Summary of Study ST000548
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 PR000402. The data can be accessed directly via it's Project DOI: 10.21228/M83890 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST000548 |
| Study Title | Replication study: The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarate |
| Study Summary | The Reproducibility Project: Cancer Biology seeks to address growing concerns about reproducibility in scientific research by conducting replications of selected experiments from a number of high-profile papers in the field of cancer biology. The papers, which were published between 2010 and 2012, were selected on the basis of citations and Altmetric scores (Errington et al., 2014). This Registered Report describes the proposed replication plan of key experiments from “The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarate” by Ward and colleagues, published in Cancer Cell in 2010 (Ward et al., 2010). The experiments that will be replicated are those reported in Figures 2, 3 and 5. Ward and colleagues demonstrate the mutations in isocitrate dehydrogenase 2 (IDH2), commonly found in acute myeloid leukemia (AML), abrogate the enzyme’s wild-type activity and confer to the mutant neomorphic activity that produces the oncometabolite 2-hydroxyglutarate (2-HG) (Figures 2 and 3). They then show that elevated levels of 2-HG are correlated with mutations in IDH1 and IDH2in AML patient samples (Figure 5). The Reproducibility Project: Cancer Biology is a collaboration between the Center for Open Science and Science Exchange and the results of the replications will be published by eLife. |
| Institute | University of California, Davis |
| Department | Genome and Biomedical Sciences Facility |
| Laboratory | WCMC Metabolomics Core |
| Last Name | Fiehn |
| First Name | Oliver |
| Address | Health Sciences Drive, Davis, California, 95616, USA |
| ofiehn@ucdavis.edu | |
| Phone | (530) 754-8258 |
| Submit Date | 2017-01-06 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | d |
| Analysis Type Detail | GC-MS |
| Release Date | 2017-07-10 |
| Release Version | 1 |
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Treatment:
| Treatment ID: | TR000584 |
| Treatment Summary: | 239T cells (ATCC CRL-3216) were grown in DMEM (Invitrogen) with 10% FBS (Hyclone) at -37°C in 10% CO2 and transfected with pcDNA3.1, pcDNA3.1-IDH2WT, pcDNA3.1-IDH2R172K (Invitrogen and Origene) with Lipofectamine 2000 (Invitrogen) according to manufacturer instructions. 6x10^5 cells were seeded in 6 well plates for protocol 1 and 3.5x10^6 cells were seeded in 10 cm plates for protocol 2. Identity of all vectors was confirmed by sequencing and vector integrity with agarose gel electrophoresis.The common feature of IDH1 and IDH2 mutations.pdf |
| Treatment Protocol Filename: | 239T cells (ATCC CRL-3216) were grown in DMEM (Invitrogen) with 10% FBS (Hyclone) at 37°C in 10% CO2 and transfected with pcDNA3.1, pcDNA3.1-IDH2WT, pcDNA3.1-IDH2R172K (Invitrogen and Origene) with Lipofectamine 2000 (Invitrogen) according to manufacturer instructions. |
| Cell Harvesting: | 24-48 hours after transfection |
