NanoLC-MS/MS analyses of urinary desmosine, hydroxylysylpyridinoline and lysylpyridinoline as biomarkers for chronic graft-versus-host disease.
Chronic graft-versus-host disease (cGVHD) is a common and potentially lethal complication of allogeneic hematopoietic stem cell transplantation (HSCT). cGVHD as well as the transplant procedure itself (chemotherapy with or without radiotherapy) can lead to the degradation of connective tissue components such as elastin and collagen. The catabolism of these structural proteins releases desmosine (DES), lysylpyridinoline (LP), hydroxylysylpyridonoline (HP), and related pyridinium-based cross-linkers analogues that could represent potential biomarkers for cGVHD. This study reports the development of a sensitive liquid chromatography/tandem mass spectrometry method for the simultaneous analysis of N-propyl derivatives of DES, HP, and LP. The concentrations of free and total forms of urinary DES, HP, and LP were determined using synthetic deuterated internal standards. This method enabled accurate quantitation of these pyridinium-based cross-linkers from as little as 100 microL of urine with detection limits of 0.03-0.10 ng/mL. These compounds were analyzed in urine samples from three groups of patients: (1) Healthy volunteers, (2) Autologous HSCT recipients (who cannot develop cGVHD), and (3) Allogeneic HSCT recipients at onset of cGHVD. These analyses revealed that the urinary concentrations of DES, HP, and LP in the autologous recipients were greater or equal to the cGVHD group although both groups showed marked increase in the levels of these compounds compared to healthy individuals. These results suggest that the chemotherapy treatment has significant effects on the turnover of elastin and collagen, and that these biomarkers could be effective during prospective analyses to determine the onset of cGVHD.
Institute for Research in Immunology and Cancer, Université de Montreal, P.O. Box 6128, Station Centre-ville, Canada H3C 3J7.
Michel
M
Boutin
Imran
I
Ahmad
Marjo
M
Jauhiainen
Nathalie
N
Lachapelle
Claude
C
Rondeau
Jean
J
Roy
Pierre
P
Thibault
D000328:Adult; D000368:Aged; D000596:Amino Acids / Q000652:urine*; D015415:Biomarkers / Q000652:urine; D002853:Chromatography, Liquid / Q000379:methods*; D003895:Desmosine / Q000652:urine*; D005260:Female; D006086:Graft vs Host Disease / Q000652:urine*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006801:Humans; D008297:Male; D008875:Middle Aged; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000596:Amino Acids; D015415:Biomarkers; D003895:Desmosine; C015484:pyridinoline; C036020:deoxypyridinoline