Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Lung-2015 Baltimore, USA July 13 - 15, 2015 Du Toit Loots Tuberculosis: adaptations of man and microbe in order to outcompete and survive History of TB • Robert Koch - 1882 History of TB 150 million years ago Jurassic period • 3 million years ago – infection in early hominids (TB meningitis in Homo erectus) • 1.7 million years ago Migration out of East Africa Evolutionary timeline = TB timeline Tuberculosis: Shocking Statistics • Infectious disease of primarily lungs – M. tuberculosis • 2nd most deadliest infectious disease (after HIV) • ⅓ of global population infected • 9 million new cases per year • 1,4 million deaths per year of which 25% are HIV coinfected • MDR-TB, XDR-TB is on the rise in many 3rd world countries • Despite fervent research efforts since it's discovery in 1882 – TB is still considered a global epidemic • New approaches are needed = Metabolomics? Metabolomics • Unbiased, identification and quantification of ALL intra- and extra-cellular metabolites (small molecule intermediates and products of metabolism) present in a biological system/sample (cell, body fluid, tissue, organism), using highly selective and sensitive analytical methods (such as liquid/gas chromatography and mass spectrometry), in conjunction with bioinformatics for identifying new metabolite markers, at a specific point in time and under certain conditions. • Uses: Identifying compounds / markers of a perturbation Systems Biology DNA RNA Proteins Metabolites Systems / Integrative Biology NWU-Human Metabolomics GC-MS: • Agilent GC-MSD • Leco Pegasus 4D GCxGC-TOFMS • Leco GC-TOFMS CE: • Agilent CE NMR: • Bruker 500Mhz LC-MS: • Agilent 6410 LC-QQQ • Agilent 6460 LC-QQQ • Agilent 6220 LC-TOF • Agilent 6510 LC-QTOF • Agilent LC-MSD XCT Plus IonTrap • Agilent LC-UV(DAD) • Synapt G2 SI UPLC QTOF Plus Ion Mobility Untargeted and Semi-targeted metabolomics Untargeted: • GCxGC-TOFMS • UPLC-TOFMS Ion Mobility • NMR Semi-Targeted GC-MS/LC-MS: • Free Fatty Acids • Long Chain Fatty Acids • Organic Acids • Acylcarnitines • Amino Acids Samples • Blood • Urine • Tissue • Sputum • Cell Cultures • Plant • Skin Secretions (Frogs) • Etc… Metabolomics workflow Metabolite Markers Our Journey 1) TB Characterization: J.C. Schoeman & Du T. Loots. (2011). Improved disease characterization and diagnostics using metabolomics: A review. J Cell Tissue Research, 11(1): 2673-2683. 2) TB Diagnostics: I. Olivier & Du T. Loots. (2011). An overview of tuberculosis treatments and diagnostics. What role could metabolomics play? J Cell Tissue Research, 11(1): 2655-2671. 1) Total Lipid Extraction Method 2) Total Metabolome Extraction Method Applications? a) Diagnostics Olivier & Loots. (2012). Journal of Microbiological Methods Patent: PCT/IB2012051995 b) Drug Resistance Rif: Olivier & Loots. (2012). Omics. a) Virulence Meissner-Roloff et al. (2012). Metabolomics (hyper vs hypo) Swanepoel et al. (2013). Metabolomics (ESX-1) b) Growth Loots et al. (2013). Metabolomics (ESX-3) Loots et al., submitted, Metabolomics (Iron) c) Drug Resistance INH: Loots 2014. Antimicrobial Agents and Chemotherapy b) Diagnostics Patient sputum samples Patient samples 1. Sputum: • Homogenization: Schoeman et al. 2012. Journal of Microbiological Methods • Applied to 95 Patient sputum samples: du Preez & Loots. (2013). Tuberculosis 2. Urine: • Adaptations of Host and Microbe: de Villiers & Loots., Nature Communications (submitted) • Early Prediction of Treatment Outcome: de Villiers et al. 3. Blood: • Sample collection Adaptations of M. tuberculosis to Host - Sputum Glucose oxidase Adaptations of Host to M.tuberculosis - Sputum Adaptations of Host to M. tuberculosis - Urine KAT Adaptations of Host to Microbe in Competition to Survive - Urine Autophagy: • During nutrient deprived conditions/various disease states: vesicles containing long lived cytoplasmic constituents and organelles are fused with lysosomes, and degraded in order to supply components for cell anabolism, by releasing monomeric units of macromolecules: amino acids, fatty acids, DNA breakdown products etc. • Host uses autophagy in it's defence against M. tuberculosis also. • Decreased insulin may also trigger this process. Adaptations of Host to M. tuberculosis - Urine KAT Side effects associated with TB • Epinephine / increase fatty acids: weight loss, insomnia, glucose intolerance (further confirmed by d-gluconic acid δ-lactone) and the link btw TB and diabetes. • Elevated tyrosine and homovanilic acid – precursors to epinephrine (confirming sputum results) • Quinolinic acid: NMDA agonist associated with neurological abnormalities. • Phenylacetic/Phenyllactic acid: vomiting, nausea, diarrhoea, drowsiness, fatigue, loss of appetite and weight loss. Co-treatment Suggestions Melatonin + Anti-TB co-treatment • Eliminates neurotoxicity associated with quinolinic acid • Increase the efficacy of anti-TB drugs and reduce side effects (Loots et al., 2004) BH4+ Anti-TB co-treatment • Treatment for PKU for preventing phenylacetic acid Metformin + Anti-TB co-treatment • Anti-diabetic drug lowering blood sugar levels and increasing insulin sensitivity • In TB patients - ameliorated lung pathology, reduced chronic inflammation, enhanced the specific immune response and increased the efficacy of conventional TB drugs. Metabolomics Contact: Email: [email protected] Meet the eminent gathering once again at Lung & Respiratory Care -2016 Manchester, UK August 01 - 03, 2016 Lung & Respiratory Care – 2016 Website: lung.conferenceseries.com