Ultrahigh-Resolution
NMR with Water Signal Suppression
for a Deeper Understanding of the Action of Antimetabolic Drugs on
Diffuse Large B‑Cell Lymphoma
posted on 2022-02-04, 20:11authored byGildas Bertho, Leonardo Lordello, Xi Chen, Covadonga Lucas-Torres, Imed Eddine Oumezziane, Cédric Caradeuc, Mathieu Baudin, Stéphanie Nuan-Aliman, Catherine Thieblemont, Véronique Baud, Nicolas Giraud
Ultrahigh-resolution
NMR has recently attracted considerable attention
in the field of complex samples analysis. Indeed, the implementation
of broadband homonuclear decoupling techniques has allowed us to greatly
simplify crowded 1H spectra, yielding singlets for almost
every proton site from the analyzed molecules. Pure shift methods
have notably shown to be particularly suitable for deciphering mixtures
of metabolites in biological samples. Here, we have successfully implemented
a new pure shift pulse sequence based on the PSYCHE method, which
incorporates a block for solvent suppression that is suitable for
metabolomics analysis. The resulting experiment allows us to record
ultrahigh-resolution 1D NOESY 1H spectra of biofluids with
suppression of the water signal, which is a crucial step for highlighting
metabolite mixtures in an aqueous phase. We have successfully recorded
pure shift spectra on extracellular media of diffuse large B-cell
lymphoma (DLBCL) cells. Despite a lower sensitivity, the resolution
of pure shift data was found to be better than that of the standard
approach, which provides a more detailed vision of the exo-metabolome.
The statistical analyses carried out on the resulting metabolic profiles
allow us to successfully highlight several metabolic pathways affected
by these drugs. Notably, we show that Kidrolase plays a major role
in the metabolic pathways of this DLBCL cell line.