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Principal Investigator: DAVID Branch MOODY
Organization: BRIGHAM AND WOMEN'S HOSPITAL
Fiscal Year: 2020
Award: $223,437
Funding agency: National Institute of Allergy and Infectious Diseases
The Metabolomics Core (B) is comprised of a Biosafety Level 3 suite for handling infectious samples,
massively parallel detection of mycobacterial metabolites using Time of Flight mass spectrometry (Agilent
Accurate Mass ToF 6230), specialized resources for identifying known mycobacterial compounds, and
analytical capabilities to discover previously unknown compounds (Agilent Accurate Mass 6520, QTof,
Thermo LXQ Advantage 2 Dimensional Ion Trap with MSn). The integrated technologies were assembled by
the Moody laboratory and supported a Biomarker Discover Initiative of the Broad Institute, NIH U19 and ROl
Projects. Dataflow involves receipt of mycobacteria (Project 3) or patient samples (Project 4), from which
total metabolites are extracted and sterilized in organic solvents. Tissue or mycobacterial extracts enter into
a liquid chromatography-mass spectrometry system, which was specifically designed to broadly detect the
highly diverse and hydrophobic compounds in mycobacteria. In the first phase of "whole organism" analysis,
the platform rapidly detects triplicate intensity values for ~10,000 distinct compounds in each sample. By
aligning large datasets derived from different patients, clinical isolates or genetically engineered bacteria, in-house-designed software pipeline identifies all compounds that are changed at statistically significant levels.
In a second, targeted phase, all changed compounds are ranked by biological or quantitative criteria to
define compounds of interest, whose structures are solved by comparing their masses to the literature
(MycoMass) and in-house (MycoMap) databases or are solved through collisional mass spectrometry. This
system has discovered previously unknown compounds, identified strain-specific mycobacterial biomarkers
in vitro and from tissues and identified lipids changed after gene deletion. This overview describes
expansion of the substantial existing core facilities, including a new generation of high accuracy mass
spectrometry and expansion of mycobacterial databases, as well as use of the Core to discover biomarkers
in drug-resistant or latent mycobacteria or biomarkers of infection.
Terms: <2-dimensional><Bacteria><Bio-Informatics><Bioinformatics><Biological><Biological Markers><Body Tissues><Cavia><Clinic><Clinical><Core Facility><Data Bases><Databases><Designing computer software><Detection><Disease><Disorder><Drug resistance><Drugs><Gene Deletion><Generations><Genetic Engineering><Genetic Engineering Biotechnology><Genetic Engineering Molecular Biology><Genus Mycobacterium><Guinea Pigs><Guinea Pigs Mammals><Health><Human><Hydrophobicity><In Vitro><Infection><Institutes><Ions><LC/MS><Laboratories><Lipids><Literature><M tb><M tuberculosis><M tuberculosis infection><M. tb><M. tb infection><M. tuberculosis><M. tuberculosis infection><M.tb infection><M.tuberculosis infection><MTB infection><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Medication><Metabolic><Methods><Modern Man><Mycobacterium><Mycobacterium tuberculosis><Mycobacterium tuberculosis (MTB) infection><Mycobacterium tuberculosis infection><NIH><National Institutes of Health><Organic Solvents><Organic solvent product><Patients><Pharmaceutic Preparations><Pharmaceutical Preparations><Phase><Recombinant DNA Technology><Research Resources><Resources><Sampling><Software Design><Structure><System><TB infection><Technology><Tissues><Tuberculosis><United States National Institutes of Health><Whole Organism Analysis><bacterial pathogen><bio-markers><biologic marker><biomarker><biosignature><comparative><data base><data format><design><designing><disseminated TB><disseminated tuberculosis><drug resistant><drug/agent><gene deletion mutation><genetically engineered><human subject><improved><infection due to Mycobacterium tuberculosis><interest><large data sets><large datasets><liquid chromatography mass spectrometry><mass spectrometer><metabolism measurement><metabolome><metabolomics><metabonome><metabonomics><mtb><mycobacterial><pathogenic bacteria><rapid diagnosis><resistance to Drug><resistant to Drug><time of flight mass spectrometry><time use><tuberculosis infection><tuberculous spondyloarthropathy><two-dimensional>