#METABOLOMICS WORKBENCH zhanglsh8_20221001_195924 DATATRACK_ID:3484 STUDY_ID:ST002302 ANALYSIS_ID:AN003759 PROJECT_ID:PR001475 VERSION 1 CREATED_ON October 3, 2022, 9:13 am #PROJECT PR:PROJECT_TITLE Integrated metabolomics and lipidomics study of patients with atopic dermatitis PR:PROJECT_TITLE in response to dupilumab PR:PROJECT_SUMMARY Background: Atopic dermatitis (AD) is one of the most common chronic PR:PROJECT_SUMMARY inflammatory skin diseases. Dupilumab, a monoclonal antibody that targets the PR:PROJECT_SUMMARY interleukin (IL)-4 and IL-13 receptors, has been widely used in AD because of PR:PROJECT_SUMMARY its efficacy. However, metabolic changes occurring in patients with AD in PR:PROJECT_SUMMARY response to dupilumab remains unknown. In this study, we integrated metabolomics PR:PROJECT_SUMMARY and lipidomics analyses with clinical data to explore potential metabolic PR:PROJECT_SUMMARY alterations associated with dupilumab therapeutic efficacy. In addition, we PR:PROJECT_SUMMARY investigate whether the development of treatment side effects was linked to the PR:PROJECT_SUMMARY dysregulation of metabolic pathways. Methods: A total of 33 patients with AD PR:PROJECT_SUMMARY were included in the current study, with serum samples collected before and PR:PROJECT_SUMMARY after treatment with dupilumab. Comprehensive metabolomic and lipidomic analyses PR:PROJECT_SUMMARY have previously been developed to identify serum metabolites (including lipids) PR:PROJECT_SUMMARY that vary among treatment groups. An orthogonal partial least squares PR:PROJECT_SUMMARY discriminant analysis model was established to screen for differential PR:PROJECT_SUMMARY metabolites and metabolites with variable importance in projection > 1 and p < PR:PROJECT_SUMMARY 0.05 were considered potential metabolic biomarkers. MetaboAnalyst 5.0 was used PR:PROJECT_SUMMARY to identify related metabolic pathways. Patients were further classified into PR:PROJECT_SUMMARY two groups, well responders (n = 19) and poor responders (n = 14), to identify PR:PROJECT_SUMMARY differential metabolites between the two groups. Results: The results revealed PR:PROJECT_SUMMARY significant changes in serum metabolites before and after 16 weeks of dupilumab PR:PROJECT_SUMMARY treatment. Variations in the metabolic profile were more significant in the PR:PROJECT_SUMMARY well-responder group than in the poor-responder group. Pathway enrichment PR:PROJECT_SUMMARY analysis revealed that differential metabolites derived from the well-responder PR:PROJECT_SUMMARY group were mainly involved in glycerophospholipid metabolism, valine, leucine PR:PROJECT_SUMMARY and isoleucine biosynthesis, the citrate cycle, arachidonic acid metabolism, PR:PROJECT_SUMMARY pyrimidine metabolism, and sphingolipid metabolism. Conclusion: Serum metabolic PR:PROJECT_SUMMARY profiles of patients with AD varied significantly after treatment with PR:PROJECT_SUMMARY dupilumab. Differential metabolites and their related metabolic pathways may PR:PROJECT_SUMMARY provide clues for understanding the effects of dupilumab on patient metabolism. PR:INSTITUTE Peking Union Medical College Hospital, Chinese Academy of Medical Sciences PR:LAST_NAME Zhang PR:FIRST_NAME Lishan PR:ADDRESS No.1 Shuaifuyuan Wangfujing Dongcheng District, Beijing, 100730, China. PR:EMAIL 429647356@qq.com PR:PHONE +86-18612636397 #STUDY ST:STUDY_TITLE Integrated metabolomics and lipidomics study of patients with atopic dermatitis ST:STUDY_TITLE in response to dupilumab ST:STUDY_SUMMARY Background: Atopic dermatitis (AD) is one of the most common chronic ST:STUDY_SUMMARY inflammatory skin diseases. Dupilumab, a monoclonal antibody that targets the ST:STUDY_SUMMARY interleukin (IL)-4 and IL-13 receptors, has been widely used in AD because of ST:STUDY_SUMMARY its efficacy. However, metabolic changes occurring in patients with AD in ST:STUDY_SUMMARY response to dupilumab remains unknown. In this study, we integrated metabolomics ST:STUDY_SUMMARY and lipidomics analyses with clinical data to explore potential metabolic ST:STUDY_SUMMARY alterations associated with dupilumab therapeutic efficacy. In addition, we ST:STUDY_SUMMARY investigate whether the development of treatment side effects was linked to the ST:STUDY_SUMMARY dysregulation of metabolic pathways. Methods: A total of 33 patients with AD ST:STUDY_SUMMARY were included in the current study, with serum samples collected before and ST:STUDY_SUMMARY after treatment with dupilumab. Comprehensive metabolomic and lipidomic analyses ST:STUDY_SUMMARY have previously been developed to identify serum metabolites (including lipids) ST:STUDY_SUMMARY that vary among treatment groups. An orthogonal partial least squares ST:STUDY_SUMMARY discriminant analysis model was established to screen for differential ST:STUDY_SUMMARY metabolites and metabolites with variable importance in projection > 1 and p < ST:STUDY_SUMMARY 0.05 were considered potential metabolic biomarkers. MetaboAnalyst 5.0 was used ST:STUDY_SUMMARY to identify related metabolic pathways. Patients were further classified into ST:STUDY_SUMMARY two groups, well responders (n = 19) and poor responders (n = 14), to identify ST:STUDY_SUMMARY differential metabolites between the two groups. Results: The results revealed ST:STUDY_SUMMARY significant changes in serum metabolites before and after 16 weeks of dupilumab ST:STUDY_SUMMARY treatment. Variations in the metabolic profile were more significant in the ST:STUDY_SUMMARY well-responder group than in the poor-responder group. Pathway enrichment ST:STUDY_SUMMARY analysis revealed that differential metabolites derived from the well-responder ST:STUDY_SUMMARY group were mainly involved in glycerophospholipid metabolism, valine, leucine ST:STUDY_SUMMARY and isoleucine biosynthesis, the citrate cycle, arachidonic acid metabolism, ST:STUDY_SUMMARY pyrimidine metabolism, and sphingolipid metabolism. Conclusion: Serum metabolic ST:STUDY_SUMMARY profiles of patients with AD varied significantly after treatment with ST:STUDY_SUMMARY dupilumab. Differential metabolites and their related metabolic pathways may ST:STUDY_SUMMARY provide clues for understanding the effects of dupilumab on patient metabolism. ST:INSTITUTE Peking Union Medical College Hospital, Chinese Academy of Medical Sciences ST:LAST_NAME Zhang ST:FIRST_NAME Lishan ST:ADDRESS No.1 Shuaifuyuan Wangfujing Dongcheng District, Beijing, 100730, China. ST:EMAIL 429647356@qq.com ST:PHONE +86-18612636397 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:AGE_OR_AGE_RANGE >=18 SU:GENDER Male and female SU:HUMAN_RACE Chinese SU:HUMAN_ETHNICITY Han SU:HUMAN_TRIAL_TYPE observational study SU:HUMAN_MEDICATIONS Dupilumab SU:HUMAN_INCLUSION_CRITERIA 1.Age ≥ 18 years of age 2.Dermatologist diagnosis of moderate to severe AD, SU:HUMAN_INCLUSION_CRITERIA EASI≥16 at baseline 3.Eligible to receive dupilumab therapy for AD in SU:HUMAN_INCLUSION_CRITERIA accordance with the guidelines. Patients who are eligible were treated with a SU:HUMAN_INCLUSION_CRITERIA fixed schedule of 300mg dupilumab in 2-week intervals. Patients who did not SU:HUMAN_INCLUSION_CRITERIA achieve 16-week therapy were excluded. 4.During the whole treatment process, the SU:HUMAN_INCLUSION_CRITERIA requirements for diet and exercise are roughly the same as before treatment, so SU:HUMAN_INCLUSION_CRITERIA as to keep the body healthy and balanced 5.A 30-day washout period of systemic SU:HUMAN_INCLUSION_CRITERIA medications preceded treatment SU:HUMAN_EXCLUSION_CRITERIA 1Evidence of other skin diseases except for AD at baseline 2.Pregnancy or breast SU:HUMAN_EXCLUSION_CRITERIA feeding, 3.Patients with permanent severe diseases, especially those affecting SU:HUMAN_EXCLUSION_CRITERIA the immune system, except asthma 4.Patients with severe mental illness SU:HUMAN_EXCLUSION_CRITERIA 5.Evidence of chronic metabolic disease, including Obesity, diabetes, fatty SU:HUMAN_EXCLUSION_CRITERIA liver, osteoporosis, atherosclerotic cardiovascular and cerebrovascular SU:HUMAN_EXCLUSION_CRITERIA diseases, and metabolic-related cancers (breast, colorectal, pancreatic, colon, SU:HUMAN_EXCLUSION_CRITERIA and prostate cancer). 6.Application of other systemic medications during SU:HUMAN_EXCLUSION_CRITERIA treatment #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 - QC01 Treatment:Control RAW_FILE_NAME=QC01.mzXML SUBJECT_SAMPLE_FACTORS - QC02 Treatment:Control RAW_FILE_NAME=QC02.mzXML SUBJECT_SAMPLE_FACTORS - QC03 Treatment:Control RAW_FILE_NAME=QC03.mzXML SUBJECT_SAMPLE_FACTORS - QC04 Treatment:Control RAW_FILE_NAME=QC04.mzXML SUBJECT_SAMPLE_FACTORS - QC05 Treatment:Control RAW_FILE_NAME=QC05.mzXML SUBJECT_SAMPLE_FACTORS - QC06 Treatment:Control RAW_FILE_NAME=QC06.mzXML SUBJECT_SAMPLE_FACTORS - QC07 Treatment:Control RAW_FILE_NAME=QC07.mzXML SUBJECT_SAMPLE_FACTORS - A1 Treatment:Before dupilumab treatment RAW_FILE_NAME=A1.mzXML SUBJECT_SAMPLE_FACTORS - A2 Treatment:Before dupilumab treatment RAW_FILE_NAME=A2.mzXML SUBJECT_SAMPLE_FACTORS - A3 Treatment:Before dupilumab treatment RAW_FILE_NAME=A3.mzXML SUBJECT_SAMPLE_FACTORS - A4 Treatment:Before dupilumab treatment RAW_FILE_NAME=A4.mzXML SUBJECT_SAMPLE_FACTORS - A5 Treatment:Before dupilumab treatment RAW_FILE_NAME=A5.mzXML SUBJECT_SAMPLE_FACTORS - A6 Treatment:Before dupilumab treatment RAW_FILE_NAME=A6.mzXML SUBJECT_SAMPLE_FACTORS - 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B10 Treatment:After dupilumab treatment RAW_FILE_NAME=B10.mzXML SUBJECT_SAMPLE_FACTORS - B11 Treatment:After dupilumab treatment RAW_FILE_NAME=B11.mzXML SUBJECT_SAMPLE_FACTORS - B12 Treatment:After dupilumab treatment RAW_FILE_NAME=B12.mzXML SUBJECT_SAMPLE_FACTORS - B13 Treatment:After dupilumab treatment RAW_FILE_NAME=B13.mzXML SUBJECT_SAMPLE_FACTORS - B14 Treatment:After dupilumab treatment RAW_FILE_NAME=B14.mzXML SUBJECT_SAMPLE_FACTORS - B15 Treatment:After dupilumab treatment RAW_FILE_NAME=B15.mzXML SUBJECT_SAMPLE_FACTORS - B16 Treatment:After dupilumab treatment RAW_FILE_NAME=B16.mzXML SUBJECT_SAMPLE_FACTORS - B17 Treatment:After dupilumab treatment RAW_FILE_NAME=B17.mzXML SUBJECT_SAMPLE_FACTORS - B18 Treatment:After dupilumab treatment RAW_FILE_NAME=B18.mzXML SUBJECT_SAMPLE_FACTORS - B19 Treatment:After dupilumab treatment RAW_FILE_NAME=B19.mzXML SUBJECT_SAMPLE_FACTORS - B20 Treatment:After dupilumab treatment RAW_FILE_NAME=B20.mzXML SUBJECT_SAMPLE_FACTORS - B21 Treatment:After dupilumab treatment RAW_FILE_NAME=B21.mzXML SUBJECT_SAMPLE_FACTORS - B22 Treatment:After dupilumab treatment RAW_FILE_NAME=B22.mzXML SUBJECT_SAMPLE_FACTORS - B23 Treatment:After dupilumab treatment RAW_FILE_NAME=B23.mzXML SUBJECT_SAMPLE_FACTORS - B24 Treatment:After dupilumab treatment RAW_FILE_NAME=B24.mzXML SUBJECT_SAMPLE_FACTORS - B25 Treatment:After dupilumab treatment RAW_FILE_NAME=B25.mzXML SUBJECT_SAMPLE_FACTORS - B26 Treatment:After dupilumab treatment RAW_FILE_NAME=B26.mzXML SUBJECT_SAMPLE_FACTORS - B27 Treatment:After dupilumab treatment RAW_FILE_NAME=B27.mzXML SUBJECT_SAMPLE_FACTORS - B28 Treatment:After dupilumab treatment RAW_FILE_NAME=B28.mzXML SUBJECT_SAMPLE_FACTORS - B29 Treatment:After dupilumab treatment RAW_FILE_NAME=B29.mzXML SUBJECT_SAMPLE_FACTORS - B30 Treatment:After dupilumab treatment RAW_FILE_NAME=B30.mzXML SUBJECT_SAMPLE_FACTORS - B31 Treatment:After dupilumab treatment RAW_FILE_NAME=B31.mzXML SUBJECT_SAMPLE_FACTORS - B32 Treatment:After dupilumab treatment RAW_FILE_NAME=B32.mzXML SUBJECT_SAMPLE_FACTORS - B33 Treatment:After dupilumab treatment RAW_FILE_NAME=B33.mzXML #COLLECTION CO:COLLECTION_SUMMARY We recruited 33 patients diagnosed with moderate to severe atopic dermatitis at CO:COLLECTION_SUMMARY the dermatology outpatient clinic of the Peking Union Medical College Hospital CO:COLLECTION_SUMMARY from March 2021 to February 2022. Serum samples from each participant were CO:COLLECTION_SUMMARY obtained after overnight fasting at the outpatient clinic of the Peking Union CO:COLLECTION_SUMMARY Medical College Hospital and were collected before and after 16 weeks of CO:COLLECTION_SUMMARY dupilumab treatment. The serum samples were immediately frozen at -80°C until CO:COLLECTION_SUMMARY analysis. CO:SAMPLE_TYPE Blood (serum) #TREATMENT TR:TREATMENT_SUMMARY All the enrolled participants were treated with dupilumab for 16 weeks. TR:TREATMENT Biologics Formulation TR:TREATMENT_ROUTE in 2-week intervals TR:TREATMENT_DOSE 300mg dupilumab TR:TREATMENT_DOSEVOLUME 300mg dupilumab #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 100 μL of sample was transferred to an EP tube. After the addition of 400 μL SP:SAMPLEPREP_SUMMARY of extract solution (acetonitrile: methanol = 1: 1, containing SP:SAMPLEPREP_SUMMARY isotopically-labelled internal standard mixture), the samples were vortexed for SP:SAMPLEPREP_SUMMARY 30 s, sonicated for 10 min in ice-water bath, and incubated for 1 h at -40 ℃ SP:SAMPLEPREP_SUMMARY to precipitate proteins. Then the sample was centrifuged at 12000 SP:SAMPLEPREP_SUMMARY rpm(RCF=13800(×g),R= 8.6cm) for 15 min at 4 ℃. The resulting supernatant was SP:SAMPLEPREP_SUMMARY transferred to a fresh glass vial for analysis. The quality control (QC) sample SP:SAMPLEPREP_SUMMARY was prepared by mixing an equal aliquot of the supernatants from all of the SP:SAMPLEPREP_SUMMARY samples. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Waters Acquity BEH C8 (100 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive HF-X Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS - MS:MS_RESULTS_FILE ST002302_AN003759_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes #END