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Experimental Therapeutics

Art-175 Is a Highly Efficient Antibacterial against Multidrug-Resistant Strains and Persisters of Pseudomonas aeruginosa

Yves Briers, Maarten Walmagh, Barbara Grymonprez, Manfred Biebl, Jean-Paul Pirnay, Valerie Defraine, Jan Michiels, William Cenens, Abram Aertsen, Stefan Miller, Rob Lavigne
Yves Briers
aLaboratory of Gene Technology, Department Biosystems, KU Leuven, Heverlee, Belgium
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Maarten Walmagh
aLaboratory of Gene Technology, Department Biosystems, KU Leuven, Heverlee, Belgium
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Barbara Grymonprez
aLaboratory of Gene Technology, Department Biosystems, KU Leuven, Heverlee, Belgium
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Manfred Biebl
bLisando GmbH, Regensburg, Germany
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Jean-Paul Pirnay
cLaboratory for Molecular and Cellular Technology, Queen Astrid Military Hospital, Brussels, Belgium
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Valerie Defraine
dCentre of Microbial and Plant Genetics, Department Microbial and Molecular Systems, KU Leuven, Heverlee, Belgium
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Jan Michiels
dCentre of Microbial and Plant Genetics, Department Microbial and Molecular Systems, KU Leuven, Heverlee, Belgium
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William Cenens
eLaboratory of Food Microbiology, Department of Microbial and Molecular Systems, KU Leuven, Heverlee, Belgium
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Abram Aertsen
eLaboratory of Food Microbiology, Department of Microbial and Molecular Systems, KU Leuven, Heverlee, Belgium
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Stefan Miller
bLisando GmbH, Regensburg, Germany
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Rob Lavigne
aLaboratory of Gene Technology, Department Biosystems, KU Leuven, Heverlee, Belgium
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DOI: 10.1128/AAC.02668-14
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  • FIG 1
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    FIG 1

    Bactericidal effects of SMAP-29, KZ144, Art-085, and Art-175 against P. aeruginosa PAO1. A cell suspension was treated with equimolar amounts of SMAP-29, KZ144, Art-085, and Art-175 (3 μM) in the absence and presence of 0.125 or 0.5 mM EDTA. Bacterial reduction after 1 h is expressed in log10 units relative to the untreated control. Mean values (± standard deviation [SD]) are shown. An asterisk indicates when the detection limit (5.15 log units) was reached.

  • FIG 2
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    FIG 2

    Rapid mode of action of Art-175. (A) P. aeruginosa PAO1 is rapidly lysed upon contact with Art-175 (25× MIC). Shortly after adding Art-175, a hole is the cell wall is punctured and the cytoplasmic membrane increasingly bulges until the cell undergoes abrupt lysis (see also Movie S1 in the supplemental material). (B) A 30-min time course shows that P. aeruginosa PAO1 treated with 25× MIC Art-175 is reduced by approximately 3 log units in the first 5 min, increasing to 4 log units after 30 min. Mean values (± SD) are shown.

  • FIG 3
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    FIG 3

    Resistance development against Art-175 and ciprofloxacin. A laboratory strain of clinical origin (P. aeruginosa PAO1) (A), an environmental strain (P. aeruginosa Br257) (B), and a multidrug-resistant strain (P. aeruginosa Br776) (C) were serially exposed to subinhibitory concentrations to select for decreased susceptibility. Over 20 passages, the MIC of Art-175 (◊) increased 2-fold, whereas the MIC of ciprofloxacin (☐) increased 16- (PAO1), 64- (Br257), or 4-fold (Br776).

  • FIG 4
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    FIG 4

    Persister killing by Art-175. The bactericidal effects of Art-175 at 10× and 30× MIC on isolated persister fractions of P. aeruginosa PA14 (light gray) and PA1255 (dark gray) are compared to those of ciprofloxacin at 10× MIC and 30× MIC, in both the absence and the presence of 0.5 mM EDTA. Controls include 0.5 mM EDTA and ofloxacin (10× MIC and 30× MIC). Mean values (± standard error of the mean [SEM]) are shown for at least three independent replicates.

Tables

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  • TABLE 1

    MICs of Art-085 and Art-175a

    EDTA concn (mM) or parameterMIC (μg/ml) for:
    Art-085Art-175
    01810
    0.1252010
    0.25166
    0.5104
    164
    242
    442
    MIC50b84
    MIC901510
    • ↵a Art-175 (10 μg/ml) has an improved inhibitory activity against P. aeruginosa PAO1 compared to that of Art-085 (18 μg/ml). From 0.25 mM EDTA, MICs further decrease to 4 and 2 μg/ml for Art-085 and Art-175, respectively.

    • ↵b The MICs at which growth of 50% (MIC50) or 90% (MIC90) of the isolates are inhibited, are shown.

Additional Files

  • Figures
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  • Supplemental material

    Files in this Data Supplement:

    • Supplemental file 1 -

      Supplemental Figures S1 to S3 and Tables S1 to S3.

      PDF, 658K

    • Supplemental file 2 -

      Supplemental Movie S1: time-lapse series during exposure of P. aeruginosa to Art-175 (25× MIC, 0.5 mM EDTA). Recording started ~1 min after addition of the protein (scale bar = 4 µm; 5 s per frame; total duration, 21 min 5 s).

      AVI, 8.0M

    • Supplemental file 3 -

      Supplemental Movie S2: time-lapse series during exposure of P. aeruginosa to Art-175 (30× MIC, 0.5 mM EDTA). Recording started ~1 min after addition of the protein (scale bar = 4 µm; 5 s per frame; total duration, 5 min 25 s).

      AVI, 2.5M

    • Supplemental file 4 -

      Supplemental Movie S3: time-lapse series during exposure of P. aeruginosa to SMAP-29 (25× MIC, 0.5 mM EDTA). Recording started ~1 min after addition of the protein (scale bar = 4 µm; 2 min per frame; total duration, 3 h 48 min).

      AVI, 3.8M

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Art-175 Is a Highly Efficient Antibacterial against Multidrug-Resistant Strains and Persisters of Pseudomonas aeruginosa
Yves Briers, Maarten Walmagh, Barbara Grymonprez, Manfred Biebl, Jean-Paul Pirnay, Valerie Defraine, Jan Michiels, William Cenens, Abram Aertsen, Stefan Miller, Rob Lavigne
Antimicrobial Agents and Chemotherapy Jun 2014, 58 (7) 3774-3784; DOI: 10.1128/AAC.02668-14

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Art-175 Is a Highly Efficient Antibacterial against Multidrug-Resistant Strains and Persisters of Pseudomonas aeruginosa
Yves Briers, Maarten Walmagh, Barbara Grymonprez, Manfred Biebl, Jean-Paul Pirnay, Valerie Defraine, Jan Michiels, William Cenens, Abram Aertsen, Stefan Miller, Rob Lavigne
Antimicrobial Agents and Chemotherapy Jun 2014, 58 (7) 3774-3784; DOI: 10.1128/AAC.02668-14
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