Omics Approaches Towards Discovery of Biomarkers for Early Diagnosis of Tuberculosis Ana Varela Coelho (Presenter) ITQB NOVA Presenter Bio: In 1998, I have started my first research project involving proteomics and changed my research field for mass spectrometry based biological applications, namely on omics approaches. I had coordinated several proteomics projects involving the study of host pathogen interactions. More recently, my research has expanded to the use of other omics approaches involving the discovery of biomarkers for early TB diagnosis and the characterization of Staphylococcus epidermidis infection mechanisms. I have published more than 100 papers in scientific peer-reviewed journals (H index = 27). I am Head of the Proteomics of Non-Model Organisms Lab at ITQB, New University of Lisbon (Portugal). I received my PhD in 1998 from the University of Évora (Portugal) where I was an assistant proprofessor. I have started implementing the MS Facility at ITQB NOVA in 2002, which I coordinated till 2013.

Authors: R Conde1, M Bento1, R Laires1, Anjos S2, C Barroso3, M Villar4, R Macedo5, MJ Simões5, P Lamosa1, PL Fernandes6, Manadas B2, M Matzapetakis1, AV Coelho1
1ITQB NOVA, Oeiras, Portugal; 2CNC.IBILI, University of Coimbra, Coimbra, Portugal, 3CDP Almada-Seixal, ARSLVT, Portugal; 4CDP Venda Nova, ARSLVT, Portugal; 5INSA, Lisboa, Portugal; 6IGC, Oeiras, Portugal

Tuberculosis (TB) is a disease with worldwide presence and a major cause of death in several developing countries. Current diagnostic methodologies often lack specificity and sensitivity, whereas in some cases it takes a long time to obtain a conclusive result. Four metabolites and four proteins were selected as potential biomarkers of TB infection using NMR based metabolomics combined with differential proteomics. These biomarkers are strong candidates for the development of a clinical test.

Introduction

Tuberculosis (TB) is a disease with worldwide presence and a major cause of death in several developing countries. Mycobacterium tuberculosis, an obligate aerobic bacterium with a slow growth at 37°C, is the pathogen responsible for TB. Current diagnostic methodologies often lack specificity and sensitivity, whereas in some cases it takes a long time to obtain a conclusive result. In an effort to develop better diagnostic methods, this work aimed at the discovery of new biomarkers for the diagnosis of TB using a NMR based metabolomics combined with differential proteomics.

Methods

In this study, we acquired 1H NMR and LC-MSMS data of blood serum samples of groups of healthy subjects, individuals with latent TB and of patients with pulmonary and extra-pulmonary TB. Data were obtained using an 800 MHz NMR spectrometer and a TripleTOF 5600 by the SWATH-MS method, respectively.

Results

Results obtained from both omics assays were similar, showing discrimination between controls/latent TB and patients experimental groups. No differences were determined between the two patient groups and the latent tuberculosis subjects. The major metabolic changes reflected in the serum of patients were related with protein biosynthesis, ammonia recycling, amino acids metabolism and glycolysis. By differential proteomics the same three proteins were reported as responsible for the discrimination of controls/latent TB and patients groups. A fourth protein was found diminished only in the extra-pulmonary patients, when compared with the controls/latent TB groups.

Conclusions & Discussion

Some of these results support other studies already published others are novel. They lead to the selection of four metabolites and four proteins as potential biomarkers for further validation.

Acknowledgments: “TBomics - an OMICS approach for diagnosing tuberculosis”, New INDIGO Partnership Programme on Biotechnology applied to Human Health (INDIGO-DBT2-062), ERA-NET Project supported by FCT. To the nurse teams at CDPs Almada-Seixal and Venda Nova.