#METABOLOMICS WORKBENCH tara600_20221205_170349 DATATRACK_ID:3628 STUDY_ID:ST002390 ANALYSIS_ID:AN003894 VERSION 1 CREATED_ON 02-08-2024 #PROJECT PR:PROJECT_TITLE The metabolomic resetting effect of DMXAA in cisplatin-induced AKI PR:PROJECT_TYPE MS quantitative analysis PR:PROJECT_SUMMARY Cisplatin-induced nephrotoxicity is the main adverse effect of cisplatin-based PR:PROJECT_SUMMARY chemotherapy, which highly limits the clinical use of cisplatin. DMXAA, a PR:PROJECT_SUMMARY flavonoid derivative as a known agonist of STING, has been served as a promising PR:PROJECT_SUMMARY antivascular agent. Although cGAS-STING activation has been demonstrated to PR:PROJECT_SUMMARY mediate cisplatin-induced AKI, the role of DMXAA in this condition is unclear. PR:PROJECT_SUMMARY Here, we defined an unexpected and critical role of DMXAA in improving renal PR:PROJECT_SUMMARY function, ameliorating renal tubular injury and cell apoptosis, suppressing PR:PROJECT_SUMMARY inflammation in cisplatin-induced AKI. Moreover, we confirmed that DMXAA PR:PROJECT_SUMMARY combated AKI in a STING-independent manner evidenced by its protection against PR:PROJECT_SUMMARY AKI in STING knockout mice. Due to the established role of metabolic disorders PR:PROJECT_SUMMARY in AKI, which could contribute to the injury and kidney recovery, we performed PR:PROJECT_SUMMARY metabolomics using renal tissues from cisplatin-induced AKI mice with or without PR:PROJECT_SUMMARY DMXAA treatment. Strikingly, the date revealed that DMXAA improved the metabolic PR:PROJECT_SUMMARY disorders in kidneys of AKI mice, especially restored the tryptophan metabolism. PR:INSTITUTE Children's Hospital of Nanjing Medical University PR:DEPARTMENT Department of Nephrology, State Key Laboratory of Reproductive Medicine, PR:LABORATORY Nanjing Key Lab of Pediatrics, Jiangsu Key Laboratory of Pediatrics PR:LAST_NAME Lu PR:FIRST_NAME Lingling PR:ADDRESS Guangzhou Road 72, Nanjing, Jiangsu, 210000, China PR:EMAIL lulingling89tara@163.com PR:PHONE 0086-25-8311-7435 PR:DOI http://dx.doi.org/10.21228/M81B0N #STUDY ST:STUDY_TITLE The metabolomic resetting effect of DMXAA in cisplatin-induced injured mouse ST:STUDY_TITLE kidney ST:STUDY_SUMMARY A single injection of cisplatin was used to induce AKI in mice. To assess the ST:STUDY_SUMMARY effect of DMXAA, the mice were injected intraperitoneally with DMXAA or vehicle ST:STUDY_SUMMARY one hour before cisplatin injection, repeated every 24 hours. The mice were ST:STUDY_SUMMARY euthanized 72 hours after cisplatin injection, and renal tissues were collected ST:STUDY_SUMMARY for MS analysis. ST:INSTITUTE Children's Hospital of Nanjing Medical University ST:LAST_NAME Lu ST:FIRST_NAME Lingling ST:ADDRESS Guangzhou Road 72, Nanjing, Jiangsu, 210000, China ST:EMAIL lulingling89tara@163.com ST:PHONE 0086-25-8311-7435 ST:SUBMIT_DATE 2022-12-05 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN C57BL/6J, wild type SU:AGE_OR_AGE_RANGE 8 weeks SU:GENDER Male #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - C_1 Treatment:control Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_C_1.mzML; RAW_FILE_NAME=neg_C_1.mzML SUBJECT_SAMPLE_FACTORS - C_2 Treatment:control Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_C_2.mzML; RAW_FILE_NAME=neg_C_2.mzML SUBJECT_SAMPLE_FACTORS - C_3 Treatment:control Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_C_3.mzML; RAW_FILE_NAME=neg_C_3.mzML SUBJECT_SAMPLE_FACTORS - C_4 Treatment:control Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_C_4.mzML; RAW_FILE_NAME=neg_C_4.mzML SUBJECT_SAMPLE_FACTORS - C_5 Treatment:control Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_C_5.mzML; RAW_FILE_NAME=neg_C_5.mzML SUBJECT_SAMPLE_FACTORS - XP_1 Treatment:DMXAA+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_XP_1.mzML; RAW_FILE_NAME=neg_XP_1.mzML SUBJECT_SAMPLE_FACTORS - X_P_2 Treatment:DMXAA+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_XP_2.mzML; RAW_FILE_NAME=neg_XP_2.mzML SUBJECT_SAMPLE_FACTORS - X_P_3 Treatment:DMXAA+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_XP_3.mzML; RAW_FILE_NAME=neg_XP_3.mzML SUBJECT_SAMPLE_FACTORS - X_P_4 Treatment:DMXAA+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_XP_4.mzML; RAW_FILE_NAME=neg_XP_4.mzML SUBJECT_SAMPLE_FACTORS - X_P_5 Treatment:DMXAA+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_XP_5.mzML; RAW_FILE_NAME=neg_XP_5.mzML SUBJECT_SAMPLE_FACTORS - P_1 Treatment:vehicle+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_P_1.mzML; RAW_FILE_NAME=neg_P_1.mzML SUBJECT_SAMPLE_FACTORS - P_2 Treatment:vehicle+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_P_2.mzML; RAW_FILE_NAME=neg_P_2.mzML SUBJECT_SAMPLE_FACTORS - P_3 Treatment:vehicle+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_P_3.mzML; RAW_FILE_NAME=neg_P_3.mzML SUBJECT_SAMPLE_FACTORS - P_4 Treatment:vehicle+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_P_4.mzML; RAW_FILE_NAME=neg_P_4.mzML SUBJECT_SAMPLE_FACTORS - P_5 Treatment:vehicle+cisplatin Genotype=WT; Strain=C57BL/6J; Age=8 weeks; Sex=male; RAW_FILE_NAME=pos_P_5.mzML; RAW_FILE_NAME=neg_P_5.mzML #COLLECTION CO:COLLECTION_SUMMARY The mice were euthanized 72 hours after cisplatin injection, and renal tissues CO:COLLECTION_SUMMARY were collected and frozen at -80℃. CO:SAMPLE_TYPE Kidney CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Wild type C57BL/6J mice were maintained under standard environmental conditions. TR:TREATMENT_SUMMARY A single injection of cisplatin (25 mg/kg, i.p.) was used to induce AKI. To TR:TREATMENT_SUMMARY assess the effect of DMXAA, the mice were injected with DMXAA (10 mg/kg, i.p.) TR:TREATMENT_SUMMARY or vehicle one hour before cisplatin injection, repeated every 24 hours. The TR:TREATMENT_SUMMARY mice were euthanized 72 hours after cisplatin injection, and renal tissues were TR:TREATMENT_SUMMARY collected and frozen at -80℃. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY The samples were taken from the refrigerator at -80℃ and placed on the SP:SAMPLEPREP_SUMMARY prepared dry ice. 30mg samples were taken and 400µL of extract (methanol: SP:SAMPLEPREP_SUMMARY acetonitrile =3:1, pre-cooled at -40℃) was added. After the tissue crushing SP:SAMPLEPREP_SUMMARY instrument was broken, the samples were ultrasonic and stood at 4℃ for 2 SP:SAMPLEPREP_SUMMARY hours. The samples were removed and centrifuged for 15min (12000 rpm, 4℃), SP:SAMPLEPREP_SUMMARY then the same amount was taken and concentrated to dry in vacuum. Then 100µL SP:SAMPLEPREP_SUMMARY 50% methanol aqueous solution (methanol: water =1:1, v/v) was added for SP:SAMPLEPREP_SUMMARY resolution, vortexed for 3min (4℃, 2000rpm), sonicated for 5min, centrifuged SP:SAMPLEPREP_SUMMARY for 15min (12000 rpm, 4℃), and the supernatant was taken for injection SP:SAMPLEPREP_SUMMARY analysis. #CHROMATOGRAPHY CH:INSTRUMENT_NAME Thermo Vanquish liquid chromatograph. CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.7um) CH:CHROMATOGRAPHY_TYPE HILIC #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS Electrospray ionization (ESI) was used to detect positive and negative ions in MS:MS_COMMENTS each sample. The samples were separated by UHPLC and analyzed by mass MS:MS_COMMENTS spectrometry using a Thermo QE HF-X mass spectrometer. Analysis software: MS:MS_COMMENTS Xcalibur version 4.1. MS:ION_MODE POSITIVE MS:MS_RESULTS_FILE ST002390_AN003894_Results.txt UNITS:Average normalized quantitative value Has m/z:Yes Has RT:Yes RT units:Minutes #END