title: Inhibition of Pseudomonas aeruginosa by Peptide-Conjugated Phosphorodiamidate Morpholino Oligomers.
abstract: Pseudomonas aeruginosais a highly virulent, multidrug-resistant pathogen that causes significant morbidity and mortality in hospitalized patients and is particularly devastating in patients with cystic fibrosis. Increasing antibiotic resistance coupled with decreasing numbers of antibiotics in the developmental pipeline demands novel antibacterial approaches. Here, we tested peptide-conjugated phosphorodiamidate morpholino oligomers (PPMOs), which inhibit translation of complementary mRNA from specific, essential genes inP. aeruginosaPPMOs targeted toacpP,lpxC, andrpsJ, inhibitedP. aeruginosagrowth in many clinical strains and activity of PPMOs could be enhanced 2- to 8-fold by the addition of polymyxin B nonapeptide at subinhibitory concentrations. The PPMO targetingacpPwas also effective at preventingP. aeruginosaPAO1 biofilm formation and at reducing existing biofilms. Importantly, treatment with various combinations of a PPMO and a traditional antibiotic demonstrated synergistic growth inhibition, the most effective of which was the PPMO targetingrpsJwith tobramycin. Furthermore, treatment ofP. aeruginosaPA103-infected mice with PPMOs targetingacpP,lpxC, orrpsJsignificantly reduced the bacterial burden in the lungs at 24 h by almost 3 logs. Altogether, this study demonstrates that PPMOs targeting the essential genesacpP,lpxC, orrpsJinP. aeruginosaare highly effective at inhibiting growthin vitroandin vivoThese data suggest that PPMOs alone or in combination with antibiotics represent a novel approach to addressing the problems associated with rapidly increasing antibiotic resistance inP. aeruginosa.
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: James J
authors.initials: JJ
authors.lastname: Howard
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Sturge
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Dina A
authors.initials: DA
authors.lastname: Moustafa
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Seth M
authors.initials: SM
authors.lastname: Daly
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Kimberly R
authors.initials: KR
authors.lastname: Marshall-Batty
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Christina F
authors.initials: CF
authors.lastname: Felder
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Danniel
authors.initials: D
authors.lastname: Zamora
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Marium
authors.initials: M
authors.lastname: Yabe-Gill
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Labandeira-Rey
authors.affiliation: Sarepta Therapeutics, Cambridge, Massachusetts, USA.
authors.forename: Stacey M
authors.initials: SM
authors.lastname: Bailey
authors.affiliation: Sarepta Therapeutics, Cambridge, Massachusetts, USA.|Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Wong
authors.affiliation: Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Joanna B
authors.initials: JB
authors.lastname: Goldberg
authors.affiliation: Department of Microbiology, Oregon State University, Corvallis, Oregon, USA.
authors.forename: Bruce L
authors.initials: BL
authors.lastname: Geller
authors.affiliation: Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA david.greenberg@utsouthwestern.edu.|Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Greenberg
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D018441:Biofilms / Q000187:drug effects / Q000254:growth & development; D054889:Fatty Acid Synthase, Type II / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008826:Microbial Sensitivity Tests; D058990:Molecular Targeted Therapy; D060172:Morpholinos / Q000737:chemistry / Q000494:pharmacology*; D016376:Oligonucleotides, Antisense / Q000737:chemistry / Q000494:pharmacology*; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D011552:Pseudomonas Infections / Q000188:drug therapy* / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D012269:Ribosomal Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D060172:Morpholinos; D016376:Oligonucleotides, Antisense; D010455:Peptides; D012269:Ribosomal Proteins; D000581:Amidohydrolases; C514484:LpxC deacetylase, Pseudomonas; D054889:Fatty Acid Synthase, Type II
keywords: PPMO; Pseudomonas aeruginosa; antibiotic resistance; antimicrobial agents; antisense; experimental therapeutics; phosphorodiamidate morpholino oligomer