709. High-throughput Screening (HTS) for anti-infectives using a Whole Animal Infection Model
Session: Poster Abstract Session: Antimicrobials: Novel Agents
Friday, October 4, 2013
Room: The Moscone Center: Poster Hall C
Posters
  • IDSA poster 2013.pdf (820.7 kB)
  • Background: Treatment of bacterial infections has become increasingly complicated due to emergence of resistance mechanisms against established antibiotics. This in turn necessitates identification of novel anti-infectives with alternative modes of action. We have developed a C. elegans based HTS model to screen chemical libraries for anti-infectives against methicillin-resistant Staphylococcus aureus(MRSA) infection. This whole animal model not only facilitates identification of molecules with anti-microbial, anti-virulence or immunomodulatory properties but also tests for their toxicity in the same system.

    Methods: C. elegans were infected with MRSA and simultaneously treated with test compounds. The efficacy of the compounds was determined after a four-day incubation period. The state of the worm was evaluated using Sytox, an agent that stains the nucleic acids of cells with compromised membranes. Worms that are alive and cured of the infection do not stain, whereas dead worms fluoresce. Automated microscopy and high content imaging analysis enabled identification of chemical hits.

    Results: We have developed an automated, HTS assay that uses large particle sorter and liquid handling robotics to dispense compounds, bacteria and live C. elegansinto 384-wells plates. A comparison of vancomycin (positive control) and DMSO (negative control) provided a Z’ factor of 0.80, indicating a good quality assay with wide separation between signal and background. A screen of 19,344 compounds yielded 143 hits which included compounds whose anti-infective properties against MRSA was previously unknown.

    Conclusion: HT screening of the whole animal infection model can identify compounds that cure MRSA infection. Follow-up studies will determine whether these compounds target bacterial virulence or stimulate host immunity

    Raj Mohan Raja Muthiah, Ph.D.1, Beth Fuchs, Ph.D.1, Jonah Larkins-Ford2, Annie Connery2, Frederick Ausubel, PhD3 and Eleftherios Mylonakis, MD, PhD1, (1)Infectious Diseases, Rhode Island Hospital, Brown University, Providence, RI, (2)Massachusetts General Hospital, Boston, MA, (3)Massachusetts General Hosp, Harvard Med School, Boston, MA

    Disclosures:

    R. M. Raja Muthiah, None

    B. Fuchs, None

    J. Larkins-Ford, None

    A. Connery, None

    F. Ausubel, None

    E. Mylonakis, None

    Findings in the abstracts are embargoed until 12:01 a.m. PST, Oct. 2nd with the exception of research findings presented at the IDWeek press conferences.