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A 2-pyridone-amide inhibitor targets the glucose metabolism pathway of Chlamydia trachomatis.

Engström P, Krishnan KS, Ngyuen BD, Chorell E, Normark J, Silver J, Bastidas RJ, Welch MD, Hultgren SJ, Wolf-Watz H, Valdivia RH, Almqvist F, Bergström S - MBio (2014)

Bottom Line: Consistent with an effect on G-6P metabolism, treatment with KSK120 blocked glycogen accumulation.Interestingly, KSK120 did not affect Escherichia coli or the host cell.Thus, KSK120 may be a useful tool to study chlamydial glucose metabolism and has the potential to be used in the treatment of C. trachomatis infections.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Umeå University, Umeå, Sweden Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden Umeå Centre for Microbial Research (UCMR), Umeå University, Umeå, Sweden Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.

No MeSH data available.


Related in: MedlinePlus

Compound KSK120 has selective anti-C. trachomatis activity that blocks its infectivity. (A) Compounds synthesized with substituent variations decorating the 2-pyridone central scaffolds. See Table S1 in the supplemental material for a complete list of compounds tested. (B) HeLa cells infected with C. trachomatis serovar LGV-2 were treated for the entire infection with 10 µM KSK120 or DMSO, fixed with methanol at 44 hour postinfection (hpi), and stained with primary antibodies toward major outer membrane protein (MOMP) (red) and heat shock protein 60 (Hsp60) (green). DAPI was used to detect bacterial and host DNA (blue). Confocal laser scanning microscopy was used to obtain the images. (C) HeLa cells infected with C. trachomatis LGV-2 were treated with KSK120 or DMSO, and at 44 hpi, infectious progeny were collected for reinfection of new HeLa cells. DMSO treatment results were set to a value of 100. Data presented were acquired from an experiment performed in triplicate. (D) Generation of infectious EB (elementary body) progeny of indicated C. trachomatis serovars in the presence of 10 µM KSK120 (44 hpi) or the corresponding amount of DMSO. Presented data are the means of the results of three experiments. (E) Representative transmission electron micrographs of cells infected with C. trachomatis LGV-2 treated with 10 µM KSK120 or the corresponding amount of DMSO (44 hpi). (F) Growth and biofilm formation in the presence of KSK120. Results were quantified relative to nontreated E. coli results. Representative experiments were performed in triplicate. In all figures, error bars indicate standard deviations (SD).
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fig1: Compound KSK120 has selective anti-C. trachomatis activity that blocks its infectivity. (A) Compounds synthesized with substituent variations decorating the 2-pyridone central scaffolds. See Table S1 in the supplemental material for a complete list of compounds tested. (B) HeLa cells infected with C. trachomatis serovar LGV-2 were treated for the entire infection with 10 µM KSK120 or DMSO, fixed with methanol at 44 hour postinfection (hpi), and stained with primary antibodies toward major outer membrane protein (MOMP) (red) and heat shock protein 60 (Hsp60) (green). DAPI was used to detect bacterial and host DNA (blue). Confocal laser scanning microscopy was used to obtain the images. (C) HeLa cells infected with C. trachomatis LGV-2 were treated with KSK120 or DMSO, and at 44 hpi, infectious progeny were collected for reinfection of new HeLa cells. DMSO treatment results were set to a value of 100. Data presented were acquired from an experiment performed in triplicate. (D) Generation of infectious EB (elementary body) progeny of indicated C. trachomatis serovars in the presence of 10 µM KSK120 (44 hpi) or the corresponding amount of DMSO. Presented data are the means of the results of three experiments. (E) Representative transmission electron micrographs of cells infected with C. trachomatis LGV-2 treated with 10 µM KSK120 or the corresponding amount of DMSO (44 hpi). (F) Growth and biofilm formation in the presence of KSK120. Results were quantified relative to nontreated E. coli results. Representative experiments were performed in triplicate. In all figures, error bars indicate standard deviations (SD).

Mentions: On the basis of previous observations that 2-pyridone carboxylic acids can block biofilm formation of E. coli (13, 14), as well as our recent findings showing that 2-pyridones have the ability to block infection of the human intracellular pathogen Listeria monocytogenes (unpublished observation), we were prompted to investigate a potential effect on chlamydial infectivity. Therefore, we screened a library of 61 2-pyridone compounds with various substitutions around a 2-pyridone central scaffold (Fig. 1A; see also Table S1 in the supplemental material) for their ability to affect intracellular infection by C. trachomatis serovar LGV-L2. To identify compounds that inhibit the production of infectious progeny, we assessed each compound for its effect on bacterial distribution within the inclusion. We chose this assay because we previously found that salicylidene acylhydrazide compounds, which cause a reduction in the yields of infectious progeny, alter the normally homogeneous distribution of bacteria within the inclusion to a nonuniform and patchy distribution (15). Each compound, or a dimethyl sulfoxide (DMSO) control, was added to HeLa cells immediately after infection, and the distribution of bacteria within inclusions was assessed at 44 h postinfection (hpi) using confocal fluorescence microscopy. As expected, bacteria were homogenously dispersed inside the inclusions of DMSO-treated control cells (Fig. 1B, left). In contrast, 3 compounds caused bacteria to be heterogeneously dispersed, whereas 8 compounds caused moderate heterogeneous dispersion and 12 were toxic to HeLa cells (see Table S1). The remaining compounds had no effects on either C. trachomatis distribution or the host cell.


A 2-pyridone-amide inhibitor targets the glucose metabolism pathway of Chlamydia trachomatis.

Engström P, Krishnan KS, Ngyuen BD, Chorell E, Normark J, Silver J, Bastidas RJ, Welch MD, Hultgren SJ, Wolf-Watz H, Valdivia RH, Almqvist F, Bergström S - MBio (2014)

Compound KSK120 has selective anti-C. trachomatis activity that blocks its infectivity. (A) Compounds synthesized with substituent variations decorating the 2-pyridone central scaffolds. See Table S1 in the supplemental material for a complete list of compounds tested. (B) HeLa cells infected with C. trachomatis serovar LGV-2 were treated for the entire infection with 10 µM KSK120 or DMSO, fixed with methanol at 44 hour postinfection (hpi), and stained with primary antibodies toward major outer membrane protein (MOMP) (red) and heat shock protein 60 (Hsp60) (green). DAPI was used to detect bacterial and host DNA (blue). Confocal laser scanning microscopy was used to obtain the images. (C) HeLa cells infected with C. trachomatis LGV-2 were treated with KSK120 or DMSO, and at 44 hpi, infectious progeny were collected for reinfection of new HeLa cells. DMSO treatment results were set to a value of 100. Data presented were acquired from an experiment performed in triplicate. (D) Generation of infectious EB (elementary body) progeny of indicated C. trachomatis serovars in the presence of 10 µM KSK120 (44 hpi) or the corresponding amount of DMSO. Presented data are the means of the results of three experiments. (E) Representative transmission electron micrographs of cells infected with C. trachomatis LGV-2 treated with 10 µM KSK120 or the corresponding amount of DMSO (44 hpi). (F) Growth and biofilm formation in the presence of KSK120. Results were quantified relative to nontreated E. coli results. Representative experiments were performed in triplicate. In all figures, error bars indicate standard deviations (SD).
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Related In: Results  -  Collection

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fig1: Compound KSK120 has selective anti-C. trachomatis activity that blocks its infectivity. (A) Compounds synthesized with substituent variations decorating the 2-pyridone central scaffolds. See Table S1 in the supplemental material for a complete list of compounds tested. (B) HeLa cells infected with C. trachomatis serovar LGV-2 were treated for the entire infection with 10 µM KSK120 or DMSO, fixed with methanol at 44 hour postinfection (hpi), and stained with primary antibodies toward major outer membrane protein (MOMP) (red) and heat shock protein 60 (Hsp60) (green). DAPI was used to detect bacterial and host DNA (blue). Confocal laser scanning microscopy was used to obtain the images. (C) HeLa cells infected with C. trachomatis LGV-2 were treated with KSK120 or DMSO, and at 44 hpi, infectious progeny were collected for reinfection of new HeLa cells. DMSO treatment results were set to a value of 100. Data presented were acquired from an experiment performed in triplicate. (D) Generation of infectious EB (elementary body) progeny of indicated C. trachomatis serovars in the presence of 10 µM KSK120 (44 hpi) or the corresponding amount of DMSO. Presented data are the means of the results of three experiments. (E) Representative transmission electron micrographs of cells infected with C. trachomatis LGV-2 treated with 10 µM KSK120 or the corresponding amount of DMSO (44 hpi). (F) Growth and biofilm formation in the presence of KSK120. Results were quantified relative to nontreated E. coli results. Representative experiments were performed in triplicate. In all figures, error bars indicate standard deviations (SD).
Mentions: On the basis of previous observations that 2-pyridone carboxylic acids can block biofilm formation of E. coli (13, 14), as well as our recent findings showing that 2-pyridones have the ability to block infection of the human intracellular pathogen Listeria monocytogenes (unpublished observation), we were prompted to investigate a potential effect on chlamydial infectivity. Therefore, we screened a library of 61 2-pyridone compounds with various substitutions around a 2-pyridone central scaffold (Fig. 1A; see also Table S1 in the supplemental material) for their ability to affect intracellular infection by C. trachomatis serovar LGV-L2. To identify compounds that inhibit the production of infectious progeny, we assessed each compound for its effect on bacterial distribution within the inclusion. We chose this assay because we previously found that salicylidene acylhydrazide compounds, which cause a reduction in the yields of infectious progeny, alter the normally homogeneous distribution of bacteria within the inclusion to a nonuniform and patchy distribution (15). Each compound, or a dimethyl sulfoxide (DMSO) control, was added to HeLa cells immediately after infection, and the distribution of bacteria within inclusions was assessed at 44 h postinfection (hpi) using confocal fluorescence microscopy. As expected, bacteria were homogenously dispersed inside the inclusions of DMSO-treated control cells (Fig. 1B, left). In contrast, 3 compounds caused bacteria to be heterogeneously dispersed, whereas 8 compounds caused moderate heterogeneous dispersion and 12 were toxic to HeLa cells (see Table S1). The remaining compounds had no effects on either C. trachomatis distribution or the host cell.

Bottom Line: Consistent with an effect on G-6P metabolism, treatment with KSK120 blocked glycogen accumulation.Interestingly, KSK120 did not affect Escherichia coli or the host cell.Thus, KSK120 may be a useful tool to study chlamydial glucose metabolism and has the potential to be used in the treatment of C. trachomatis infections.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, Umeå University, Umeå, Sweden Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå University, Umeå, Sweden Umeå Centre for Microbial Research (UCMR), Umeå University, Umeå, Sweden Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.

No MeSH data available.


Related in: MedlinePlus