The Tyrosine Kinase Inhibitor Gefitinib Restricts Mycobacterium tuberculosis Growth through Increased Lysosomal Biogenesis and Modulation of Cytokine Signaling

ACS Infect Dis. 2017 Aug 11;3(8):564-574. doi: 10.1021/acsinfecdis.7b00046. Epub 2017 Jun 5.

Abstract

Host-directed therapeutics have the potential to combat the global tuberculosis pandemic. We previously identified gefitinib, an inhibitor of EGFR, as a potential host-targeted therapeutic effective against Mycobacterium tuberculosis infection of macrophages and mice. Here we examine the functional consequences of gefitinib treatment on M. tuberculosis infected macrophages. Using phosphoproteomic and transcriptional profiling, we identify two mechanisms by which gefitinib influences macrophage responses to infection to affect cytokine responses and limit replication of M. tuberculosis in macrophages. First, we find that gefitinib treatment of M. tuberculosis infected macrophages inhibits STAT3, a transcription factor known to repress effective immune responses to M. tuberculosis in vivo. Second, we find that gefitinib treatment of M. tuberculosis infected macrophages leads to increased expression of genes involved in lysosomal biogenesis and function and an increase of functional lysosomes in gefitinib treated cells. Furthermore, we show that gefitinib treatment increases the targeting of bacteria to lysosomes, providing an explanation for the cell intrinsic effects of gefitinib treatment on M. tuberculosis infection. Our data provide novel insights into the effects of gefitinib on mammalian cells and into the possible roles for EGFR signaling in macrophages.

Keywords: Mycobacterium tuberculosis; epidermal growth factor receptor (EGFR); host-directed therapeutic; lysosomal biogenesis; signal transducer and activator of transcription 3 (STAT3).

MeSH terms

  • Animals
  • Antitubercular Agents / pharmacology*
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / metabolism
  • Bone Marrow Cells / microbiology
  • Cytokines / antagonists & inhibitors
  • Cytokines / genetics
  • Cytokines / metabolism
  • ErbB Receptors / antagonists & inhibitors
  • ErbB Receptors / genetics
  • ErbB Receptors / metabolism
  • Gefitinib
  • Gene Expression Regulation
  • Lysosomes / drug effects*
  • Lysosomes / microbiology
  • Macrophages / drug effects*
  • Macrophages / metabolism
  • Macrophages / microbiology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mycobacterium tuberculosis / drug effects*
  • Mycobacterium tuberculosis / growth & development
  • Mycobacterium tuberculosis / pathogenicity
  • Organelle Biogenesis
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism
  • Primary Cell Culture
  • Protein Kinase Inhibitors / pharmacology*
  • Proteomics / methods
  • Quinazolines / pharmacology*
  • STAT3 Transcription Factor / antagonists & inhibitors*
  • STAT3 Transcription Factor / genetics
  • STAT3 Transcription Factor / metabolism
  • Signal Transduction

Substances

  • Antitubercular Agents
  • Cytokines
  • Phosphoproteins
  • Protein Kinase Inhibitors
  • Quinazolines
  • STAT3 Transcription Factor
  • Stat3 protein, mouse
  • EGFR protein, mouse
  • ErbB Receptors
  • Gefitinib