Spatial quantification of drugs in pulmonary tuberculosis lesions by laser capture microdissection liquid chromatography mass spectrometry (LCM-LC/MS)

Matthew Zimmerman, Landry Blanc, Pei Yu Chen, Véronique Dartois, Brendan Prideaux

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Tuberculosis is still a leading cause of morbidity and mortality worldwide. Improvements to existing drug regimens and the development of novel therapeutics are urgently required. The ability of dosed TB drugs to reach and sterilize bacteria within poorly-vascularized necrotic regions (caseum) of pulmonary granulomas is crucial for successful therapeutic intervention. Effective therapeutic regimens must therefore contain drugs with favorable caseum penetration properties. Current LC/MS methods for quantifying drug levels in biological tissues have limited spatial resolution capabilities, making it difficult to accurately determine absolute drug concentrations within small tissue compartments such as those found within necrotic granulomas. Here we present a protocol combining laser capture microdissection (LCM) of pathologically-distinct tissue regions with LC/MS quantification. This technique provides absolute quantification of drugs within granuloma caseum, surrounding cellular lesion and uninvolved lung tissue and, therefore, accurately determines whether bactericidal concentrations are being achieved. In addition to tuberculosis research, the technique has many potential applications for spatially-resolved quantification of drugs in diseased tissues.

Original languageEnglish (US)
Article numbere57402
JournalJournal of Visualized Experiments
Volume2018
Issue number134
DOIs
StatePublished - Apr 18 2018
Externally publishedYes

Keywords

  • Drug distribution
  • Granuloma
  • Issue 134
  • Laser capture microdissection
  • Liquid chromatography mass spectrometry
  • Medicine
  • Necrosis
  • Quantification
  • Tuberculosis

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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