An in situ high-throughput screen identifies inhibitors of intracellular Burkholderia pseudomallei with therapeutic efficacy.

Autor:	Bulterys PL; University of California, Los Angeles-California Institute of Technology Medical Scientist Training Program, David Geffen School of Medicine, University of California, Los Angeles, CA 90095.; Molecular Biology Institute, University of California, Los Angeles, CA 90095.; Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095., Toesca IJ; Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095., Norris MH; Department of Infectious Diseases and Immunology, College of Veterinary Medicine, University of Florida, Gainesville, FL 32611.; Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611., Maloy JP; Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095., Fitz-Gibbon ST; Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, CA 90095., France B; California NanoSystems Institute, University of California, Los Angeles, CA 90095., Toffig B; California NanoSystems Institute, University of California, Los Angeles, CA 90095., Morselli M; Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, CA 90095., Somprasong N; Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611.; Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32611., Pellegrini M; Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, CA 90095.; California NanoSystems Institute, University of California, Los Angeles, CA 90095.; Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095., Schweizer HP; Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611.; Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32611., Tuanyok A; Department of Infectious Diseases and Immunology, College of Veterinary Medicine, University of Florida, Gainesville, FL 32611.; Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611., Damoiseaux R; California NanoSystems Institute, University of California, Los Angeles, CA 90095.; Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095., French CT; Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095; ctfrench@ucla.edu jfmiller@g.ucla.edu.; California NanoSystems Institute, University of California, Los Angeles, CA 90095., Miller JF; Molecular Biology Institute, University of California, Los Angeles, CA 90095; ctfrench@ucla.edu jfmiller@g.ucla.edu.; Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095.; California NanoSystems Institute, University of California, Los Angeles, CA 90095.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2019 Sep 10; Vol. 116 (37), pp. 18597-18606. Date of Electronic Publication: 2019 Aug 22.
DOI:	10.1073/pnas.1906388116
Abstrakt:	Burkholderia pseudomallei ( Bp ) and Burkholderia mallei ( Bm ) are Tier-1 Select Agents that cause melioidosis and glanders, respectively. These are highly lethal human infections with limited therapeutic options. Intercellular spread is a hallmark of Burkholderia pathogenesis, and its prominent ties to virulence make it an attractive therapeutic target. We developed a high-throughput cell-based phenotypic assay and screened ∼220,000 small molecules for their ability to disrupt intercellular spread by Burkholderia thailandensis , a closely related BSL-2 surrogate. We identified 268 hits, and cross-species validation found 32 hits that also disrupt intercellular spread by Bp and/or Bm Among these were a fluoroquinolone analog, which we named burkfloxacin (BFX), which potently inhibits growth of intracellular Burkholderia , and flucytosine (5-FC), an FDA-approved antifungal drug. We found that 5-FC blocks the intracellular life cycle at the point of type VI secretion system 5 (T6SS-5)-mediated cell-cell spread. Bacterial conversion of 5-FC to 5-fluorouracil and subsequently to fluorouridine monophosphate is required for potent and selective activity against intracellular Burkholderia In a murine model of fulminant respiratory melioidosis, treatment with BFX or 5-FC was significantly more effective than ceftazidime, the current antibiotic of choice, for improving survival and decreasing bacterial counts in major organs. Our results demonstrate the utility of cell-based phenotypic screening for Select Agent drug discovery and warrant the advancement of BFX and 5-FC as candidate therapeutics for melioidosis in humans. Competing Interests: The authors declare no conflict of interest. (Copyright © 2019 the Author(s). Published by PNAS.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu