Novel protocol for metabolomics data normalization and biomarker discovery in human tears
Por:
Serrano-Marín J, Marín-del Barrio S, Iglesias A, Lillo J, Garrigós C, Capó T, Reyes-Resina I, Alkozi HA, Cascante M, Sánchez-Navés J, Franco R and Bernal-Casas D
Publicada:
28 jul 2025
Ahead of Print:
1 mar 2025
Resumen:
Objectives Human tear analysis holds promise for biomarker discovery, but its clinical utility is hindered by the lack of standardized reference values, limiting interindividual comparisons. This study aimed at developing a protocol for normalizing metabolomic data from human tears, enhancing its potential for biomarker identification.Methods Tear metabolomic profiling was conducted on 103 donors (64 females, 39 males, aged 18-82 years) without ocular pathology, using the AbsoluteIDQ (TM) p180 Kit for targeted metabolomics. A predictive normalization model incorporating age, sex, and fasting time was developed to correct for interindividual variability. Key metabolites from six compound families (amino acids, biogenic amines, acylcarnitines, lysophosphatidylcholines, phosphatidylcholines, and sphingomyelins) were identified as normalization references. The approach was validated using Linear Discriminant Analysis (LDA) to test its ability to classify donor sex based on metabolite concentrations.Results Metabolite concentrations exhibited significant interindividual variability. The normalization model, which predicted metabolite concentrations based on a reference "concomitant" metabolite from each compound family, successfully reduced this variability. Using the ratio of observed-to-predicted concentrations, the model enabled robust comparisons across individuals. LDA classification of donor sex using acylcarnitine C4 achieved 78 % accuracy, correctly identifying 92 % of female donors. This approach outperformed traditional statistical and machine learning methods (Lasso logistic regression and Random Forest classification) in sex discrimination based on tear metabolomics.Conclusions This novel normalization protocol significantly improves the reliability of tear metabolomics by enabling standardized interindividual comparisons. The approach facilitates biomarker discovery by mitigating variability in metabolite concentrations and may be extended to other biological fluids, enhancing its applicability in precision medicine.
Filiaciones:
Serrano-Marín J:
Molecular Neurobiology Laboratory, Universitat de Barcelona, Barcelona, Spain
Marín-del Barrio S:
Department of Biochemistry and Molecular Biomedicine, Faculty of Biology, Universitat de Barcelona (UB), Barcelona, Spain
Institute of Biomedicine of University of Barcelona (IBUB), University of Barcelona (UB), Barcelona, Spain
CIBEREHD, Network Center for Hepatic and Digestive Diseases, Spanish National Health Institute Carlos III (ISCIII), Madrid, Spain
Iglesias A:
Molecular Neurobiology Laboratory, Universitat de Barcelona, Barcelona, Spain
Lillo J:
Molecular Neurobiology Laboratory, Universitat de Barcelona, Barcelona, Spain
CiberNed, Network Center for Neurodegenerative Diseases, Spanish National Health Institute Carlos III (ISCIII), Madrid, Spain
Garrigós C:
Molecular Neurobiology Laboratory, Universitat de Barcelona, Barcelona, Spain
Capó T:
Molecular Neurobiology Laboratory, Universitat de Barcelona, Barcelona, Spain
Reyes-Resina I:
CiberNed, Network Center for Neurodegenerative Diseases, Spanish National Health Institute Carlos III (ISCIII), Madrid, Spain
Department of Biochemistry and Physiology, Faculty of Pharmacy and Food Sciences, Universitat de Barcelona (UB), Barcelona, Spain
Alkozi HA:
Department of Optometry, College of Applied Medical Sciences, Qassim University, Almulida, Qassim, Saudi Arabia
Cascante M:
Department of Biochemistry and Molecular Biomedicine, Faculty of Biology, Universitat de Barcelona (UB), Barcelona, Spain
Institute of Biomedicine of University of Barcelona (IBUB), University of Barcelona (UB), Barcelona, Spain
CIBEREHD, Network Center for Hepatic and Digestive Diseases, Spanish National Health Institute Carlos III (ISCIII), Madrid, Spain
Sánchez-Navés J:
Department of Ophthalmology, Oftalmedic and I.P.O. Institute of Ophthalmology, Palma de Mallorca, Spain
Franco R:
Department of Biochemistry and Molecular Biomedicine, Faculty of Biology, Universitat de Barcelona (UB), Barcelona, Spain
CiberNed, Network Center for Neurodegenerative Diseases, Spanish National Health Institute Carlos III (ISCIII), Madrid, Spain
School of Chemistry, Universitat de Barcelona, Barcelona, Spain
Bernal-Casas D:
Independent Researcher, Barcelona, Spain
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