Summary of Study ST002474
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001597. The data can be accessed directly via it's Project DOI: 10.21228/M88H9Z This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST002474 |
| 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 | Zhongshan School of Medicine, Sun Yat-sen University |
| Department | Dept. of Biochemistry |
| Last Name | Yandi |
| First Name | Wu |
| Address | 74, Zhongshan second street |
| wuyd3@mail2.sysu.edu.cn | |
| Phone | 15622158754 |
| Submit Date | 2023-02-08 |
| Publications | Retinol Dehydrogenase 10 Reduction Mediated Retinol Metabolism Disorder Promotes Diabetic Cardiomyopathy in Male Mice. |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-02-13 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001597 |
| Project DOI: | doi: 10.21228/M88H9Z |
| 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: | 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 |
Subject:
| Subject ID: | SU002564 |
| 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 |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype | Treatment |
|---|---|---|---|
| SA247790 | 36-week db/db+aav9-RDH10-1 | db/db | aav9-rdh10 |
| SA247791 | 36-week db/db+aav9-RDH10-2 | db/db | aav9-rdh11 |
| SA247792 | 36-week db/db+aav9-RDH10-3 | db/db | aav9-rdh12 |
| SA247793 | 36-week db/db+atRA-3 | db/db | all-trans retinoic acid |
| SA247794 | 36-week db/db+atRA-1 | db/db | all-trans retinoic acid |
| SA247795 | 36-week db/db+atRA-2 | db/db | all-trans retinoic acid |
| SA247796 | 36-week db/db-2 | db/db | no |
| SA247797 | 36-week db/db-3 | db/db | no |
| SA247798 | 32-week db/db-2 | db/db | no |
| SA247799 | 36-week db/db-1 | db/db | no |
| SA247800 | 32-week db/db-1 | db/db | no |
| SA247801 | 32-week db/db-6 | db/db | no |
| SA247802 | 32-week db/db-5 | db/db | no |
| SA247803 | 32-week db/db-4 | db/db | no |
| SA247804 | 32-week db/db-3 | db/db | no |
| SA247805 | 32-week db/db+Rol-1 | db/db | retinol |
| SA247806 | 32-week db/db+Rol-2 | db/db | retinol |
| SA247807 | 32-week db/db+Rol-3 | db/db | retinol |
| SA247808 | 32-week db/m-2 | db/m+ | no |
| SA247809 | 32-week db/m-3 | db/m+ | no |
| SA247810 | 32-week db/m-1 | db/m+ | no |
| SA247811 | 36-week db/m-1 | db/m+ | no |
| SA247812 | 36-week db/m-2 | db/m+ | no |
| SA247813 | 36-week db/m-3 | db/m+ | no |
| SA247784 | RDH10-CKO-3 | "RDH10fl/fl, MYH6-iCre" | tamoxifen |
| SA247785 | RDH10-CKO-2 | "RDH10fl/fl, MYH6-iCre" | tamoxifen |
| SA247786 | RDH10-CKO-1 | "RDH10fl/fl, MYH6-iCre" | tamoxifen |
| SA247787 | RDH10-FL-2 | RDH10fl/fl | tamoxifen |
| SA247788 | RDH10-FL-1 | RDH10fl/fl | tamoxifen |
| SA247789 | RDH10-FL-3 | RDH10fl/fl | tamoxifen |
| Showing results 1 to 30 of 30 |
Collection:
| Collection ID: | CO002557 |
| 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 ID: | TR002576 |
| 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. |
Sample Preparation:
| Sampleprep ID: | SP002570 |
| 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. |
Combined analysis:
| Analysis ID | AN004041 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Sciex Triple Quad TM 4500MD MS |
| Column | Phenomenex Kinetex C18 (50 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | QTRAP |
| MS instrument name | ABI Sciex 5500 QTrap |
| Ion Mode | POSITIVE |
| Units | ng/ml |
Chromatography:
| Chromatography ID: | CH002989 |
| Instrument Name: | Sciex Triple Quad TM 4500MD MS |
| Column Name: | Phenomenex Kinetex C18 (50 x 2.1mm,1.7um) |
| Column Temperature: | 25 |
| 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 |
| Solvent A: | 40% acetonitrile/30% methanol/30% water; 0.1 % formic acid |
| Solvent B: | 55% acetonitrile/30% methanol/15% water; 0.1 % formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003788 |
| Analysis ID: | AN004041 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | QTRAP |
| MS Type: | ESI |
| 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. |
| Ion Mode: | POSITIVE |
