Summary of Study ST000791

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench,, where it has been assigned Project ID PR000574. The data can be accessed directly via it's Project DOI: 10.21228/M8GD58 This work is supported by NIH grant, U2C- DK119886.


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Study IDST000791
Study TitleIdentifying metabolic adaptations characteristic of multiple myeloma cells via amino acids concentrations from bone marrow plasma
Study SummaryWill be assessing the targeted amino acids concentrations of high risk versus low risk smoldering myeloma patients based on peripheral blood plasma and bone marrow plasma.
Mayo Clinic
Last NameGonsalves
First NameWilson
Address200 First St. SW, Rochester, Minnesota, 55905, USA
Submit Date2017-07-11
Analysis Type DetailLC-MS
Release Date2019-07-17
Release Version1
Wilson Gonsalves Wilson Gonsalves application/zip

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Project ID:PR000574
Project DOI:doi: 10.21228/M8GD58
Project Title:Mayo Pilot and Feasibility: Identifying metabolic adaptations characteristic of multiple myeloma cells via mass spectrometry-based metabolite profiling
Project Summary:Multiple myeloma (MM) is a clonal plasma cell malignancy that remains incurable in most afflicted patients. It can be preceded by an asymptomatic, premalignant stage known as smoldering multiple myeloma (SMM) that does not require therapy but has an increased life-long risk of progression to MM. However, one-third of SMM patients are “high risk” for imminent progression to MM within two years of diagnosis compared to the remainder of SMM patients who continue on an indolent asymptomatic course for several years. The diagnosis of MM cannot be made until they experience overt end-organ damage such as renal failure, lytic bone destruction, anemia and hypercalcemia. Currently, we lack sensitive biomarkers that can identify all SMM patients at high risk of progression to MM. Being able to identify high risk SMM patients could allow us to initiate systemic chemotherapy before they progress to MM. Cancer cells undergo distinct metabolic adaptations to meet the augmented cellular demand for energy and nutrients created by their increased rates of cellular proliferation. The presence of an altered cellular metabolism in clonal PCs from MM patients and its role as an essential factor in the progression of SMM to MM is unknown. We hypothesize that the clonal PCs in high risk SMM patients likely have an altered metabolic phenotype similar to those present in MM patients but different when compared to clonal PCs in the remainder of the SMM patients whose clinical course remains indolent. Thus, two specific aims are proposed in this study: Aim 1 will verify if there are differences in the regulation of the metabolic pathways in clonal PCs from MM patients compared to normal PCs from healthy patients; Aim 2 will assess whether the clonal PCs from high risk SMM patients bear a distinct metabolic phenotype compared to clonal PCs from standard risk SMM patients.
Institute:Mayo Clinic
Last Name:Gonsalves
First Name:Wilson
Address:200 First St. SW, Rochester, Minnesota, 55905, USA


Subject ID:SU000816
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Human


Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

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Collection ID:CO000810
Collection Summary:In order to analyze the metabolites of clonal PCs (intracellular) and BM plasma (extracellular) separately, they have to be separated out from the BM samples. Thus, upon acquiring the BM samples from patients they will be placed in centrifuged at 2500 rpm for 10 minutes to separate out the plasma from the cellular fraction. The separated BM plasma is then stored in separate vials and snap frozen under liquid nitrogen for 20 seconds before storing at -80οC for further analysis. The leftover cellular component present as a pellet will be washed and reconstituted with an equal volume of RPMI 1640 medium. Erythrocytes are lysed using ammonium chloride lysing solution. After incubation on ice for 5 min, the cell suspension is diluted with RPMI medium. The cells are again pelleted by centrifugation and then suspended in RoboSep buffer (250ml PBS, 2% BSA, 1mM EDTA). The clonal CD138 positive PCs are purified using positive selection by mixing the cells with a CD138 positive selection cocktail and anti-CD138 magnetic-activated cell separation microbeads (ROBOSEP™ cell separation system, StemCell Technologies Inc) in the automated RoboSep cell separation system. The purified samples containing only CD138 positive cells are re‐suspended and then centrifuged to form a cell pellet. The goal will be to obtain at least 1-2 x 107 clonal PCs per sample. The cell pellet will be snap frozen under liquid nitrogen for 20 seconds before storing at -80οC for further analysis. Both the BM plasma samples as well as the clonal PCs pellet will be provided to the metabolomics core for sample preparation for LC-MS analysis. For Aim 2, we will be using stored BM samples from SMM patients that have already had their BM plasma and clonal PCs separated from each other and stored at -80οC. It is important to note that all these samples were collected and frozen in a timely manner. Furthermore consistency in sample collection, storage and processing is imperative for the optimal conduct of metabolomics-based experiments. This ensures that all samples being analyzed and compared with each other would have been manipulated similarly, limiting any handling biases. All patient samples in the Predolin Foundation Biobank were collected and stored by following a standardized operating procedure. BM samples were processed for BM plasma separation and clonal PCs enrichment but were then immediately snap frozen for storage at -80οC within approximately 3 hours of collection from the patient. All samples were collected in patients who have been fasting for at least 6 to 8 hours prior. To further ensure consistency in our analysis of this study, we will only use samples that have never been thawed since initial storage to preserve the stability of the metabolites originally present in the samples.
Sample Type:Bone marrow


Treatment ID:TR000830
Treatment Summary:We will use matched BM plasma and purified clonal marrow PCs from the BM samples of SMM patients collected at the time of their diagnosis and stored within the Mayo Clinic Predolin Foundation Biobank. We will select BM samples from patients with SMM who progressed to MM within 2 years of their samples being collected and stored; this will constitute the high risk SMM group. We will also select BM samples from SMM patients who have not progressed to MM within at least 2 years of follow up of their samples being collected and stored; this will constitute the standard risk SMM group. The strength of this approach is that we will utilize stored SMM samples from one of the most comprehensively characterized monoclonal gammopathy biobanks available. Secondly, all samples will be obtained from collection dates at least 2 years prior in order to ensure adequate follow-up time to assess their current clinical status. This would avoid the need to prospectively collect samples from SMM patients and clinically follow them for a number of years before we are able to gauge who were clinically high or standard risk for progression. There are over 700 SMM patients who have had their BM samples collected and stored in the biobank from 1996 to 2013; more than half these patients have progressed to MM.

Sample Preparation:

Sampleprep ID:SP000823
Sampleprep Summary:amino acids concentrations

Combined analysis:

Analysis ID AN001259
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity
Column Waters Acquity BEH C18 (150 x 2.1mm,1.7um)
MS instrument type Triple quadrupole
MS instrument name Thermo Quantum Ultra
Units uM


Chromatography ID:CH000877
Instrument Name:Waters Acquity
Column Name:Waters Acquity BEH C18 (150 x 2.1mm,1.7um)
Chromatography Type:Reversed phase


MS ID:MS001152
Analysis ID:AN001259
Instrument Name:Thermo Quantum Ultra
Instrument Type:Triple quadrupole