75f9fb17720397f35541408c8ac92df1 pmc_11431785.pdf 3578fdc2ff4f52c7448104b4120d78e64684cc1a pmc_11431785.pdf 1043456a01a8606c80c01b77deefa6cef5d864d0e1b8af897e663ea935cddb5c pmc_11431785.pdf Title: From In Vitro Promise to In Vivo Reality: An Instructive Account of Infection Model Evaluation of Antimicrobial Peptides Subject: Antimicrobial peptides (AMPs) are regarded as a promising alternative to traditional antibiotics in the face of ever-increasing resistance. However, many AMPs fail to progress into clinics due to unexpected difficulties found in preclinical in vivo phases. Our research has focused on crotalicidin (Ctn), an AMP from snake venom, and a fragment thereof, Ctn[15-34], with improved in vitro antimicrobial and anticancer activities and remarkable serum stability. As the retroenantio versions of both AMPs maintained favorable profiles, in this work, we evaluate the in vivo efficacy of both the native-sequence AMPs and their retroenantio counterparts in a murine infection model with Acinetobacter baumannii. A significant reduction in bacterial levels is found in the mice treated with Ctn[15-34]. However, contrary to expectations, the retroenantio analogs either exhibit toxicity or lack efficacy when administered to mice. Our findings underscore the critical importance of in vivo infection model evaluation to fully calibrate the therapeutic potential of AMPs. Keywords: AMPs; topoisomers; retroenantio; infection; murine model; crotalicidin Author: Adam Carrera-Aubesart, Jiarui Li, Estefanía Contreras, Roberto Bello-Madruga, Marc Torrent and David Andreu Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Tue Sep 10 10:55:59 2024 CEST ModDate: Tue Sep 10 11:01:08 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 13 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3282195 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- NXQZWG+EURM10 Type 1 Builtin yes yes yes 10 0 WLENKM+VnURWPalladioL-Bold Type 1 Custom yes yes yes 15 0 ARIQJE+VnURWPalladioL Type 1 Custom yes yes yes 21 0 YYGJDH+URWPalladioL-Bold Type 1 Custom yes yes yes 26 0 ABQLYT+URWPalladioL-Roma Type 1 Custom yes yes yes 32 0 WRGQRU+URWPalladioL-Ital 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0 NOIMPK+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 233 0 NOINLG+PalatinoLinotype TrueType MacRoman yes yes no 236 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-02-05 02:41:52 CET RepresentationInformation: pmc_11431785.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-12-05 02:30:56 CET Size: 3282195 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 465 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results and Discussion Destination: section.2 Children: Item: Title: Peptides Destination: subsection.2.1 Item: Title: Treatment with Ctn[15-34] Reduces Bacterial Levels in Females, Whereas Ctn re Does Not Improve the Survival Rate Destination: subsection.2.2 Item: Title: Treatment with Ctn[15-34] Thrice a Day Lowers Bacterial Load but Does Not Improve Survival Rate Destination: subsection.2.3 Item: Title: Ctn[15-34] Reduces Levels of TNF- in Mice after 8 h and Ctn[15-34] re Increase TNF- Levels in LPS-Stimulated Human MNC Cells Destination: subsection.2.4 Item: Title: Organ Staining Consistent with Bacterial Levels at Each Treatment Destination: subsection.2.5 Item: Title: Materials and Methods Destination: section.3 Children: Item: Title: Mice, Bacterial Strains and Other Material Destination: subsection.3.1 Item: Title: Peptides Destination: subsection.3.2 Item: Title: Minimum Inhibitory Concentration (MIC) Destination: subsection.3.3 Item: Title: Peptide Cytotoxicity Destination: subsection.3.4 Item: Title: Serum Stability Destination: subsection.3.5 Item: Title: Tolerance in Mice Destination: subsection.3.6 Item: Title: Mouse Systemic Infection Destination: subsection.3.7 Item: Title: Efficacy in Mouse Infection Model Destination: subsection.3.8 Item: Title: Evaluation of Body Weight and Clinical Symptoms Destination: subsection.3.9 Item: Title: Evaluation of Colony-Forming Units (CFU) in Mouse Tissues Destination: subsection.3.10 Item: Title: Quantification of TNF- in Mouse Sera by ELISA Destination: subsection.3.11 Item: Title: Stimulation of Human Mononuclear Cells (MNC) by LPS Destination: subsection.3.12 Item: Title: Histopathology, Tissue Gram Stain and Hematoxylin and Eosin Stain Destination: subsection.3.13 Item: Title: Statistical Analysis Destination: subsection.3.14 Item: Title: Conclusions Destination: section.4 Item: Title: References Destination: appendix.A. Info: Title: From In Vitro Promise to In Vivo Reality: An Instructive Account of Infection Model Evaluation of Antimicrobial Peptides Author: Adam Carrera-Aubesart, Jiarui Li, Estefanía Contreras, Roberto Bello-Madruga, Marc Torrent and David Andreu Subject: Antimicrobial peptides (AMPs) are regarded as a promising alternative to traditional antibiotics in the face of ever-increasing resistance. However, many AMPs fail to progress into clinics due to unexpected difficulties found in preclinical in vivo phases. Our research has focused on crotalicidin (Ctn), an AMP from snake venom, and a fragment thereof, Ctn[15-34], with improved in vitro antimicrobial and anticancer activities and remarkable serum stability. As the retroenantio versions of both AMPs maintained favorable profiles, in this work, we evaluate the in vivo efficacy of both the native-sequence AMPs and their retroenantio counterparts in a murine infection model with Acinetobacter baumannii. A significant reduction in bacterial levels is found in the mice treated with Ctn[15-34]. However, contrary to expectations, the retroenantio analogs either exhibit toxicity or lack efficacy when administered to mice. Our findings underscore the critical importance of in vivo infection model evaluation to fully calibrate the therapeutic potential of AMPs. 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