{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002474","ANALYSIS_ID":"AN004041","VERSION":"1","CREATED_ON":"February 10, 2023, 9:58 pm"},

"PROJECT":{"PROJECT_TITLE":"Retinol Dehydrogenase 10 Reduction Mediated Retinol Metabolism Disorder Promotes Diabetic Cardiomyopathy in Male Mice.","PROJECT_SUMMARY":"In this study, we identify disordered cardiac retinol metabolism in type 2 diabetic male mice and patients characterized by retinol overload, all-trans retinoic acid deficiency. By supplementing type 2 diabetic male mice with retinol or all-trans retinoic acid, we demonstrate that both cardiac retinol overload and all-trans retinoic acid deficiency promote diabetic cardiomyopathy. Mechanistically, by constructing cardiomyocyte-specific conditional retinol dehydrogenase 10-knockout male mice and overexpressing retinol dehydrogenase 10 in male type 2 diabetic mice via adeno-associated virus, we verify that the reduction in cardiac retinol dehydrogenase 10 is the initiating factor for cardiac retinol metabolism disorder and results in diabetic cardiomyopathy. Therefore, we suggest that the reduction of cardiac retinol dehydrogenase 10 and its mediated disorder of cardiac retinol metabolism is a new mechanism underlying diabetic cardiomyopathy.","INSTITUTE":"Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University","LAST_NAME":"Yandi","FIRST_NAME":"Wu","ADDRESS":"74, Zhongshan second street","EMAIL":"wuyd3@mail2.sysu.edu.cn","PHONE":"15622158754","PUBLICATIONS":"Retinol Dehydrogenase 10 Reduction Mediated Retinol Metabolism Disorder Promotes Diabetic Cardiomyopathy in Male Mice.","CONTRIBUTORS":"Yandi Wu; Tongsheng Huang; Xinghui Li; Conghui Shen; Honglin Ren; Haiping Wang; Teng Wu; Xinlu Fu; Shijie Deng; Ziqi Feng; Shijie Xiong; Hui Li; Saifei Gao; Zhenyu Yang; Fei Gao; Lele Dong; Jianding Cheng & Weibin Cai"},

"STUDY":{"STUDY_TITLE":"Retinol Dehydrogenase 10 Reduction Mediated Retinol Metabolism Disorder Promotes Diabetic Cardiomyopathy in Male Mice.","STUDY_SUMMARY":"In this study, we identify disordered cardiac retinol metabolism in type 2 diabetic male mice and patients characterized by retinol overload, all-trans retinoic acid deficiency. By supplementing type 2 diabetic male mice with retinol or all-trans retinoic acid, we demonstrate that both cardiac retinol overload and all-trans retinoic acid deficiency promote diabetic cardiomyopathy. Mechanistically, by constructing cardiomyocyte-specific conditional retinol dehydrogenase 10-knockout male mice and overexpressing retinol dehydrogenase 10 in male type 2 diabetic mice via adeno-associated virus, we verify that the reduction in cardiac retinol dehydrogenase 10 is the initiating factor for cardiac retinol metabolism disorder and results in diabetic cardiomyopathy. Therefore, we suggest that the reduction of cardiac retinol dehydrogenase 10 and its mediated disorder of cardiac retinol metabolism is a new mechanism underlying diabetic cardiomyopathy.","INSTITUTE":"Department of Biochemistry, Zhongshan School of Medicine, Sun Yat-sen University","LAST_NAME":"Yandi","FIRST_NAME":"Wu","ADDRESS":"74, Zhongshan second street","EMAIL":"wuyd3@mail2.sysu.edu.cn","PHONE":"15622158754","PUBLICATIONS":"Retinol Dehydrogenase 10 Reduction Mediated Retinol Metabolism Disorder Promotes Diabetic Cardiomyopathy in Male Mice."},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"db/m; db/db; db/db+retinol; db/db+retinoic acid; db/db+AAV9-RDH10; RDH10-cKO; RDH10fl/fl","AGE_OR_AGE_RANGE":"32-week; 36-week; 21-week","GENDER":"Male"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"32-week db/m-1",
"Factors":{"Genotype":"db/m+","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-1"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/m-2",
"Factors":{"Genotype":"db/m+","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-2"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/m-3",
"Factors":{"Genotype":"db/m+","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-3"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/db-1",
"Factors":{"Genotype":"db/db","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-4"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/db-2",
"Factors":{"Genotype":"db/db","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-5"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/db-3",
"Factors":{"Genotype":"db/db","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-6"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/db-4",
"Factors":{"Genotype":"db/db","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-7"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/db-5",
"Factors":{"Genotype":"db/db","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-8"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/db-6",
"Factors":{"Genotype":"db/db","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-9"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/db+Rol-1",
"Factors":{"Genotype":"db/db","Treatment":"retinol"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-10"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/db+Rol-2",
"Factors":{"Genotype":"db/db","Treatment":"retinol"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-11"}
},
{
"Subject ID":"-",
"Sample ID":"32-week db/db+Rol-3",
"Factors":{"Genotype":"db/db","Treatment":"retinol"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-12"}
},
{
"Subject ID":"-",
"Sample ID":"RDH10-FL-1",
"Factors":{"Genotype":"RDH10fl/fl","Treatment":"tamoxifen"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-13"}
},
{
"Subject ID":"-",
"Sample ID":"RDH10-FL-2",
"Factors":{"Genotype":"RDH10fl/fl","Treatment":"tamoxifen"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-14"}
},
{
"Subject ID":"-",
"Sample ID":"RDH10-FL-3",
"Factors":{"Genotype":"RDH10fl/fl","Treatment":"tamoxifen"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-15"}
},
{
"Subject ID":"-",
"Sample ID":"RDH10-CKO-1",
"Factors":{"Genotype":"RDH10fl/fl, MYH6-iCre","Treatment":"tamoxifen"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-16"}
},
{
"Subject ID":"-",
"Sample ID":"RDH10-CKO-2",
"Factors":{"Genotype":"RDH10fl/fl, MYH6-iCre","Treatment":"tamoxifen"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-17"}
},
{
"Subject ID":"-",
"Sample ID":"RDH10-CKO-3",
"Factors":{"Genotype":"RDH10fl/fl, MYH6-iCre","Treatment":"tamoxifen"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-18"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/m-1",
"Factors":{"Genotype":"db/m+","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-19"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/m-2",
"Factors":{"Genotype":"db/m+","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-20"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/m-3",
"Factors":{"Genotype":"db/m+","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-21"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/db-1",
"Factors":{"Genotype":"db/db","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-22"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/db-2",
"Factors":{"Genotype":"db/db","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-23"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/db-3",
"Factors":{"Genotype":"db/db","Treatment":"no"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-24"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/db+atRA-1",
"Factors":{"Genotype":"db/db","Treatment":"all-trans retinoic acid"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-25"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/db+atRA-2",
"Factors":{"Genotype":"db/db","Treatment":"all-trans retinoic acid"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-26"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/db+atRA-3",
"Factors":{"Genotype":"db/db","Treatment":"all-trans retinoic acid"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-27"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/db+aav9-RDH10-1",
"Factors":{"Genotype":"db/db","Treatment":"aav9-rdh10"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-28"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/db+aav9-RDH10-2",
"Factors":{"Genotype":"db/db","Treatment":"aav9-rdh11"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-29"}
},
{
"Subject ID":"-",
"Sample ID":"36-week db/db+aav9-RDH10-3",
"Factors":{"Genotype":"db/db","Treatment":"aav9-rdh12"},
"Additional sample data":{"RAW_FILE_NAME":"20201203-Retinoic acid-Sample-30"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Mice were fasted for 12 hours before sampling and testing. All animal experiments were approved by the Animal Care and Ethics Committee of Zhongshan School of Medicine, Sun Yat-sen University, and followed the National Institutes of Health Guidelines on the Care and Use of Animals (the protocol number is SYSU-IACUC-2019-B027)","SAMPLE_TYPE":"Cardiac tissue"},

"TREATMENT":{"TREATMENT_SUMMARY":"RDH10-cKO mice: RDH10-cKO mice, which contain both RDH10fl/fl and MYH6-iCre, were bred by RDH10fl/fl mice and MYH6-iCre mice and were injected tamoxifen intraperitoneally (50 mg/kg, T2859, Sigma -Aldrich, St. Louis, MO) for 5 consecutive days from 5 weeks of age. Age-matched male RDH10fl/fl mice that also received TMX injections served as normal controls for RDH10-cKO mice. Animal treatments: Mice in the Rol treatment group received Rol gavage (800 IU/each, 17772, Sigma-Aldrich, St. Louis, MO) every two days from 8 weeks of age. Mice in the atRA treatment group received atRA intraperitoneal injection (5 mg/kg body weight, R2625, Sigma-Aldrich, St. Louis, MO) daily from 8 weeks of age. Gene therapy: A recombinant AAV9 vector carrying the mouse RDH10 sequence (AAV9-RDH10, DZ-AAV-Rdh10-OE, Dongze, Hanbio Inc, Shanghai, China) was used to overexpress RDH10. 0.8*10^11 vg/per animal of AA9-RDH10 was transferred into T2DM mice, respectively, by tail vein injection at the age of 16 weeks."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Sample preparation Homogenized the heart tissue in 200μL of cold NaCl solution (0.9%) for 5 seconds, add 100μL retinol-D4 (IR-23012, IsoSciences, Ambler, PA, USA) as internal standard for Rol and atRA, and retinyl acetate (46958, Sigma-Aldrich, St. Louis, MO) as internal standard for retinyl esters, then homogenize for another 5 seconds. For Rol and at RA, 1mL formic acid n-hexane solution (1%) was added to the homogenizing to facilitate phase separation by a 5-minutes vortex and 5-min centrifugation (16,200 g). The supernatants were collected in a new tube and blown dry with nitrogen followed by re-dilution with 100μL 70% methanol. For retinyl esters, 24 μL 0.5 M NaOH solution was added to tissue homogenate, followed by 1 mL n-hexane. The mixture was votexed 5 min and centrifuged at 16,200 g for 5 min. The supernatants were collected in a new tube and blown dry with nitrogen followed by re-dilution with 100μL 100% methanol. The processes above should be protected from the light."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Sciex Triple Quad TM 4500MD MS","COLUMN_NAME":"Phenomenex Kinetex C18 (50 x 2.1mm,1.7um)","SOLVENT_A":"40% acetonitrile/30% methanol/30% water; 0.1 % formic acid","SOLVENT_B":"55% acetonitrile/30% methanol/15% water; 0.1 % formic acid","FLOW_GRADIENT":"initial conditions 75% B; from 0 to 3.5min linear increase to 95% B; between 3.5 and 4.5min 95% B was retained; at 4.6 min back to initial conditions with 75% B; finally, 75% B was held from 4.6 to 5.9min","FLOW_RATE":"0.2ml/min","COLUMN_TEMPERATURE":"25"},

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

"MS":{"INSTRUMENT_NAME":"ABI Sciex 5500 QTrap","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"The MS conditions were as follows: electrospray ionization (ESI) under positive mode; nebulizer gas: nitrogen; curtain gas,30 psi; ion spray voltage, 4000V; temperature, 400ºC; gas 1 and gas 2, 35 and 40 psi, respectively; collision gas, 10 psi. The parameters of the mass spectrometer were optimized, and the multiple reaction monitoring (MRM) transitions of retinol, all-trans-retinoic acid, as well as D4-retinol were chosen as 269.2>93.1, 273.1>94.0, and 301.2>123.1, respectively. The MRM transitions for retinyl esters were chosen as 329.3>269.3 for retinyl acetate, 524.4>268.1 for retinyl palmitate (16:0), 552.5>268.2 for retinyl stearate (18:0), 522.4>268, retinyl palmitoleate (16:1), and retinyl oleate (18:1). The quantification was performed using the calibration curve."},

"MS_METABOLITE_DATA":{
"Units":"ng/ml",

"Data":[{"Metabolite":"all-trans retinoic acid","32-week db/m-1":"0.4225","32-week db/m-2":"0.2801","32-week db/m-3":"0.465","32-week db/db-1":"0.3109","32-week db/db-2":"0.2123","32-week db/db-3":"0.2443","32-week db/db-4":"0.2457","32-week db/db-5":"0.2077","32-week db/db-6":"0.2331","32-week db/db+Rol-1":"0.8069","32-week db/db+Rol-2":"0.1854","32-week db/db+Rol-3":"0.1568","RDH10-FL-1":"0.3221","RDH10-FL-2":"0.2427","RDH10-FL-3":"0.738","RDH10-CKO-1":"0.1387","RDH10-CKO-2":"0.1652","RDH10-CKO-3":"0.2088","36-week db/m-1":"0.2639","36-week db/m-2":"0.2968","36-week db/m-3":"0.4983","36-week db/db-1":"0.1145","36-week db/db-2":"0.2287","36-week db/db-3":"0.1264","36-week db/db+atRA-1":"0.4869","36-week db/db+atRA-2":"0.3228","36-week db/db+atRA-3":"0.3495","36-week db/db+aav9-RDH10-1":"0.2571","36-week db/db+aav9-RDH10-2":"0.2941","36-week db/db+aav9-RDH10-3":"0.2698"},{"Metabolite":"retinol","32-week db/m-1":"2.7287","32-week db/m-2":"3.9078","32-week db/m-3":"8.4067","32-week db/db-1":"11.4845","32-week db/db-2":"4.3457","32-week db/db-3":"11.8574","32-week db/db-4":"8.2282","32-week db/db-5":"7.8723","32-week db/db-6":"7.5237","32-week db/db+Rol-1":"1.7974","32-week db/db+Rol-2":"8.3747","32-week db/db+Rol-3":"13.6601","RDH10-FL-1":"5.056","RDH10-FL-2":"1.3794","RDH10-FL-3":"8.2627","RDH10-CKO-1":"4.3456","RDH10-CKO-2":"6.1914","RDH10-CKO-3":"9.9269","36-week db/m-1":"5.4485","36-week db/m-2":"7.9932","36-week db/m-3":"8.1485","36-week db/db-1":"6.4338","36-week db/db-2":"12.4819","36-week db/db-3":"7.311","36-week db/db+atRA-1":"5.9866","36-week db/db+atRA-2":"4.9836","36-week db/db+atRA-3":"6.6919","36-week db/db+aav9-RDH10-1":"7.2647","36-week db/db+aav9-RDH10-2":"7.4051","36-week db/db+aav9-RDH10-3":"9.6483"}],

"Metabolites":[{"Metabolite":"all-trans retinoic acid","retention index":"None","quantified m/z":"301.2>123.1","PubChem ID":"444795","KEGG ID":"C00777"},{"Metabolite":"retinol","retention index":"None","quantified m/z":"269.2>93.1","PubChem ID":"445354","KEGG ID":"C00473"}]
}

}