Rational Framework for the Design of Trp- and Arg-Rich Peptide Antibiotics Against Multidrug-Resistant Bacteria
The threat of antibiotic resistance warrants the discovery of agents with novel antimicrobial mechanisms. Antimicrobial peptides (AMPs) directly disrupting bacterial membranes may overcome resistance to traditional antibiotics. AMP development for clinical use has been mostly limited to topical appl...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2022.889791/full |
| _version_ | 1818213081488031744 |
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| author | Wenyu Xiang Patrice Clemenza Jessie Klousnitzer Jespar Chen Weiheng Qin Stephanie Tristram-Nagle Yohei Doi Y. Peter Di Berthony Deslouches |
| author_facet | Wenyu Xiang Patrice Clemenza Jessie Klousnitzer Jespar Chen Weiheng Qin Stephanie Tristram-Nagle Yohei Doi Y. Peter Di Berthony Deslouches |
| author_sort | Wenyu Xiang |
| collection | DOAJ |
| description | The threat of antibiotic resistance warrants the discovery of agents with novel antimicrobial mechanisms. Antimicrobial peptides (AMPs) directly disrupting bacterial membranes may overcome resistance to traditional antibiotics. AMP development for clinical use has been mostly limited to topical application to date. We developed a rational framework for systematically addressing this challenge using libraries composed of 86 novel Trp- and Arg-rich engineered peptides tested against clinical strains of the most common multidrug-resistant bacteria known as ESKAPE pathogens. Structure-function correlations revealed minimum lengths (as low as 16 residues) and Trp positioning for maximum antibacterial potency with mean minimum inhibitory concentration (MIC) of 2–4 μM and corresponding negligible toxicity to mammalian cells. Twelve peptides were selected based on broad-spectrum activity against both gram-negative and -positive bacteria and <25% toxicity to mammalian cells at maximum test concentrations. Most of the selected PAX remained active against the colistin-resistant clinical strains. Of the selected peptides, the shortest (the 16-residue E35) was further investigated for antibacterial mechanism and proof-of-concept in vivo efficacy. E35 killed an extensively-resistant isolate of Pseudomonas aeruginosa (PA239 from the CDC, also resistant to colistin) by irreversibly disrupting the cell membranes as shown by propidium iodide incorporation, using flow cytometry and live cell imaging. As proof of concept, in vivo toxicity studies showed that mice tolerated a systemic dose of up to 30 mg/kg peptide and were protected with a single 5 mg/kg intravenous (IV) dose against an otherwise lethal intraperitoneal injection of PA239. Efficacy was also demonstrated in an immune-compromised Klebsiella pneumoniae infection model using a daily dose of 4mg/kg E35 systemically for 2 days. This framework defines the determinants of efficacy of helical AMPs composed of only cationic and hydrophobic amino acids and provides a path for a potential departure from the restriction to topical use of AMPs toward systemic application. |
| first_indexed | 2024-12-12T05:58:37Z |
| format | Article |
| id | doaj.art-609951ce00f9461daa7345404a47a07c |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-12T05:58:37Z |
| publishDate | 2022-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-609951ce00f9461daa7345404a47a07c2022-12-22T00:35:30ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-05-011310.3389/fmicb.2022.889791889791Rational Framework for the Design of Trp- and Arg-Rich Peptide Antibiotics Against Multidrug-Resistant BacteriaWenyu Xiang0Patrice Clemenza1Jessie Klousnitzer2Jespar Chen3Weiheng Qin4Stephanie Tristram-Nagle5Yohei Doi6Y. Peter Di7Berthony Deslouches8Department of Environmental and Occupational Health, School of Public Health, University of Pittsburgh, Pittsburgh, PA, United StatesDepartment of Environmental and Occupational Health, School of Public Health, University of Pittsburgh, Pittsburgh, PA, United StatesDepartment of Environmental and Occupational Health, School of Public Health, University of Pittsburgh, Pittsburgh, PA, United StatesBiological Physics Group, Department of Physics, Carnegie Mellon University, Pittsburgh, PA, United StatesBiological Physics Group, Department of Physics, Carnegie Mellon University, Pittsburgh, PA, United StatesBiological Physics Group, Department of Physics, Carnegie Mellon University, Pittsburgh, PA, United StatesDivision of Infectious Diseases, University of Pittsburgh School of Medicine, Pittsburgh, PA, United StatesDepartment of Environmental and Occupational Health, School of Public Health, University of Pittsburgh, Pittsburgh, PA, United StatesDepartment of Environmental and Occupational Health, School of Public Health, University of Pittsburgh, Pittsburgh, PA, United StatesThe threat of antibiotic resistance warrants the discovery of agents with novel antimicrobial mechanisms. Antimicrobial peptides (AMPs) directly disrupting bacterial membranes may overcome resistance to traditional antibiotics. AMP development for clinical use has been mostly limited to topical application to date. We developed a rational framework for systematically addressing this challenge using libraries composed of 86 novel Trp- and Arg-rich engineered peptides tested against clinical strains of the most common multidrug-resistant bacteria known as ESKAPE pathogens. Structure-function correlations revealed minimum lengths (as low as 16 residues) and Trp positioning for maximum antibacterial potency with mean minimum inhibitory concentration (MIC) of 2–4 μM and corresponding negligible toxicity to mammalian cells. Twelve peptides were selected based on broad-spectrum activity against both gram-negative and -positive bacteria and <25% toxicity to mammalian cells at maximum test concentrations. Most of the selected PAX remained active against the colistin-resistant clinical strains. Of the selected peptides, the shortest (the 16-residue E35) was further investigated for antibacterial mechanism and proof-of-concept in vivo efficacy. E35 killed an extensively-resistant isolate of Pseudomonas aeruginosa (PA239 from the CDC, also resistant to colistin) by irreversibly disrupting the cell membranes as shown by propidium iodide incorporation, using flow cytometry and live cell imaging. As proof of concept, in vivo toxicity studies showed that mice tolerated a systemic dose of up to 30 mg/kg peptide and were protected with a single 5 mg/kg intravenous (IV) dose against an otherwise lethal intraperitoneal injection of PA239. Efficacy was also demonstrated in an immune-compromised Klebsiella pneumoniae infection model using a daily dose of 4mg/kg E35 systemically for 2 days. This framework defines the determinants of efficacy of helical AMPs composed of only cationic and hydrophobic amino acids and provides a path for a potential departure from the restriction to topical use of AMPs toward systemic application.https://www.frontiersin.org/articles/10.3389/fmicb.2022.889791/fullantimicrobial peptidesantibiotic resistancepeptide antibioticsmultidrug resistanceESKAPE pathogensantimicrobial agents |
| spellingShingle | Wenyu Xiang Patrice Clemenza Jessie Klousnitzer Jespar Chen Weiheng Qin Stephanie Tristram-Nagle Yohei Doi Y. Peter Di Berthony Deslouches Rational Framework for the Design of Trp- and Arg-Rich Peptide Antibiotics Against Multidrug-Resistant Bacteria Frontiers in Microbiology antimicrobial peptides antibiotic resistance peptide antibiotics multidrug resistance ESKAPE pathogens antimicrobial agents |
| title | Rational Framework for the Design of Trp- and Arg-Rich Peptide Antibiotics Against Multidrug-Resistant Bacteria |
| title_full | Rational Framework for the Design of Trp- and Arg-Rich Peptide Antibiotics Against Multidrug-Resistant Bacteria |
| title_fullStr | Rational Framework for the Design of Trp- and Arg-Rich Peptide Antibiotics Against Multidrug-Resistant Bacteria |
| title_full_unstemmed | Rational Framework for the Design of Trp- and Arg-Rich Peptide Antibiotics Against Multidrug-Resistant Bacteria |
| title_short | Rational Framework for the Design of Trp- and Arg-Rich Peptide Antibiotics Against Multidrug-Resistant Bacteria |
| title_sort | rational framework for the design of trp and arg rich peptide antibiotics against multidrug resistant bacteria |
| topic | antimicrobial peptides antibiotic resistance peptide antibiotics multidrug resistance ESKAPE pathogens antimicrobial agents |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2022.889791/full |
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