TriTOX: A novel Trichomonas vaginalis assay platform for high-throughput screening of compound libraries
| Autor: | Andrew M. Piggott, Samantha J. Emery-Corbin, Daniel Vuong, Aaron R. Jex, Ernest Lacey, Alexander Y.F. Lam |
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| Rok vydání: | 2021 |
| Předmět: |
Special issue articles on 'Anaerobic Protozoan Pathogens: Drugs
Resistance and New Developments' 0301 basic medicine Sexually transmitted disease Microbial metabolites HTS High-throughput screen Trichomonas Tritrichomonas foetus Infectious and parasitic diseases RC109-216 medicine.disease_cause 0302 clinical medicine MTS Medium-throughput screen Drug-discovery Pharmacology (medical) MIC Minimum inhibitory concentration MetAP2 Methionine aminopeptidase 2 Tritrichomonas Natural products Trichomoniasis biology Mtz Metronidazole Infectious Diseases HIV Human immunodeficiency virus BLAST Basic local alignment search tool STD Sexually-transmitted disease I-TASSER Iterative threading assembly refinement algorithm medicine.drug NI Nitroimidazole 030231 tropical medicine PBS Phosphate buffered saline Microbiology 03 medical and health sciences Entamoeba histolytica Metronidazole parasitic diseases Trichomonas vaginalis medicine Animals Fumagillin MOA Mode of action ADT AutoDockTools Pharmacology DMSO Dimethyl sulfoxide biology.organism_classification medicine.disease High-Throughput Screening Assays 030104 developmental biology PDB Protein data bank Parasitology Giardia lamblia TSA Trichostatin A |
| Zdroj: | International Journal for Parasitology: Drugs and Drug Resistance, Vol 15, Iss, Pp 68-80 (2021) International Journal for Parasitology: Drugs and Drug Resistance |
| ISSN: | 2211-3207 |
| Popis: | Trichomonas vaginalis is a neglected urogenital parasitic protist that causes 170 million cases of trichomoniasis annually, making it the most prevalent non-viral, sexually transmitted disease. Trichomoniasis treatment relies on nitroheterocyclics, such as metronidazole. However, with increasing drug-resistance, there is an urgent need for novel anti-trichomonals. Little progress has been made to translate anti-trichomonal research into commercialised therapeutics, and the absence of a standardised compound-screening platform is the immediate stumbling block for drug-discovery. Herein, we describe a simple, cost-effective growth assay for T. vaginalis and the related Tritrichomonas foetus. Tracking changes in pH were a valid indicator of trichomonad growth (T. vaginalis and T. foetus), allowing development of a miniaturised, chromogenic growth assay based on the phenol red indicator in 96- and 384-well microtiter plate formats. The outputs of this assay can be quantitatively and qualitatively assessed, with consistent dynamic ranges based on Z′ values of 0.741 and 0.870 across medium- and high-throughput formats, respectively. We applied this high-throughput format within the largest pure-compound microbial metabolite screen (812 compounds) for T. vaginalis and identified 43 hit compounds. We compared these identified compounds to mammalian cell lines, and highlighted extensive overlaps between anti-trichomonal and anti-tumour activity. Lastly, observing nanomolar inhibition of T. vaginalis by fumagillin, and noting this compound has reported activity in other protists, we performed in silico analyses of the interaction of fumagillin with its molecular target methionine aminopeptidase 2 for T. vaginalis, Giardia lamblia and Entamoeba histolytica, highlighting potential for fumagillin as a broad-spectrum anti-protistal against microaerophilic protists. Together, this new platform will accelerate drug-discovery efforts, underpin drug-resistance screening in trichomonads, and contributing to a growing body of evidence highlighting the potential of microbial natural products as novel anti-protistals. Graphical abstract Image 1 |
| Databáze: | OpenAIRE |
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