b27492658c8bb9ad81057aa63a70e890 biomedicines_a2022v10n2p386.pdf bbfc11ebe6fd397a96502dae5a40d6c821c345be biomedicines_a2022v10n2p386.pdf 51509e78d9c226ea828cca5fba860a5d2cf3c2d5e8e8d1da8108f6eb31450b37 biomedicines_a2022v10n2p386.pdf Title: In Vivo Evaluation of ECP Peptide Analogues for the Treatment of Acinetobacter baumannii Infection Subject: Antimicrobial peptides (AMPs) are alternative therapeutics to traditional antibiotics against bacterial resistance. Our previous work identified an antimicrobial region at the N-terminus of the eosinophil cationic protein (ECP). Following structure-based analysis, a 30mer peptide (ECPep-L) was designed that combines antimicrobial action against Gram-negative species with lipopolysaccharides (LPS) binding and endotoxin-neutralization activities. Next, analogues that contain non-natural amino acids were designed to increase serum stability. Here, two analogues were selected for in vivo assays: the all-D version (ECPep-D) and the Arg to Orn version that incorporates a D-amino acid at position 2 (ECPep-2D-Orn). The peptide analogues retained high LPS-binding and anti-endotoxin activities. The peptides efficacy was tested in a murine acute infection model of Acinetobacter baumannii. Results highlighted a survival rate above 70% following a 3-day supervision with a single administration of ECPep-D. Moreover, in both ECPep-D and ECPep-2D-Orn peptide-treated groups, clinical symptoms improved significantly and the tissue infection was reduced to equivalent levels to mice treated with colistin, used as a last resort in the clinics. Moreover, treatment drastically reduced serum levels of TNF- inflammation marker within the first 8 h. The present results support ECP-derived peptides as alternative candidates for the treatment of acute infections caused by Gram-negative bacteria. Keywords: ECP; AMPs; infection; murine model; Gram-negative bacteria; LPS Author: Jiarui Li, Guillem Prats-Ejarque, Marc Torrent, David Andreu, Klaus Brandenburg, Pablo Fernández-Millán and Ester Boix Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Fri Feb 11 10:07:36 2022 CET ModDate: Thu Feb 24 10:32:09 2022 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 20 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 14810575 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HCRKXL+VnURWPalladioL Type 1 Custom yes yes no 10 0 VVSYYB+EURM10 Type 1 Builtin yes yes no 15 0 COQOXH+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 GGUFYQ+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 VHWTCC+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 JQTLGG+URWPalladioL-BoldItal Type 1 Custom yes yes no 32 0 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Identity-H yes no yes 211 0 CIDFont+F1 CID TrueType Identity-H yes no yes 219 0 CIDFont+F2 CID TrueType Identity-H yes no yes 227 0 CIDFont+F3 CID TrueType Identity-H yes no yes 278 0 CIDFont+F1 CID TrueType Identity-H yes no yes 286 0 CIDFont+F2 CID TrueType Identity-H yes no yes 294 0 CIDFont+F3 CID TrueType Identity-H yes no yes 415 0 CIDFont+F1 CID TrueType Identity-H yes no yes 423 0 CIDFont+F2 CID TrueType Identity-H yes no yes 431 0 CIDFont+F3 CID TrueType Identity-H yes no yes 555 0 CIDFont+F1 CID TrueType Identity-H yes no yes 563 0 CIDFont+F2 CID TrueType Identity-H yes no yes 571 0 CIDFont+F3 CID TrueType Identity-H yes no yes 654 0 CIDFont+F1 CID TrueType Identity-H yes no yes 662 0 CIDFont+F2 CID TrueType Identity-H yes no yes 670 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-03-16 02:27:37 CET RepresentationInformation: biomedicines_a2022v10n2p386.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-02-28 11:52:55 CET Size: 14810575 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 964 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Children: Item: Title: Bacteria Strain, Cells, Mice and Other Materials Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Peptides Synthesis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Circular Dichroism Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Minimum Inhibitory Concentration (MIC) Determination Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: LPS Affinity Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Cytotoxicity Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Tolerance Study of Peptides in Mouse Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Mouse Systemic Infection Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Efficacy Assay of Peptides in Infection Mice Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Evaluation of Body Weight and Clinical Symptoms Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Assay of CFU in Mice Tissues Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Quantification of TNF- and LPS in Mice Serum by ELISA Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Stimulation of Human MNC by LPS and Endotoxin Neutralization Assay Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Children: Item: Title: Design and Structural Characterization of ECP Peptide Analogues Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: In Vitro Activities of ECP Peptides Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: Tolerance of Peptides in Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Item: Title: Murine Acute Infection Model by A. baumannii Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@87f383f Item: Title: Treatment with Peptides Improves the Survival Rate and Relieves Clinical Suffering of Infected Mice Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4eb7f003 Item: Title: ECP Peptides Reduce Bacterial Counts in Mice Organs and TNF- Levels after 8 h of Treatment Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@eafc191 Item: Title: ECP Peptides Reduce TNF- Levels in LPS-Stimulated Cells Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612fc6eb Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1060b431 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@612679d6 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@11758f2a Info: Title: In Vivo Evaluation of ECP Peptide Analogues for the Treatment of Acinetobacter baumannii Infection Author: Jiarui Li, Guillem Prats-Ejarque, Marc Torrent, David Andreu, Klaus Brandenburg, Pablo Fernández-Millán and Ester Boix Subject: Antimicrobial peptides (AMPs) are alternative therapeutics to traditional antibiotics against bacterial resistance. Our previous work identified an antimicrobial region at the N-terminus of the eosinophil cationic protein (ECP). Following structure-based analysis, a 30mer peptide (ECPep-L) was designed that combines antimicrobial action against Gram-negative species with lipopolysaccharides (LPS) binding and endotoxin-neutralization activities. Next, analogues that contain non-natural amino acids were designed to increase serum stability. Here, two analogues were selected for in vivo assays: the all-D version (ECPep-D) and the Arg to Orn version that incorporates a D-amino acid at position 2 (ECPep-2D-Orn). The peptide analogues retained high LPS-binding and anti-endotoxin activities. The peptides efficacy was tested in a murine acute infection model of Acinetobacter baumannii. Results highlighted a survival rate above 70% following a 3-day supervision with a single administration of ECPep-D. Moreover, in both ECPep-D and ECPep-2D-Orn peptide-treated groups, clinical symptoms improved significantly and the tissue infection was reduced to equivalent levels to mice treated with colistin, used as a last resort in the clinics. Moreover, treatment drastically reduced serum levels of TNF- inflammation marker within the first 8 h. The present results support ECP-derived peptides as alternative candidates for the treatment of acute infections caused by Gram-negative bacteria. 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Subtype: Link Rect: 517, 503, 554, 514 Annotation: Subtype: Link Rect: 256, 480, 293, 491 Annotation: Subtype: Link Rect: 459, 457, 496, 468 Annotation: Subtype: Link Rect: 437, 434, 475, 445 Annotation: Subtype: Link Rect: 175, 399, 212, 410 Annotation: Subtype: Link Rect: 218, 399, 254, 410 Annotation: Subtype: Link Rect: 354, 365, 392, 376 Annotation: Subtype: Link Rect: 398, 365, 434, 376 Annotation: Subtype: Link Rect: 95, 331, 133, 341 Annotation: Subtype: Link Rect: 139, 331, 175, 341 Annotation: Subtype: Link Rect: 251, 308, 289, 318 Annotation: Subtype: Link Rect: 295, 308, 331, 318 Checksum: 9b37ae4f Type: CRC32 Checksum: b27492658c8bb9ad81057aa63a70e890 Type: MD5 Checksum: bbfc11ebe6fd397a96502dae5a40d6c821c345be Type: SHA-1