Inhibiting the stringent response blocks Mycobacterium tuberculosis entry into quiescence and reduces persistence

Noton K Dutta; Lee G Klinkenberg; Maria-Jesus Vazquez; Delfina Segura-Carro; Gonzalo Colmenarejo; Fernando Ramon; Beatriz Rodriguez-Miquel; Lydia Mata-Cantero; Esther Porras-De Francisco; Yu-Min Chuang; Harvey Rubin; Jae Jin Lee; Hyungjin Eoh; Joel S Bader; Esther Perez-Herran; Alfonso Mendoza-Losana; Petros C Karakousis

doi:10.1126/sciadv.aav2104

Inhibiting the stringent response blocks Mycobacterium tuberculosis entry into quiescence and reduces persistence

Sci Adv. 2019 Mar 20;5(3):eaav2104. doi: 10.1126/sciadv.aav2104. eCollection 2019 Mar.

Authors

Noton K Dutta¹, Lee G Klinkenberg¹, Maria-Jesus Vazquez², Delfina Segura-Carro³, Gonzalo Colmenarejo^{4

5}, Fernando Ramon⁴, Beatriz Rodriguez-Miquel³, Lydia Mata-Cantero³, Esther Porras-De Francisco³, Yu-Min Chuang¹, Harvey Rubin⁶, Jae Jin Lee⁷, Hyungjin Eoh⁷, Joel S Bader⁸, Esther Perez-Herran³, Alfonso Mendoza-Losana³, Petros C Karakousis^{1

9}

Affiliations

¹ Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
² Medical Department, GlaxoSmithKline, Tres Cantos, Madrid, Spain.
³ Diseases of the Developing World, GlaxoSmithKline, Tres Cantos, Madrid, Spain.
⁴ Molecular Discovery Research, GlaxoSmithKline, Tres Cantos, Madrid, Spain.
⁵ Biostatistics and Bioinformatics Unit, IMDEA Food Institute, Madrid, Spain.
⁶ Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
⁷ Department of Molecular Microbiology and Immunology, Keck School of Medicine of USC, Los Angeles, CA, USA.
⁸ Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁹ Department of International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA.

Abstract

The stringent response enables Mycobacterium tuberculosis (Mtb) to shut down its replication and metabolism under various stresses. Here we show that Mtb lacking the stringent response enzyme Rel_Mtb was unable to slow its replication rate during nutrient starvation. Metabolomics analysis revealed that the nutrient-starved rel_Mtb -deficient strain had increased metabolism similar to that of exponentially growing wild-type bacteria in nutrient-rich broth, consistent with an inability to enter quiescence. Deficiency of rel_Mtb increased the susceptibility of mutant bacteria to killing by isoniazid during nutrient starvation and in the lungs of chronically infected mice. We screened a pharmaceutical library of over 2 million compounds for inhibitors of Rel_Mtb and showed that the lead compound X9 was able to directly kill nutrient-starved M. tuberculosis and enhanced the killing activity of isoniazid. Inhibition of Rel_Mtb is a promising approach to target M. tuberculosis persisters, with the potential to shorten the duration of TB treatment.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Bacterial Proteins / antagonists & inhibitors
Bacterial Proteins / chemistry
Bacterial Proteins / genetics*
Crystallography, X-Ray
DNA Replication / drug effects
Escherichia coli Proteins / antagonists & inhibitors
Escherichia coli Proteins / chemistry
GTP Pyrophosphokinase / antagonists & inhibitors
GTP Pyrophosphokinase / chemistry
Humans
Intracellular Signaling Peptides and Proteins / antagonists & inhibitors*
Intracellular Signaling Peptides and Proteins / chemistry
Intracellular Signaling Peptides and Proteins / genetics
Isoniazid / chemistry
Isoniazid / pharmacology
Mice
Mycobacterium tuberculosis / chemistry
Mycobacterium tuberculosis / drug effects*
Mycobacterium tuberculosis / genetics
Mycobacterium tuberculosis / pathogenicity
Protein Conformation
Small Molecule Libraries / chemistry
Small Molecule Libraries / pharmacology
Tuberculosis / drug therapy
Tuberculosis / genetics*
Tuberculosis / microbiology
Tuberculosis / pathology

Substances

Bacterial Proteins
Escherichia coli Proteins
Intracellular Signaling Peptides and Proteins
RshA protein, Mycobacterium tuberculosis
Small Molecule Libraries
GTP Pyrophosphokinase
relA protein, E coli
Isoniazid

Abstract

Publication types

MeSH terms

Substances

Grants and funding