High-throughput screen for novel antimicrobials using a whole animal infection model.

The nematode Caenorhabditis elegans is a unique whole animal model system for identifying small molecules with in vivo anti-infective properties. C. elegans can be infected with a broad range of human pathogens, including Enterococcus faecalis, an important human nosocomial pathogen. Here, we describe an automated, high-throughput screen of 37,200 compounds and natural product extracts for those that enhance survival of C. elegans infected with E. faecalis. Using a robot to dispense live, infected animals into 384-well plates and automated microscopy and image analysis, we identified 28 compounds and extracts not previously reported to have antimicrobial properties, including six structural classes that cure infected C. elegans animals but do not affect the growth of the pathogen in vitro, thus acting by a mechanism of action distinct from antibiotics currently in clinical use.

[1]  D. Bumann.  Has nature already identified all useful antibacterial targets? , 2008, Current opinion in microbiology.

[2]  Martin Serrano,et al.  Nucleic Acids Research Advance Access published October 18, 2007 ChemBank: a small-molecule screening and , 2007 .

[3]  F. Ausubel,et al.  Models of Caenorhabditis elegans infection by bacterial and fungal pathogens. , 2008, Methods in molecular biology.

[4]  G. Marshall,et al.  The biology and future prospects of antivirulence therapies , 2008, Nature Reviews Microbiology.

[5]  Audrey White,et al.  Automated, quantitative screening assay for antiangiogenic compounds using transgenic zebrafish. , 2007, Cancer research.

[6]  L. Buck,et al.  An antidepressant that extends lifespan in adult Caenorhabditis elegans , 2007, Nature.

[7]  Jon Clardy,et al.  Small-molecule Inhibitors of Pseudomonas aeruginosa Biofilm Development , 2012, The Meducator.

[8]  Anne E Carpenter,et al.  CellProfiler: image analysis software for identifying and quantifying cell phenotypes , 2006, Genome Biology.

[9]  Frederick M Ausubel,et al.  Identification of novel antimicrobials using a live-animal infection model , 2006, Proceedings of the National Academy of Sciences.

[10]  Trevor C. Y. Kwok,et al.  A small-molecule screen in C. elegans yields a new calcium channel antagonist , 2006, Nature.

[11]  Theresa Stiernagle.  Maintenance of C. elegans. , 2006, WormBook : the online review of C. elegans biology.

[12]  A. Marra.  Targeting Virulence for Antibacterial Chemotherapy , 2006 .

[13]  A. Marra.  Targeting virulence for antibacterial chemotherapy: identifying and characterising virulence factors for lead discovery. , 2006, Drugs in R&D.

[14]  S. Walker,et al.  Discovery of O-GlcNAc transferase inhibitors. , 2005, Journal of the American Chemical Society.

[15]  L. Silver.  A retrospective on the failures and successes of antibacterial drug discovery. , 2005, IDrugs : the investigational drugs journal.

[16]  M. Schmid.  Seeing is believing: the impact of structural genomics on antimicrobial drug discovery , 2004, Nature Reviews Microbiology.

[17]  A. Boggs,et al.  Antibacterial drug discovery: is small pharma the solution? , 2004, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[18]  S. Projan,et al.  Antibacterial drug discovery: is it all downhill from here? , 2004, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[19]  C. Walsh.  Opinion — anti-infectives: Where will new antibiotics come from? , 2003, Nature Reviews Microbiology.

[20]  Anders Olsen,et al.  An automated high-throughput assay for survival of the nematode Caenorhabditis elegans. , 2003, Free Radical Biology & Medicine.

[21]  Cori Bargmann,et al.  SEK‐1 MAPKK mediates Ca2+ signaling to determine neuronal asymmetric development in Caenorhabditis elegans , 2002, EMBO reports.

[22]  Thomas D. Y. Chung,et al.  A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays , 1999, Journal of biomolecular screening.

[23]  F. Ausubel,et al.  Killing of Caenorhabditis elegans by Pseudomonas aeruginosa used to model mammalian bacterial pathogenesis. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Strome,et al.  Characterization of a germ-line proliferation mutation in C. elegans. , 1992, Development.