3616479052629d305f46250c7895f44f pharmaceutics_14_02169_v3.pdf f4c294d5f60817309e947e71b0a75e33d8e42ce7 pharmaceutics_14_02169_v3.pdf 184ab590e4d5e0eed5871e69e1d31b038300c8abb80c4bac7900d2b4c71f8e83 pharmaceutics_14_02169_v3.pdf Title: Antimicrobial Peptides Can Generate Tolerance by Lag and Interfere with Antimicrobial Therapy Subject: Antimicrobial peptides (AMPs) are widely distributed molecules secreted mostly by cells of the innate immune system to prevent bacterial proliferation at the site of infection. As with classic antibiotics, continued treatment with AMPs can create resistance in bacteria. However, whether AMPs can generate tolerance as an intermediate stage towards resistance is not known. Here, we show that the treatment of Escherichia coli with different AMPs induces tolerance by lag, particularly for those peptides that have internal targets. This tolerance can be detected as different morphological and physiological changes, which depend on the type of peptide molecule the bacterium has been exposed to. In addition, we show that AMP tolerance can also affect antibiotic treatment. The genomic sequencing of AMP-tolerant strains shows that different mutations alter membrane composition, DNA replication, and translation. Some of these mutations have also been observed in antibiotic-resistant strains, suggesting that AMP tolerance could be a relevant step in the development of antibiotic resistance. Monitoring AMP tolerance is relevant vis-á-vis the eventual therapeutic use of AMPs and because cross-tolerance might favor the emergence of resistance against conventional antibiotic treatments. Keywords: tolerance; antimicrobial peptides; antimicrobial peptide LL-37; pleurocidin; polymyxin B; dermaseptin Author: Daniel Sandín, Javier Valle, Jordi Morata, David Andreu and Marc Torrent Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Oct 25 08:51:43 2022 CEST ModDate: Tue Oct 25 09:59:08 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 10 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 5851038 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- CIQLLF+VnURWPalladioL Type 1 Custom yes yes no 10 0 KGSZMU+VnURWPalladioL-Italic Type 1 Custom yes yes no 15 0 MGVEUC+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 YJJSBV+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 GFXHXR+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 WLENKM+VnURWPalladioL-Bold Type 1 Custom yes yes no 32 0 IAFAOG+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 51 0 XIRVNA+EURB10 Type 1 Builtin yes yes no 79 0 TDUDTH+CMSY10 Type 1 Builtin yes yes no 83 0 ZXDKIP+EURM10 Type 1 Builtin yes yes no 87 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 102 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 106 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 110 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 124 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 128 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 132 0 BCDJEE+PalatinoLinotype-BoldItalic TrueType WinAnsi yes yes no 136 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 160 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 164 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 168 0 BCDJEE+PalatinoLinotype-BoldItalic TrueType WinAnsi yes yes no 172 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-09-02 02:28:44 CEST RepresentationInformation: pharmaceutics_14_02169_v3.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-09-01 09:12:24 CEST Size: 5851038 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 314 FreeObjects: 3 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Materials Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Peptide Tolerant Strain Development and Data Processing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Minimum Inhibitory Concentration (MIC) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Killing Curve Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: DNA Purification and Genome Sequencing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Children: Item: Title: Antimicrobial Peptides Generate Tolerance by Lag Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Tolerance in AMPs Can Affect Antimicrobial Treatments Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: The Tolerance Mutational Landscape Is Diverse Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Info: Title: Antimicrobial Peptides Can Generate Tolerance by Lag and Interfere with Antimicrobial Therapy Author: Daniel Sandín, Javier Valle, Jordi Morata, David Andreu and Marc Torrent Subject: Antimicrobial peptides (AMPs) are widely distributed molecules secreted mostly by cells of the innate immune system to prevent bacterial proliferation at the site of infection. As with classic antibiotics, continued treatment with AMPs can create resistance in bacteria. However, whether AMPs can generate tolerance as an intermediate stage towards resistance is not known. Here, we show that the treatment of Escherichia coli with different AMPs induces tolerance by lag, particularly for those peptides that have internal targets. This tolerance can be detected as different morphological and physiological changes, which depend on the type of peptide molecule the bacterium has been exposed to. In addition, we show that AMP tolerance can also affect antibiotic treatment. The genomic sequencing of AMP-tolerant strains shows that different mutations alter membrane composition, DNA replication, and translation. Some of these mutations have also been observed in antibiotic-resistant strains, suggesting that AMP tolerance could be a relevant step in the development of antibiotic resistance. Monitoring AMP tolerance is relevant vis-á-vis the eventual therapeutic use of AMPs and because cross-tolerance might favor the emergence of resistance against conventional antibiotic treatments. 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