Summary of Study ST002876
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 PR001798. The data can be accessed directly via it's Project DOI: 10.21228/M89D97 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 | ST002876 |
| Study Title | High Level Expression of NSD2 Creates a Metabolic Dependency in Multiple Myeloma |
| Study Summary | Multiple myeloma (MM) is a malignancy of plasma cells with several molecular subtypes and variable prognosis. Despite therapeutic advances, most patients ultimately relapse due to drug resistance. Frontline treatments for MM target malignant cells based on their differentiated B cell nature, but not the underlying genetic lesions. Chromosomal translocation t(4;14), observed in 15% of MM patients, results in overexpression of the histone methyltransferase NSD2, which contributes to MM pathogenesis by promoting an oncogenic transcriptional program and is associated with a worse prognosis. A genome-wide CRISPR based functional screen in isogenic MM cell lines with high and low NSD2 expression identified the mitochondrial enzyme adenylate kinase 2 (AK2) as a NSD2-driven MM cell dependency. AK2 loss in t(4;14) MM cells induced apoptosis and inhibited cell growth in vitro and in vivo. Consistent with a defect in shuttling ATP from the mitochondria to intracellular utilization sites, AK2 depletion impaired ATP-dependent protein folding in the ER and increased MM cell sensitivity to the proteasome inhibitor bortezomib. Furthermore, AK2 suppression decreased intracellular NAD(H) phosphorylation resulting in lower NADP(H) levels. Cytosolic NADPH is necessary for reducing thioredoxin, an essential cofactor for ribonucleotide reductase which is critical for deoxyribonucleotides (dNTP) synthesis. Consequently, AK2 deficiency in MM cells resulted in dNTP pool depletion and induced DNA replication stress. Creatine phosphorylation by mitochondrial creatine kinase is an alternative route for shuttling ATP from the mitochondria to the cytosol. Metabolomics analysis revealed decreased levels of creatine and accumulation of its precursor guanadoacetate in NSD2 high cells, in association with elevated levels of S-adenosyl homocysteine (SAH) indicating consumption of the carbon donor S-Adenosyl methionine (SAM). This along with the 6-fold increase in genome wide H3K36me2 levels and 40% increase in DNA methylation levels in NSD2 high cells suggested that overexpression of NSD2 redirected one-carbon metabolism to the epigenome and away from the SAM-dependent creatine synthesis. Therefore, decreased creatine levels in NSD2 overexpressing cells underlie the increased reliance on AK2. Correspondingly, supplementation with exogenous creatine restored NADP(H) levels and rescued AK2 deficient cells from apoptosis. These findings revealed a novel metabolic susceptibility in t(4;14) MM and provided insight into a novel therapeutic strategy to improve patient prognosis. We performed metabolomic analysis of multiple myeloma cell lines with either high or low NSD2 levels with or without NSD2 knockdown. |
| Institute | University of Florida |
| Last Name | Licht |
| First Name | Jonathan |
| Address | 2033 MOWRY RD STE 145 GAINESVILLE FL 32610 |
| jdlicht@ufl.edu | |
| Phone | (352) 273-8143 |
| Submit Date | 2023-09-07 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-10-30 |
| Release Version | 1 |
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Sample Preparation:
| Sampleprep ID: | SP002995 |
| Sampleprep Summary: | Soluble metabolites were extracted directly from snap-frozen cell pellets using cold acetonitrile/water (80/20, vol/vol) at a dilution of 10,000 cells/ul solvent. Samples were vortexed and incubated at -80C overnight to precipitate proteins and subsequently thawed, vortexed, and centrifuged at 18,000 x g for 30 min at 4C to pellet debris. The supernatants were transferred to new vials and analyzed by high-performance liquid chromatography (HPLC) and high-resolution mass spectrometry and tandem mass spectrometry (MS/MS). |
