Journal
JOURNAL OF PROTEOME RESEARCH
Volume 16, Issue 4, Pages 1669-1682Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.6b01038
Keywords
BALf (bronchoalveolar lavage fluid); BOS (brOnchiolitis obliterans syndrome); CLAD (chronic lung allograft dysfunction); NMR-based metabolomics
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Funding
- Fondazione CARIPLO (Milan, Italy) [2013-0820]
- Fondazione CARIPLO [2015-0763]
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This report describes: the application of NMR spectroscopy to the profiling of metabolites in bronchoalveolar lavage fluid (BALf) of lung transplant recipients without bronchiolitis obliterans syndrome (BOS) (stable, n 10), and with BOS at different degrees of severity (BOS Op) n = 10; BOS I, n = 10). Through the fine;tuning of a number of parameters concerning both sample preparation/processing and variations of spectra acquisition modes, an efficient and reproducible protocol was designed for the screening of metabolites in a pulmonary fluid that should reflect the status of airway inflammation/injury. Exploiting the combination of mono- and bidimensional NMR experiments, 38 polar metabolites, including amino acids; Krebs cycle intermediates, mono- and disaccharides, nucleotides, and phovholipid precursors, were unequivocally identified. To, determine which signature could be correlated with the onset of BOS, the metabolites' content of the above recipients was analyzed by multivariate (PCA and OPLSDA) statistical methods. PCA analysis (almost) totally differentiated S from BOS I, and this discrimination was significantly improved by the application of OPLS-DA, whose model was characterized by excellent fit and prediction values (R-2 = 0.99 and Q(2) = 0.88). The analysis of S vs BOS Op and of BOS Op vs BOS I samples showed a clear discrimination of considered cohorts, although with a poorer efficiency compared to those measured for S vs BOS I patients. The data shown in this work assess the suitability of the NMR approach in monitoring different pathological Tung conditions.
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