Partial Least Squares Discriminant Analysis of NMR spectra acquired on blood serum samples.

<p>Scores (<b>A</b>) and weights (on LV1, <b>B</b>, and LV2, <b>C</b>) plots obtained from OSC-PLS-DA performed on the NMR spectra of 44 blood serum samples. Group A (solid red, 19 samples): patients at diagnosis; group C1 (solid blue, 10 samples): sustained remission and group C2 (empty blue, 15 samples): in relapse after chemotherapy. Cho: choline; Cre: creatinine; Pyr: pyruvate; Ala: alanine; 2HiB: 2-hydroxyisobutyrate; Lac: lactate; 2HB: 2-hydroxybutyrate. H-xan: hypoxantine; Glu: glutamate; Gln: glutamine; Ace: acetate.</p

Partial Least Squares Discriminant Analysis of NMR spectra acquired on blood serum samples.

<p>Scores (<b>A</b> and <b>B</b>) and weights (on LV1; <b>C</b> and <b>D</b>) plots obtained from OSC-PLS-DA performed on the NMR spectra of 37 and 42 blood serum samples for the comparison of groups B versus C1 (<b>A</b> and <b>C</b>) and B versus C2 (<b>B</b> and <b>D</b>). Group B (solid green, 27 samples): patients after chemotherapy; group C1 (solid blue, 10 samples): sustained remission; group C2 (empty blue, 15 samples): in relapse after chemotherapy. Cho: choline; Suc: succinate; Pyr: pyruvate; Ace: acetate; 2HiB: 2-hydroxyisobutyrate; Car: carnitine; AcCar: acetylcarnitine.</p

Patients characteristics.

<p>Footnotes <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056422#pone-0056422-t001" target="_blank">table 1</a>. Serum reference ranges:- beta2 microglobulin 0.5–4.0 mg/l; serum creatinine 45–110 µmol/l (0.5 to 1.2 mg/dl). Paraprotein types detected by immunofixation of serum and urine: GLO IgG lambda, no flc in urine; GKU IgG kappa, flc in urine; NS non secretory, no Ig detected in blood or urine; KUO kappa flc only detected in urine but not serum; AKU IgA kappa, kappa flc in urine; ALO IgA lambda, no flc in urine; KUS kappa flc only detected in serum and urine; ALU IgA lambda, flc in urine; DLU IgD lambda, flc in urine.</p

Proton NMR spectra and Partial Least Squares Discriminant Analysis of blood serum samples.

<p>Representative sections of proton NMR spectra at the diagnosis (red), remission (green) and prolonged remission (blue) of one multiple myeloma patient. (<b>A</b>) up-field region (0.75–4.25 ppm), (<b>B</b>) down-field region (5.1–8.9 ppm), 20 times increased intensity compared to (A). (<b>C</b>) Scores plot obtained from OSC-PLS-DA performed on the NMR spectra of 71 blood serum samples. Group A (solid red, 19 samples): patients at diagnosis; group B (solid green, 27 samples): patients after chemotherapy; group C1 (solid blue, 10 samples): sustained remission and group C2 (empty blue, 15 samples): in relapse after chemotherapy.</p

Partial Least Squares Discriminant Analysis of NMR spectra acquired on blood serum samples.

<p>Scores (<b>A</b>) and weights (on LV1; <b>B</b>) plots obtained from OSC-PLS-DA performed on the NMR spectra of 46 blood serum samples. Group A (solid red, 19 samples): patients at diagnosis; Group B (solid green, 27 samples): patients after chemotherapy. Lac: lactate; Cho: choline; Cre: creatinine; Suc: succinate; Ace: acetate; Ala: alanine; 3HB: 3-hydroxybutyrate; 2HB: 2-hydroxybutyrate.</p

Blood serum concentration of acetylcarnitine and carnitine.

<p>Blood serum concentration of acetylcarnitine (AcCAR) and carnitine (CAR) in MM patients at diagnosis, in remission and after relapse of active disease following chemotherapy. Data shown as mean ± s.e.m. Statistical significance calculated according to Kruskal-Wallis one-way ANOVA (p<0.05).</p

Proton NMR-Based Metabolite Analyses of Archived Serial Paired Serum and Urine Samples from Myeloma Patients at Different Stages of Disease Activity Identifies Acetylcarnitine as a Novel Marker of Active Disease

<div><p>Background</p><p>Biomarker identification is becoming increasingly important for the development of personalized or stratified therapies. Metabolomics yields biomarkers indicative of phenotype that can be used to characterize transitions between health and disease, disease progression and therapeutic responses. The desire to reproducibly detect ever greater numbers of metabolites at ever diminishing levels has naturally nurtured advances in best practice for sample procurement, storage and analysis. Reciprocally, since many of the available extensive clinical archives were established prior to the metabolomics era and were not processed in such an ‘ideal’ fashion, considerable scepticism has arisen as to their value for metabolomic analysis. Here we have challenged that paradigm.</p> <p>Methods</p><p>We performed proton nuclear magnetic resonance spectroscopy-based metabolomics on blood serum and urine samples from 32 patients representative of a total cohort of 1970 multiple myeloma patients entered into the United Kingdom Medical Research Council Myeloma IX trial.</p> <p>Findings</p><p>Using serial paired blood and urine samples we detected metabolite profiles that associated with diagnosis, post-treatment remission and disease progression. These studies identified carnitine and acetylcarnitine as novel potential biomarkers of active disease both at diagnosis and relapse and as a mediator of disease associated pathologies.</p> <p>Conclusions</p><p>These findings show that samples conventionally processed and archived can provide useful metabolomic information that has important implications for understanding the biology of myeloma, discovering new therapies and identifying biomarkers potentially useful in deciding the choice and application of therapy.</p> </div

Partial Least Squares Discriminant Analysis of NMR spectra acquired on blood serum samples.

<p>Scores (<b>A</b>) and weights (on LV1; <b>B</b>) plots obtained from OSC-PLS-DA performed on the NMR spectra of 52 blood serum samples. Group B (solid green, 27 samples): patients after chemotherapy; group C1 (solid blue, 10 samples): sustained remission; group C2 (empty blue, 15 samples): in relapse after chemotherapy. Lac: lactate; Gly: glycine; Tau: taurine; Cho: choline; Suc: succinate; Pyr: pyruvate; Ace: acetate; 2HiB: 2-hydroxyisobutyrate.</p

SALMON: Solvent Accessibility, Ligand binding, and Mapping of ligand Orientation by NMR Spectroscopy

Quinone oxidoreductase 2 (NQO2) binds the prodrug tretazicar (also known as CB1954, 5-(aziridin-1-yl)-2,4-dinitrobenzamide), which exhibits a profound antitumor effect in human cancers when administered together with caricotamide. X-ray structure determination allowed for two possible orientations of the ligand. Here we describe a new NMR method, SALMON (solvent accessibility, ligand binding, and mapping of ligand orientation by NMR spectroscopy), based on waterLOGSY to determine the orientation of a ligand bound to a protein by mapping its solvent accessibility, which was used to unambiguously determine the orientation of CB1954 in NQO2

Hypoxia-Like Signatures Induced by BCR-ABL Potentially Alter the Glutamine Uptake for Maintaining Oxidative Phosphorylation

<div><p>The Warburg effect is probably the most prominent metabolic feature of cancer cells, although little is known about the underlying mechanisms and consequences. Here, we set out to study these features in detail in a number of leukemia backgrounds. The transcriptomes of human CB CD34⁺ cells transduced with various oncogenes, including BCR-ABL, MLL-AF9, FLT3-ITD, NUP98-HOXA9, STAT5A and KRAS^G12V were analyzed in detail. Our data indicate that in particular BCR-ABL, KRAS^G12V and STAT5 could impose hypoxic signaling under normoxic conditions. This coincided with an upregulation of glucose importers SLC2A1/3, hexokinases and HIF1 and 2. NMR-based metabolic profiling was performed in CB CD34⁺ cells transduced with BCR-ABL versus controls, both cultured under normoxia and hypoxia. Lactate and pyruvate levels were increased in BCR-ABL-expressing cells even under normoxia, coinciding with enhanced glutaminolysis which occurred in an HIF1/2-dependent manner. Expression of the glutamine importer SLC1A5 was increased in BCR-ABL⁺ cells, coinciding with an increased susceptibility to the glutaminase inhibitor BPTES. Oxygen consumption rates also decreased upon BPTES treatment, indicating a glutamine dependency for oxidative phosphorylation. The current study suggests that BCR-ABL-positive cancer cells make use of enhanced glutamine metabolism to maintain TCA cell cycle activity in glycolytic cells.</p></div