{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002119","ANALYSIS_ID":"AN003469","VERSION":"1","CREATED_ON":"March 31, 2022, 8:01 am"},

"PROJECT":{"PROJECT_TITLE":"MNK2 deficiency potentiates beta-cell regeneration via hypertranslation","PROJECT_SUMMARY":"Regenerating pancreatic beta-cells is a potential curative approach for diabetes. We previously identified the small molecule CID661578 as a potent inducer of beta-cell regeneration but its target and mechanism of action have remained unknown. We now screened 257 million yeast clones and determined that CID661578 targets MAP kinase-interacting serine/threonine kinase 2 (MNK2), an interaction we genetically validated in vivo. CID661578 increased beta-cell neogenesis from ductal cells in zebrafish, neonatal pig islet aggregates and human pancreatic ductal organoids. Mechanistically, we found that CID661578 boosts protein synthesis and regeneration by blocking MNK2 from binding eIF4G in the translation initiation complex at the mRNA cap. Unexpectedly, this blocking activity augmented eIF4E-phosphorylation depending on MNK1 and bolstered the interaction between eIF4E and eIF4G, which is necessary for both hypertranslation and beta-cell regeneration. Taken together, our findings demonstrate a targetable role of MNK2-controlled translation in beta-cell regeneration, a role that warrants further investigation in diabetes.","INSTITUTE":"North Carolina State University","LAST_NAME":"Liu","FIRST_NAME":"Xiaojing","ADDRESS":"Polk Hall, RM 128","EMAIL":"xliu68@ncsu.edu","PHONE":"9195154387"},

"STUDY":{"STUDY_TITLE":"Metabolomics analysis of zebrafish response to CID661578 treatment","STUDY_SUMMARY":"zebrafish larvae were treated with DMSO or CID661578 for 24 hours prior to global metabolomics analysis (n=6). Metabolites were extracted from pools of 10 zebrafish larvae at 5 dpf using a 80% methanol-based extraction method. Samples were dried in speed vac and stored in -80C freezer until ready for LC-MS analysis.","INSTITUTE":"North Carolina State University","LAST_NAME":"Liu","FIRST_NAME":"Xiaojing","ADDRESS":"Polk Hall, RM 128","EMAIL":"xliu68@ncsu.edu","PHONE":"9195154387"},

"SUBJECT":{"SUBJECT_TYPE":"Fish","SUBJECT_SPECIES":"Danio rerio","TAXONOMY_ID":"7955"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"DMSO1",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO1.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO2",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO2.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO3",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO3.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO4",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO4.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO5",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO5.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO6",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO6.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID1",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID1.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID2",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID2.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID3",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID3.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID4",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID4.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID5",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID5.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID6",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID6.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO1_Pos",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO1_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO2_Pos",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO2_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO3_Pos",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO3_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO4_Pos",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO4_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO5_Pos",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO5_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO6_Pos",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO6_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID1_Pos",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID1_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID2_Pos",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID2_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID3_Pos",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID3_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID4_Pos",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID4_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID5_Pos",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID5_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID6_Pos",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID6_Pos.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO1_Neg",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO1_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO2_Neg",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO2_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO3_Neg",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO3_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO4_Neg",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO4_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO5_Neg",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO5_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"DMSO6_Neg",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"RAW_FILE_NAME":"DMSO6_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID1_Neg",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID1_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID2_Neg",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID2_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID3_Neg",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID3_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID4_Neg",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID4_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID5_Neg",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID5_Neg.raw"}
},
{
"Subject ID":"-",
"Sample ID":"CID6_Neg",
"Factors":{"Treatment":"CID661578"},
"Additional sample data":{"RAW_FILE_NAME":"CID6_Neg.raw"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"zebrafish larvae were treated with DMSO or CID661578 for 24 hours prior to global metabolomics analysis (n=6). Metabolites were extracted from pools of 10 zebrafish larvae at 5 dpf using a 80% methanol/water-based extraction method. Detailed metabolite extraction was described previously using 80% methanol/water as solvent (PMID: 24410464, PMID: 24894601, PMID: 25795660) and dry pellets were stored in -80 °C freezer until ready for LC-MS analysis. Pellets were reconstituted into 30 μL of sample solvent (water:methanol:acetonitrile, 2:1:1, v/v), and 5 μL was injected into the LC-MS.","SAMPLE_TYPE":"new"},

"TREATMENT":{"TREATMENT_SUMMARY":"Chemicals (DMSO orCID661578) were added to the E3 medium for larvae to reach final concentration of 10 uM. Pools of 10 wild-type (WT) larvae at 5 dpf were used for each of the 6 independent biological replicates for DMSO or CID661578 treatment from 4-5 dpf."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Intracellular metabolites were harvested as described previously using 80% methanol/water as solvent (PMID: 24410464, PMID: 24894601, PMID: 25795660) and dry pellets were stored in -80 °C freezer until ready for LC-MS analysis. For metabolite analysis, pellets were reconstituted into 30 μL of sample solvent (water:methanol:acetonitrile, 2:1:1, v/v), and 5 μL was injected into the LC-MS."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"HILIC method is for general metabolomics analysis.","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Waters Xbridge amide (100 × 2.1 mm i.d., 3.5 μm)"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Plus Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"It was operated in the full-scan mode (positive ion mode only) with the resolution set at 70 000 (at m/z 200).LC-MS data were analyzed using Sieve 2.0 (Thermo Scientific), and the integrated area under metabolite peak was used to compare the relative abundance of each metabolite in different samples in the same batch."},

"MS_METABOLITE_DATA":{
"Units":"ion counts",

"Data":[{"Metabolite":"acetyl-CoA","DMSO1_Pos":"133063.821","DMSO2_Pos":"125855.106","DMSO3_Pos":"79763.07","DMSO4_Pos":"68044.05","DMSO5_Pos":"96745.416","DMSO6_Pos":"78967.134","CID1_Pos":"88387.719","CID2_Pos":"79334.541","CID3_Pos":"135358.299","CID4_Pos":"91848.972","CID5_Pos":"96118.131","CID6_Pos":"71600.565"},{"Metabolite":"Propanoyl-CoA","DMSO1_Pos":"31221.399","DMSO2_Pos":"43384.341","DMSO3_Pos":"48726.195","DMSO4_Pos":"13642.125","DMSO5_Pos":"23289.864","DMSO6_Pos":"20121.597","CID1_Pos":"55769.403","CID2_Pos":"50613.171","CID3_Pos":"90764.583","CID4_Pos":"196322.619","CID5_Pos":"58947.492","CID6_Pos":"42712.101"},{"Metabolite":"crotonoyl-CoA","DMSO1_Pos":"0","DMSO2_Pos":"0","DMSO3_Pos":"0","DMSO4_Pos":"0","DMSO5_Pos":"0","DMSO6_Pos":"0","CID1_Pos":"0","CID2_Pos":"0","CID3_Pos":"0","CID4_Pos":"0","CID5_Pos":"0","CID6_Pos":"0"},{"Metabolite":"isobutyryl-CoA","DMSO1_Pos":"26807.508","DMSO2_Pos":"40590.513","DMSO3_Pos":"48781.239","DMSO4_Pos":"17954.34","DMSO5_Pos":"35660.157","DMSO6_Pos":"50901.837","CID1_Pos":"109079.532","CID2_Pos":"62423.67","CID3_Pos":"138773.781","CID4_Pos":"91589.316","CID5_Pos":"149320.842","CID6_Pos":"114692.787"},{"Metabolite":"Isovaleryl-CoA","DMSO1_Pos":"28256.169","DMSO2_Pos":"48897.501","DMSO3_Pos":"58418.079","DMSO4_Pos":"25756.686","DMSO5_Pos":"49561.011","DMSO6_Pos":"52287.744","CID1_Pos":"148009.854","CID2_Pos":"107519.577","CID3_Pos":"184149.873","CID4_Pos":"204500.922","CID5_Pos":"235056.744","CID6_Pos":"203805.444"},{"Metabolite":"malonyl-CoA","DMSO1_Pos":"5929.911","DMSO2_Pos":"5647.824","DMSO3_Pos":"6359.547","DMSO4_Pos":"10227.117","DMSO5_Pos":"12042.84","DMSO6_Pos":"1005.96","CID1_Pos":"11133.408","CID2_Pos":"13926.138","CID3_Pos":"8915.946","CID4_Pos":"2133.798","CID5_Pos":"6277.836","CID6_Pos":"6003.504"},{"Metabolite":"(S)-3-hydroxybutanoyl-CoA","DMSO1_Pos":"0","DMSO2_Pos":"0","DMSO3_Pos":"0","DMSO4_Pos":"0","DMSO5_Pos":"0","DMSO6_Pos":"0","CID1_Pos":"0","CID2_Pos":"0","CID3_Pos":"0","CID4_Pos":"0","CID5_Pos":"0","CID6_Pos":"0"},{"Metabolite":"Succinyl-CoA","DMSO1_Pos":"42573.408","DMSO2_Pos":"55697.07","DMSO3_Pos":"57417.471","DMSO4_Pos":"53847.744","DMSO5_Pos":"51236.475","DMSO6_Pos":"48101.928","CID1_Pos":"72384.771","CID2_Pos":"62165.325","CID3_Pos":"89462.058","CID4_Pos":"138883.08","CID5_Pos":"63569.85","CID6_Pos":"51982.62"},{"Metabolite":"Hydroxymethylglutaryl-CoA","DMSO1_Pos":"0","DMSO2_Pos":"0","DMSO3_Pos":"0","DMSO4_Pos":"0","DMSO5_Pos":"0","DMSO6_Pos":"0","CID1_Pos":"0","CID2_Pos":"0","CID3_Pos":"0","CID4_Pos":"0","CID5_Pos":"0","CID6_Pos":"0"}],

"Metabolites":[{"Metabolite":"acetyl-CoA","Chemical formula":"C23H38N7O17P3S"},{"Metabolite":"Propanoyl-CoA","Chemical formula":"C24H40N7O17P3S"},{"Metabolite":"crotonoyl-CoA","Chemical formula":"C25H40N7O17P3S"},{"Metabolite":"isobutyryl-CoA","Chemical formula":"C25H42N7O17P3S"},{"Metabolite":"Isovaleryl-CoA","Chemical formula":"C26H44N7O17P3S"},{"Metabolite":"malonyl-CoA","Chemical formula":"C24H38N7O19P3S"},{"Metabolite":"(S)-3-hydroxybutanoyl-CoA","Chemical formula":"C25H42N7O18P3S"},{"Metabolite":"Succinyl-CoA","Chemical formula":"C25H40N7O19P3S"},{"Metabolite":"Hydroxymethylglutaryl-CoA","Chemical formula":"C27H44N7O20P3S"}]
}

}