#METABOLOMICS WORKBENCH epannkuk_20210723_075834_mwtab.txt DATATRACK_ID:2766 STUDY_ID:ST001891 ANALYSIS_ID:AN003071 PROJECT_ID:PR001191
VERSION             	1
CREATED_ON             	August 2, 2021, 12:17 pm
#PROJECT
PR:PROJECT_TITLE                 	Small molecule signatures of mice lacking T-cell p38 alternate activation, a
PR:PROJECT_TITLE                 	model for immunosuppression conditions, after exposure to total body radiation
PR:PROJECT_SUMMARY               	Introduction Novel biodosimetry assays are needed in the event of
PR:PROJECT_SUMMARY               	radiological/nuclear emergencies for both immediate triage and identifying
PR:PROJECT_SUMMARY               	delayed effects of acute radiation exposure. Genetically engineered mouse models
PR:PROJECT_SUMMARY               	are used to assess how genotypic variation in the general population may affect
PR:PROJECT_SUMMARY               	post-irradiation classification performance. Here, we used a mouse model that
PR:PROJECT_SUMMARY               	lacks the T-cell receptor specific alternative p38 pathway (p38αβY323F, double
PR:PROJECT_SUMMARY               	knock-in [DKI] mice) to determine how attenuated autoimmune and inflammatory
PR:PROJECT_SUMMARY               	responses may affect dose reconstruction. Objectives To determine if deficient
PR:PROJECT_SUMMARY               	alternative p38 activation differentially affects biofluid metabolic signatures
PR:PROJECT_SUMMARY               	post-irradiation compared to wild-type (WT). Methods Untargeted global
PR:PROJECT_SUMMARY               	metabolomics was used to assess biofluid signatures between WT and DKI mice (8
PR:PROJECT_SUMMARY               	– 10 weeks old) after exposure to total body radiation (0, 2, or 7 Gy). Urine
PR:PROJECT_SUMMARY               	was analyzed in the first week (1, 3, and 7 d) and serum at 1 d. Spectral
PR:PROJECT_SUMMARY               	features of interest were identified using the machine learning algorithm Random
PR:PROJECT_SUMMARY               	Forests and MetaboLyzer. Validated metabolite panels were constructed and
PR:PROJECT_SUMMARY               	classification performance was assessed by determining the area under the
PR:PROJECT_SUMMARY               	receiver operating characteristic curve (AUROC). Results A multidimensional
PR:PROJECT_SUMMARY               	scaling plot showed excellent separation of IR exposed groups in WT with
PR:PROJECT_SUMMARY               	slightly dampened responses in DKI mice. For both urine and serum, excellent
PR:PROJECT_SUMMARY               	sensitivity and specificity (AUROC > 0.90) was observed for 0 Gy vs. 7 Gy groups
PR:PROJECT_SUMMARY               	irrespective of genotype using identical metabolite panels. Similarly, excellent
PR:PROJECT_SUMMARY               	to fair classification (AUROC > 0.75) was observed for ≤ 2 Gy vs. 7 Gy
PR:PROJECT_SUMMARY               	post-irradiation mice for both genotypes, however, model performance declined
PR:PROJECT_SUMMARY               	(AUROC < 0.75) between genotypes post-irradiation. Conclusion Overall, these
PR:PROJECT_SUMMARY               	results suggest less influence of the alternative p38 activation pathway for
PR:PROJECT_SUMMARY               	dose reconstruction compared to other radiosensitive genotypes.
PR:INSTITUTE                     	Georgetown University
PR:LAST_NAME                     	Pannkuk
PR:FIRST_NAME                    	Evan
PR:ADDRESS                       	3970 Reservoir Rd, NW New Research Building E504
PR:EMAIL                         	elp44@georgetown.edu
PR:PHONE                         	2026875650
#STUDY
ST:STUDY_TITLE                   	Small molecule signatures of mice lacking T-cell p38 alternate activation, a
ST:STUDY_TITLE                   	model for immunosuppression conditions, after exposure to total body radiation
ST:STUDY_TITLE                   	(part I)
ST:STUDY_SUMMARY                 	Introduction Novel biodosimetry assays are needed in the event of
ST:STUDY_SUMMARY                 	radiological/nuclear emergencies for both immediate triage and identifying
ST:STUDY_SUMMARY                 	delayed effects of acute radiation exposure. Genetically engineered mouse models
ST:STUDY_SUMMARY                 	are used to assess how genotypic variation in the general population may affect
ST:STUDY_SUMMARY                 	post-irradiation classification performance. Here, we used a mouse model that
ST:STUDY_SUMMARY                 	lacks the T-cell receptor specific alternative p38 pathway (p38αβY323F, double
ST:STUDY_SUMMARY                 	knock-in [DKI] mice) to determine how attenuated autoimmune and inflammatory
ST:STUDY_SUMMARY                 	responses may affect dose reconstruction. Objectives To determine if deficient
ST:STUDY_SUMMARY                 	alternative p38 activation differentially affects biofluid metabolic signatures
ST:STUDY_SUMMARY                 	post-irradiation compared to wild-type (WT). Methods Untargeted global
ST:STUDY_SUMMARY                 	metabolomics was used to assess biofluid signatures between WT and DKI mice (8
ST:STUDY_SUMMARY                 	– 10 weeks old) after exposure to total body radiation (0, 2, or 7 Gy). Urine
ST:STUDY_SUMMARY                 	was analyzed in the first week (1, 3, and 7 d) and serum at 1 d. Spectral
ST:STUDY_SUMMARY                 	features of interest were identified using the machine learning algorithm Random
ST:STUDY_SUMMARY                 	Forests and MetaboLyzer. Validated metabolite panels were constructed and
ST:STUDY_SUMMARY                 	classification performance was assessed by determining the area under the
ST:STUDY_SUMMARY                 	receiver operating characteristic curve (AUROC). Results A multidimensional
ST:STUDY_SUMMARY                 	scaling plot showed excellent separation of IR exposed groups in WT with
ST:STUDY_SUMMARY                 	slightly dampened responses in DKI mice. For both urine and serum, excellent
ST:STUDY_SUMMARY                 	sensitivity and specificity (AUROC > 0.90) was observed for 0 Gy vs. 7 Gy groups
ST:STUDY_SUMMARY                 	irrespective of genotype using identical metabolite panels. Similarly, excellent
ST:STUDY_SUMMARY                 	to fair classification (AUROC > 0.75) was observed for ≤ 2 Gy vs. 7 Gy
ST:STUDY_SUMMARY                 	post-irradiation mice for both genotypes, however, model performance declined
ST:STUDY_SUMMARY                 	(AUROC < 0.75) between genotypes post-irradiation. Conclusion Overall, these
ST:STUDY_SUMMARY                 	results suggest less influence of the alternative p38 activation pathway for
ST:STUDY_SUMMARY                 	dose reconstruction compared to other radiosensitive genotypes.
ST:INSTITUTE                     	Georgetown University
ST:LAST_NAME                     	Pannkuk
ST:FIRST_NAME                    	Evan
ST:ADDRESS                       	3970 Reservoir Rd, NW New Research Building E504
ST:EMAIL                         	elp44@georgetown.edu
ST:PHONE                         	2026875650
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
SU:GENDER                        	Male
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	260	9	Irradiation:2Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_015
SUBJECT_SAMPLE_FACTORS           	126	10	Irradiation:7Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_016
SUBJECT_SAMPLE_FACTORS           	183	12	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_019
SUBJECT_SAMPLE_FACTORS           	121	13	Irradiation:2Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_020
SUBJECT_SAMPLE_FACTORS           	123	14	Irradiation:2Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_021
SUBJECT_SAMPLE_FACTORS           	182	15	Irradiation:2Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_022
SUBJECT_SAMPLE_FACTORS           	316	16	Irradiation:2Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_023
SUBJECT_SAMPLE_FACTORS           	316	20	Irradiation:2Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_027
SUBJECT_SAMPLE_FACTORS           	127	21	Irradiation:7Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_029
SUBJECT_SAMPLE_FACTORS           	260	25	Irradiation:2Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_033
SUBJECT_SAMPLE_FACTORS           	183	26	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_034
SUBJECT_SAMPLE_FACTORS           	128	29	Irradiation:7Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_037
SUBJECT_SAMPLE_FACTORS           	130	30	Irradiation:7Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_038
SUBJECT_SAMPLE_FACTORS           	379	31	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_040
SUBJECT_SAMPLE_FACTORS           	726	32	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_041
SUBJECT_SAMPLE_FACTORS           	728	33	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_042
SUBJECT_SAMPLE_FACTORS           	183	34	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_043
SUBJECT_SAMPLE_FACTORS           	317	41	Irradiation:2Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_051
SUBJECT_SAMPLE_FACTORS           	314	42	Irradiation:2Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_052
SUBJECT_SAMPLE_FACTORS           	123	46	Irradiation:2Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_056
SUBJECT_SAMPLE_FACTORS           	260	48	Irradiation:2Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_058
SUBJECT_SAMPLE_FACTORS           	316	49	Irradiation:2Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_059
SUBJECT_SAMPLE_FACTORS           	183	50	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_060
SUBJECT_SAMPLE_FACTORS           	123	51	Irradiation:2Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_062
SUBJECT_SAMPLE_FACTORS           	182	62	Irradiation:2Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_074
SUBJECT_SAMPLE_FACTORS           	314	63	Irradiation:2Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_075
SUBJECT_SAMPLE_FACTORS           	327	64	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_076
SUBJECT_SAMPLE_FACTORS           	122	66	Irradiation:2Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_078
SUBJECT_SAMPLE_FACTORS           	125	67	Irradiation:2Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_079
SUBJECT_SAMPLE_FACTORS           	130	68	Irradiation:7Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_080
SUBJECT_SAMPLE_FACTORS           	726	69	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_081
SUBJECT_SAMPLE_FACTORS           	379	72	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_085
SUBJECT_SAMPLE_FACTORS           	122	75	Irradiation:2Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_088
SUBJECT_SAMPLE_FACTORS           	126	80	Irradiation:7Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_093
SUBJECT_SAMPLE_FACTORS           	123	83	Irradiation:2Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_097
SUBJECT_SAMPLE_FACTORS           	125	87	Irradiation:2Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_101
SUBJECT_SAMPLE_FACTORS           	732	88	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_102
SUBJECT_SAMPLE_FACTORS           	728	90	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_104
SUBJECT_SAMPLE_FACTORS           	182	93	Irradiation:2Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_108
SUBJECT_SAMPLE_FACTORS           	194	94	Irradiation:2Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_109
SUBJECT_SAMPLE_FACTORS           	317	95	Irradiation:2Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_110
SUBJECT_SAMPLE_FACTORS           	314	96	Irradiation:2Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_111
SUBJECT_SAMPLE_FACTORS           	121	99	Irradiation:2Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_114
SUBJECT_SAMPLE_FACTORS           	121	112	Irradiation:2Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_129
SUBJECT_SAMPLE_FACTORS           	326	114	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_131
SUBJECT_SAMPLE_FACTORS           	328	115	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_132
SUBJECT_SAMPLE_FACTORS           	260	116	Irradiation:2Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_133
SUBJECT_SAMPLE_FACTORS           	182	117	Irradiation:2Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_134
SUBJECT_SAMPLE_FACTORS           	194	118	Irradiation:2Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_135
SUBJECT_SAMPLE_FACTORS           	315	119	Irradiation:2Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_136
SUBJECT_SAMPLE_FACTORS           	316	120	Irradiation:2Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_137
SUBJECT_SAMPLE_FACTORS           	327	121	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_139
SUBJECT_SAMPLE_FACTORS           	121	126	Irradiation:2Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_144
SUBJECT_SAMPLE_FACTORS           	124	127	Irradiation:2Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_145
SUBJECT_SAMPLE_FACTORS           	731	128	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_146
SUBJECT_SAMPLE_FACTORS           	125	130	Irradiation:2Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_148
SUBJECT_SAMPLE_FACTORS           	126	131	Irradiation:7Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_150
SUBJECT_SAMPLE_FACTORS           	726	132	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_151
SUBJECT_SAMPLE_FACTORS           	126	137	Irradiation:7Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_156
SUBJECT_SAMPLE_FACTORS           	726	138	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_157
SUBJECT_SAMPLE_FACTORS           	728	139	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_158
SUBJECT_SAMPLE_FACTORS           	127	142	Irradiation:7Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_162
SUBJECT_SAMPLE_FACTORS           	315	145	Irradiation:2Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_165
SUBJECT_SAMPLE_FACTORS           	122	148	Irradiation:2Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_168
SUBJECT_SAMPLE_FACTORS           	129	149	Irradiation:7Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_169
SUBJECT_SAMPLE_FACTORS           	731	150	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_170
SUBJECT_SAMPLE_FACTORS           	317	152	Irradiation:2Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_173
SUBJECT_SAMPLE_FACTORS           	318	153	Irradiation:2Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_174
SUBJECT_SAMPLE_FACTORS           	331	154	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_175
SUBJECT_SAMPLE_FACTORS           	129	158	Irradiation:7Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_179
SUBJECT_SAMPLE_FACTORS           	379	159	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_180
SUBJECT_SAMPLE_FACTORS           	194	163	Irradiation:2Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_185
SUBJECT_SAMPLE_FACTORS           	315	164	Irradiation:2Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_186
SUBJECT_SAMPLE_FACTORS           	324	165	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_187
SUBJECT_SAMPLE_FACTORS           	327	166	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_188
SUBJECT_SAMPLE_FACTORS           	331	167	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_189
SUBJECT_SAMPLE_FACTORS           	130	172	Irradiation:7Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_195
SUBJECT_SAMPLE_FACTORS           	731	173	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_196
SUBJECT_SAMPLE_FACTORS           	315	174	Irradiation:2Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_197
SUBJECT_SAMPLE_FACTORS           	318	175	Irradiation:2Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_198
SUBJECT_SAMPLE_FACTORS           	327	176	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_199
SUBJECT_SAMPLE_FACTORS           	331	177	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_200
SUBJECT_SAMPLE_FACTORS           	124	184	Irradiation:2Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_208
SUBJECT_SAMPLE_FACTORS           	125	185	Irradiation:2Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_209
SUBJECT_SAMPLE_FACTORS           	194	205	Irradiation:2Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_231
SUBJECT_SAMPLE_FACTORS           	318	206	Irradiation:2Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_232
SUBJECT_SAMPLE_FACTORS           	325	207	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_233
SUBJECT_SAMPLE_FACTORS           	184	208	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_234
SUBJECT_SAMPLE_FACTORS           	326	213	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_240
SUBJECT_SAMPLE_FACTORS           	328	214	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_241
SUBJECT_SAMPLE_FACTORS           	128	221	Irradiation:7Gy | Genotype:WT | Collection_time:1d	RAW_FILE_NAME=POS_249
SUBJECT_SAMPLE_FACTORS           	732	222	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_250
SUBJECT_SAMPLE_FACTORS           	326	223	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_251
SUBJECT_SAMPLE_FACTORS           	127	224	Irradiation:7Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_252
SUBJECT_SAMPLE_FACTORS           	325	226	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_254
SUBJECT_SAMPLE_FACTORS           	732	230	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_258
SUBJECT_SAMPLE_FACTORS           	184	235	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_264
SUBJECT_SAMPLE_FACTORS           	122	236	Irradiation:2Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_265
SUBJECT_SAMPLE_FACTORS           	324	239	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_268
SUBJECT_SAMPLE_FACTORS           	124	244	Irradiation:2Gy | Genotype:WT | Collection_time:pre	RAW_FILE_NAME=POS_274
SUBJECT_SAMPLE_FACTORS           	314	245	Irradiation:2Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_275
SUBJECT_SAMPLE_FACTORS           	324	246	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_276
SUBJECT_SAMPLE_FACTORS           	325	247	Irradiation:7Gy | Genotype:p38 | Collection_time:1d	RAW_FILE_NAME=POS_277
SUBJECT_SAMPLE_FACTORS           	328	254	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_285
SUBJECT_SAMPLE_FACTORS           	732	261	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_293
SUBJECT_SAMPLE_FACTORS           	331	281	Irradiation:7Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_315
SUBJECT_SAMPLE_FACTORS           	328	283	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_317
SUBJECT_SAMPLE_FACTORS           	731	284	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_318
SUBJECT_SAMPLE_FACTORS           	317	285	Irradiation:2Gy | Genotype:p38 | Collection_time:pre	RAW_FILE_NAME=POS_319
SUBJECT_SAMPLE_FACTORS           	325	294	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_329
SUBJECT_SAMPLE_FACTORS           	124	296	Irradiation:2Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_331
SUBJECT_SAMPLE_FACTORS           	129	297	Irradiation:7Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_332
SUBJECT_SAMPLE_FACTORS           	379	301	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_337
SUBJECT_SAMPLE_FACTORS           	318	321	Irradiation:2Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_359
SUBJECT_SAMPLE_FACTORS           	128	327	Irradiation:7Gy | Genotype:WT | Collection_time:3d	RAW_FILE_NAME=POS_365
SUBJECT_SAMPLE_FACTORS           	324	328	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_366
SUBJECT_SAMPLE_FACTORS           	326	329	Irradiation:7Gy | Genotype:p38 | Collection_time:7d	RAW_FILE_NAME=POS_367
SUBJECT_SAMPLE_FACTORS           	128	332	Irradiation:7Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_371
SUBJECT_SAMPLE_FACTORS           	130	333	Irradiation:7Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_372
SUBJECT_SAMPLE_FACTORS           	728	341	Irradiation:7Gy | Genotype:p38 | Collection_time:3d	RAW_FILE_NAME=POS_381
SUBJECT_SAMPLE_FACTORS           	127	346	Irradiation:7Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_386
SUBJECT_SAMPLE_FACTORS           	129	348	Irradiation:7Gy | Genotype:WT | Collection_time:7d	RAW_FILE_NAME=POS_388
#COLLECTION
CO:COLLECTION_SUMMARY            	Urine was collected after irradiation
CO:SAMPLE_TYPE                   	Urine
#TREATMENT
TR:TREATMENT_SUMMARY             	WT C57Bl/6 mice (C57BL/6NCrl strain code #027) were obtained from Charles River
TR:TREATMENT_SUMMARY             	Laboratories (Frederick, MD) and DKI mice were kindly provided by the Laboratory
TR:TREATMENT_SUMMARY             	of Immune Cell Biology, National Cancer Institute (P.I. Jonathan D. Ashwell,
TR:TREATMENT_SUMMARY             	M.D.) (Jirmanova et al. 2011). Animals were bred/irradiated (12 h light / 12 h
TR:TREATMENT_SUMMARY             	dark cycle conditions) at Georgetown University and water and food (PicoLab
TR:TREATMENT_SUMMARY             	Rodent Diet 20 #5053) were provided ad libitum according to Georgetown
TR:TREATMENT_SUMMARY             	University Institutional Animal Care and Use Committee (GUACUC) protocols
TR:TREATMENT_SUMMARY             	(2016-1152). Before irradiation and biofluid collection the mice were acclimated
TR:TREATMENT_SUMMARY             	to metabolic cages for 24 h. Male mice that were 8 – 10 weeks old were exposed
TR:TREATMENT_SUMMARY             	to a total body ionization (TBI) x-ray dose (~1.67 Gy/min; X-Rad 320, Precision
TR:TREATMENT_SUMMARY             	X-Ray Inc, Branford, CT; filter, 0.75 mm tin/ 0.25 mm copper/1.5 mm aluminum) of
TR:TREATMENT_SUMMARY             	0, 2, or 7 Gy. All urine samples were collected over a 24 h period in a
TR:TREATMENT_SUMMARY             	metabolic cage pre-irradiation and at days 1, 3, and 7 d post-irradiation
TR:TREATMENT_SUMMARY             	(Figure S1). Blood for metabolomics was collected at 1 d via cheek bleed from
TR:TREATMENT_SUMMARY             	the submandibular vein and serum was separated in a BD microtainer serum
TR:TREATMENT_SUMMARY             	separator tube and centrifuged for 10 min (10,000 x g, 4°C). Serum samples from
TR:TREATMENT_SUMMARY             	sham-irradiated mice were used as a control (Figure S1). All biofluids were
TR:TREATMENT_SUMMARY             	flash frozen and stored at -80°C until further use. Seven days
TR:TREATMENT_SUMMARY             	post-irradiation, blood was collected in a dipotassium EDTA Tube (BD Cat
TR:TREATMENT_SUMMARY             	#365974) via the facial vein from each animal and subjected to a complete blood
TR:TREATMENT_SUMMARY             	count by VRL Diagnostics (Gaithersburg, MD, http://www.vrlsat.com/) (Figure S2).
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Biofluids were prepared as previously described (Pannkuk et al. 2018;2020).
SP:SAMPLEPREP_SUMMARY            	Urine (20 μl) was deproteinated with 50% acetonitrile (80 μl) containing
SP:SAMPLEPREP_SUMMARY            	internal standards (2 μM debrisoquine sulfate, 30 μM 4-nitrobenzoic acid),
SP:SAMPLEPREP_SUMMARY            	incubated on ice for 10 min, vortexed for 30 seconds, and centrifuged for 10 min
SP:SAMPLEPREP_SUMMARY            	(10,000 x g, 4°C). Serum (5 μl) was prepared as above but was deproteinated
SP:SAMPLEPREP_SUMMARY            	with 66% acetonitrile (195 μl). A quality control (QC) sample was prepared by
SP:SAMPLEPREP_SUMMARY            	mixing 1 μl from each sample and prepared as above.
SP:PROCESSING_STORAGE_CONDITIONS 	-80℃
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Mobile phases consisted of the following: solvent A (water/0.1% formic acid
CH:CHROMATOGRAPHY_SUMMARY        	[FA]), solvent B (ACN/0.1% FA), solvent C (isopropanol [IPA]/ACN (90:10)/0.1%
CH:CHROMATOGRAPHY_SUMMARY        	FA). The gradient for urine was (solvent A and B) 4.0 min 5% B, 4.0 min 20% B,
CH:CHROMATOGRAPHY_SUMMARY        	5.1 min 95% B, and 1.9 min 5% B at a flow rate of 0.5 ml/min. The gradient for
CH:CHROMATOGRAPHY_SUMMARY        	serum was (solvent A, B, and C) 4.0 min 98:2 A:B, 4.0 min 40:60 A:B, 1.5 min
CH:CHROMATOGRAPHY_SUMMARY        	2:98 A:B, 2.0 min 2:98 A:C, 0.5 min 50:50 A:C, and 1.0 min 98:2 A:B at a flow
CH:CHROMATOGRAPHY_SUMMARY        	rate of 0.5 ml/min.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Waters Acquity
CH:COLUMN_NAME                   	Waters Acquity BEH C18 (50 x 2.1mm, 1.7 um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Waters Synapt G2 QTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Negative and positive electrospray ionization (ESI) data-independent modes were
MS:MS_COMMENTS                   	used for data acquisition with leucine enkephalin ([M+H]+ = 556.2771, [M-H]- =
MS:MS_COMMENTS                   	554.2615) as Lock-Spray®. Operating conditions for ESI were: capillary voltage
MS:MS_COMMENTS                   	2.75 kV, cone voltage 30 V, desolvation temperature 500°C, desolvation gas flow
MS:MS_COMMENTS                   	1000 L/Hr.
MS:MS_RESULTS_FILE               	ST001891_AN003071_Results.txt	UNITS:peak area	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END