{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002032","ANALYSIS_ID":"AN003303","VERSION":"1","CREATED_ON":"January 3, 2022, 7:50 am"},

"PROJECT":{"PROJECT_TITLE":"Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs","PROJECT_SUMMARY":"Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.","INSTITUTE":"University of Colorado Anschutz School of Medicine","LAST_NAME":"Roy","FIRST_NAME":"Micaela","ADDRESS":"1635 Aurora Ct, Aurora, CO, 80045, USA","EMAIL":"micaela.roy@cuanschutz.edu","PHONE":"303-724-3339"},

"STUDY":{"STUDY_TITLE":"Irradiation causes alterations of polyamine, purine and sulfur metabolism in red blood cells and multiple organs (Blood plasma)","STUDY_SUMMARY":"Investigating the metabolic effects of radiation is critical to understand the impact of radiotherapy (e.g., for bone marrow irradiation prior to hematopoietic stem cell transplantation in the clinic or in laboratory studies), space travel, and exposure to environmental radiation. In patients undergoing hemopoietic stem cell transplantation, iron overload is a common risk factor for poor outcomes. Previous studies assert that both irradiation and iron independently modulate tryptophan and indole metabolism of the microbiome, which may in turn impact host immune response. However, no studies have interrogated the multi-organ effects of these treatments concurrently. Herein, we use a model that recapitulate transfusional iron overload, a condition often observed in chronically transfused patients with thalassemia, sickle cell disease, or myelodysplastic syndrome. We applied an omics approach to investigate the impact of both iron load and irradiation on the host metabolome. Our results revealed dose-dependent effects of irradiation in red blood cells (RBC), plasma, spleen, and liver energy and redox metabolism. Increases in polyamines and purine salvage metabolites were observed in organs with high oxygen consumption including the heart, kidney, and brain. Irradiation also impacted the metabolism of the duodenum, colon, and stool, suggesting a potential effect on the microbiome. Iron infusion affected the respose to radiation in the organs and blood, especially in RBC polyamine metabolism and spleen antioxidant metabolism, and affected glucose, sulfur (especially methionine and glutathione systems) and tryptophan metabolism in the liver, stool, and brain. Together, the results suggest that radiation impacts metabolism on a multi-organ level with a significant interaction of host iron status.","INSTITUTE":"University of Colorado Anschutz School of Medicine","LAST_NAME":"Roy","FIRST_NAME":"Micaela","ADDRESS":"1635 Aurora Ct, Aurora, CO, 80045, USA","EMAIL":"micaela.roy@cuanschutz.edu","PHONE":"303-724-3339"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"1",
"Factors":{"Treatment":"Saline","Radiation Dose":"no irradiated"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-1"}
},
{
"Subject ID":"-",
"Sample ID":"2",
"Factors":{"Treatment":"Saline","Radiation Dose":"no irradiated"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-2"}
},
{
"Subject ID":"-",
"Sample ID":"3",
"Factors":{"Treatment":"Saline","Radiation Dose":"no irradiated"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-3"}
},
{
"Subject ID":"-",
"Sample ID":"4",
"Factors":{"Treatment":"Saline","Radiation Dose":"no irradiated"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-4"}
},
{
"Subject ID":"-",
"Sample ID":"6",
"Factors":{"Treatment":"IV iron","Radiation Dose":"no irradiated"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-5"}
},
{
"Subject ID":"-",
"Sample ID":"7",
"Factors":{"Treatment":"IV iron","Radiation Dose":"no irradiated"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-6"}
},
{
"Subject ID":"-",
"Sample ID":"8",
"Factors":{"Treatment":"IV iron","Radiation Dose":"no irradiated"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-7"}
},
{
"Subject ID":"-",
"Sample ID":"9",
"Factors":{"Treatment":"IV iron","Radiation Dose":"no irradiated"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-8"}
},
{
"Subject ID":"-",
"Sample ID":"10",
"Factors":{"Treatment":"IV iron","Radiation Dose":"no irradiated"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-9"}
},
{
"Subject ID":"-",
"Sample ID":"11",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy7"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-10"}
},
{
"Subject ID":"-",
"Sample ID":"13",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy7"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-11"}
},
{
"Subject ID":"-",
"Sample ID":"14",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy7"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-12"}
},
{
"Subject ID":"-",
"Sample ID":"15",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy7"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-13"}
},
{
"Subject ID":"-",
"Sample ID":"16",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy7"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-14"}
},
{
"Subject ID":"-",
"Sample ID":"17",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy7"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-15"}
},
{
"Subject ID":"-",
"Sample ID":"18",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy7"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-16"}
},
{
"Subject ID":"-",
"Sample ID":"19",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy7"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-17"}
},
{
"Subject ID":"-",
"Sample ID":"20",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy7"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-18"}
},
{
"Subject ID":"-",
"Sample ID":"21",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-19"}
},
{
"Subject ID":"-",
"Sample ID":"22",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-20"}
},
{
"Subject ID":"-",
"Sample ID":"23",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-21"}
},
{
"Subject ID":"-",
"Sample ID":"24",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-22"}
},
{
"Subject ID":"-",
"Sample ID":"25",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-23"}
},
{
"Subject ID":"-",
"Sample ID":"26",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-24"}
},
{
"Subject ID":"-",
"Sample ID":"27",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-25"}
},
{
"Subject ID":"-",
"Sample ID":"28",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-26"}
},
{
"Subject ID":"-",
"Sample ID":"29",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-27"}
},
{
"Subject ID":"-",
"Sample ID":"30",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy8"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-28"}
},
{
"Subject ID":"-",
"Sample ID":"31",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-29"}
},
{
"Subject ID":"-",
"Sample ID":"32",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-30"}
},
{
"Subject ID":"-",
"Sample ID":"33",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-31"}
},
{
"Subject ID":"-",
"Sample ID":"34",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-32"}
},
{
"Subject ID":"-",
"Sample ID":"35",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-33"}
},
{
"Subject ID":"-",
"Sample ID":"36",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-34"}
},
{
"Subject ID":"-",
"Sample ID":"37",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-35"}
},
{
"Subject ID":"-",
"Sample ID":"38",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-36"}
},
{
"Subject ID":"-",
"Sample ID":"39",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-37"}
},
{
"Subject ID":"-",
"Sample ID":"40",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy9"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-38"}
},
{
"Subject ID":"-",
"Sample ID":"41",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-39"}
},
{
"Subject ID":"-",
"Sample ID":"42",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-40"}
},
{
"Subject ID":"-",
"Sample ID":"43",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-41"}
},
{
"Subject ID":"-",
"Sample ID":"44",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-42"}
},
{
"Subject ID":"-",
"Sample ID":"45",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-43"}
},
{
"Subject ID":"-",
"Sample ID":"46",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-44"}
},
{
"Subject ID":"-",
"Sample ID":"47",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-45"}
},
{
"Subject ID":"-",
"Sample ID":"48",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-46"}
},
{
"Subject ID":"-",
"Sample ID":"49",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-47"}
},
{
"Subject ID":"-",
"Sample ID":"50",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy10"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-48"}
},
{
"Subject ID":"-",
"Sample ID":"51",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-49"}
},
{
"Subject ID":"-",
"Sample ID":"52",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-50"}
},
{
"Subject ID":"-",
"Sample ID":"53",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-51"}
},
{
"Subject ID":"-",
"Sample ID":"54",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-52"}
},
{
"Subject ID":"-",
"Sample ID":"55",
"Factors":{"Treatment":"Saline","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-53"}
},
{
"Subject ID":"-",
"Sample ID":"56",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-54"}
},
{
"Subject ID":"-",
"Sample ID":"57",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-55"}
},
{
"Subject ID":"-",
"Sample ID":"58",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-56"}
},
{
"Subject ID":"-",
"Sample ID":"59",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-57"}
},
{
"Subject ID":"-",
"Sample ID":"60",
"Factors":{"Treatment":"IV iron","Radiation Dose":"Gy11"},
"Additional sample data":{"RAW_FILE_NAME":"AM3-58"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"At day +4 post irradiation, mice were euthanized and blood was collected by cardiac puncture in heparinized syringe. Plasma obtained after centrifugation at 4500 g for 10 minutes at +4°C","SAMPLE_TYPE":"Blood (plasma)"},

"TREATMENT":{"TREATMENT_SUMMARY":"After one week of acclimatization in a pathogen-free facility, cohorts of mice were retro-orbitally infused with phosphate buffer saline (PBS) or 12.5 mg of iron dextran (Henry Shein Animal Health, Dublin, OH), twice a week for 2 weeks for a total of 50 mg of iron. After 2 days of rest, mice were then divided in groups and irradiated with 7, 8, 9, 10, 11 Gy of C-137 (n=5 per group). Total dose was split in 2 doses 3 hours apart."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"A volume of 50μl of frozen plasma was extracted in 450μl of methanol:acetonitrile:water (5:3:2, v/v/v).29 After vortexing at 4°C for 30 min, extracts were separated from the protein pellet by centrifugation for 10 min at 10,000g at 4°C and stored at −80°C until analysis."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Phenomenex Kinetex C18 (150 x 2.1mm, 2.6 um)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Samples (10uL injection for cells, 20uL injection for SUPs) were introduced to the MS via electrospray ionization with the MS scanning in full MS mode (2 µscans) and ddMS2 (top15) over the range of 65-950 m/z. Technical replicates were injected every six to twelve samples to ensure instrument stability. Metabolites were manually selected integrated with Maven (Princeton University) in conjunction with the KEGG database. Peak quality was determined using blanks, technical mixes, and 13C abundance."},

"MS_METABOLITE_DATA":{
"Units":"peak area top",

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ID":"C00319","parent":"300.2891","medRt":"1.98"},{"Metabolite":"Sphingosine 1-phosphate","KEGG ID":"C06124","parent":"380.2553","medRt":"1.90"},{"Metabolite":"Acetylcholine","KEGG ID":"C01996","parent":"146.1175","medRt":"0.67"},{"Metabolite":"L-Carnitine","KEGG ID":"C00318","parent":"162.1123","medRt":"0.66"},{"Metabolite":"acetyl-carnitine (acyl-C2)","KEGG ID":"C02571","parent":"204.1229","medRt":"0.67"},{"Metabolite":"propionyl-carnitine (acyl-C3)","KEGG ID":"C03017","parent":"218.1385","medRt":"0.67"},{"Metabolite":"butanoyl-l-carnitine (acyl-C4)","KEGG ID":"C02862","parent":"232.1540","medRt":"1.70"},{"Metabolite":"acyl-C4-OH (Hydroxybutyrylcarnitine)","KEGG ID":"HMDB13127","parent":"248.1486","medRt":"0.67"},{"Metabolite":"acyl-C5:1 (Tiglylcarnitine)","KEGG ID":"HMDB02366","parent":"244.1540","medRt":"1.92"},{"Metabolite":"acyl-C5-OH (hydroxyisovalerylcarnitine)","KEGG ID":"ac107","parent":"262.1642","medRt":"0.67"},{"Metabolite":"hexanoyl-L-carnitine (acyl-C6)","KEGG ID":"HMDB00756","parent":"260.1854","medRt":"1.77"},{"Metabolite":"L-octanoylcarnitine (acyl-C8)","KEGG ID":"C02838","parent":"288.2163","medRt":"1.83"},{"Metabolite":"O-Decenoyl-L-carnitine (acyl-C10:1)","KEGG ID":"CID57357170","parent":"314.2321","medRt":"1.85"},{"Metabolite":"O-dodecanoyl-carnitine (acyl-C12)","KEGG ID":"HMDB02250","parent":"344.2791","medRt":"1.93"},{"Metabolite":"O-dodecenoyl-carnitine (acyl-C12:1)","KEGG ID":"HMDB13326","parent":"342.2635","medRt":"1.91"},{"Metabolite":"O-tetradecanoyl-L-carnitine (acyl-C14)","KEGG ID":"HMDB05066","parent":"372.3103","medRt":"1.99"},{"Metabolite":"Tetradecenoyl Carnitine (acyl-C14:1)","KEGG ID":"HMDB13329","parent":"370.2948","medRt":"1.96"},{"Metabolite":"L-Palmitoylcarnitine (acyl-C16)","KEGG ID":"C02990","parent":"400.3417","medRt":"2.06"},{"Metabolite":"Hexadecenoyl-carnitine (acyl-C16:1)","KEGG ID":"HMDB06317","parent":"398.3260","medRt":"2.00"},{"Metabolite":"O-octadecenoyl-L-carnitine (acyl-C18:1)","KEGG ID":"HMDB06351","parent":"426.3573","medRt":"2.07"},{"Metabolite":"acyl-C18:2 (Linoleoyl-CoA)","KEGG ID":"HMDB01064","parent":"424.3414","medRt":"2.02"},{"Metabolite":"acyl-C18:2-OH","KEGG ID":"ac123","parent":"440.3362","medRt":"1.97"},{"Metabolite":"acyl-C20:4","KEGG ID":"ac125","parent":"448.3408","medRt":"2.01"}]
}

}