amplified and sequenced on an Illumina platform.","name":"Library Construction","parameters":[{"@id":"#139726702505456","parameterName":{"@id":"SXpkstgzgf9WOhn8BRUImqf5kbHf1TJ3GjkLiaMg","annotationValue":"Library Selection","comments":[],"termAccession":"","termSource":""}},{"@id":"#139726702505744","parameterName":{"@id":"rlzTK2chbZkUcaCWHWpzFmlVUzKNZ7DXWAzUXMTb","annotationValue":"Library Layout","comments":[],"termAccession":"","termSource":""}}],"protocolType":{"@id":"pzj9RZv3CsjT04c70GAAlKfarf7MTNECAkwnLMzP","annotationValue":"Library Construction","comments":[],"termAccession":"","termSource":"RBO"},"uri":"","version":""},{"@id":"#139726645445392","comments":[],"components":[],"description":"RNA-seq was performed by Azenta Life Sciences using an RNA-seq package including polyA selection and sequencing using an Illumina HiSeq for 150 bp paired-end reads.","name":"Nucleic Acid Sequencing","parameters":[{"@id":"#139726702504880","parameterName":{"@id":"QZv3O6FfjrX7Rj1AdVBpE7MMkvX0gDPFNyhTgsul","annotationValue":"Sequencing Instrument","comments":[],"termAccession":"","termSource":""}},{"@id":"#139726702503584","parameterName":{"@id":"Gpt4vDjDrU84g95ETwLXCbrKXjfl1JbGRu6ncg88","annotationValue":"Read Length","comments":[],"termAccession":"","termSource":""}},{"@id":"#139726702505696","parameterName":{"@id":"djbnytZ0pcYQzJLwtVqtWq5iA8gbrVJIALsL2zf6","annotationValue":"Raw Data File","comments":[],"termAccession":"https://osdr.nasa.gov/","termSource":"OSD"}}],"protocolType":{"@id":"QBkRk9iAy6j4WJ0qjnDo431vZDjQH5gO6PXf5OEK","annotationValue":"Nucleic Acid Sequencing","comments":[],"termAccession":"","termSource":"RBO"},"uri":"","version":""},{"@id":"#139726645445104","comments":[],"components":[],"description":"The sequence reads were trimmed using Trimmomatic v.0.36 to eliminate adapter sequences and nucleotides with poor quality. 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