High-Throughput Chemical Screening for Antivirulence Developmental Phenotypes in Trypanosoma brucei
| العنوان: | High-Throughput Chemical Screening for Antivirulence Developmental Phenotypes in Trypanosoma brucei |
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| المؤلفون: | Macgregor, Paula, Ivens, Alasdair, Shave, Steven, Collie, Iain, Gray, David, Auer, Manfred, Matthews, Keith R |
| المصدر: | Eukaryotic Cell Macgregor, P, Ivens, A, Shave, S, Collie, I, Gray, D, Auer, M & Matthews, K R 2014, ' High throughput chemical screening for anti-virulence developmental phenotypes in Trypanosoma brucei ', Eukaryotic Cell, vol. 13, no. 3, pp. 412-426 . https://doi.org/10.1128/EC.00335-13Test |
| بيانات النشر: | American Society for Microbiology, 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | Small Molecule Libraries, Phenotype, Virulence, Trypanosoma brucei brucei, Protozoan Proteins, Articles, RNA, Messenger, High-Throughput Screening Assays |
| الوصف: | In the bloodstream of mammalian hosts, the sleeping sickness parasite, Trypanosoma brucei, exists as a proliferative slender form or a nonproliferative, transmissible, stumpy form. The transition between these developmental forms is controlled by a density-dependent mechanism that is important for the parasite's infection dynamics, immune evasion via ordered antigenic variation, and disease transmissibility. However, stumpy formation has been lost in most laboratory-adapted trypanosome lines, generating monomorphic parasites that proliferate uncontrolled as slender forms in vitro and in vivo. Nonetheless, these forms are readily amenable to cell culture and high-throughput screening for trypanocidal lead compounds. Here, we have developed and exploited a high-throughput screen for developmental phenotypes using a transgenic monomorphic cell line expressing a reporter under the regulation of gene control signals from the stumpy-specific molecule PAD1. Using a whole-cell fluorescence-based assay to screen over 6,000 small molecules from a kinase-focused compound library, small molecules able to activate stumpy-specific gene expression and proliferation arrest were assayed in a rapid assay format. Independent follow-up validation identified one hit able to induce modest, yet specific, changes in mRNA expression indicative of a partial differentiation to stumpy forms in monomorphs. Further, in pleomorphs this compound induced a stumpy-like phenotype, entailing growth arrest, morphological changes, PAD1 expression, and enhanced differentiation to procyclic forms. This not only provides a potential tool compound for the further understanding of stumpy formation but also demonstrates the use of high-throughput screening in the identification of compounds able to induce specific phenotypes, such as differentiation, in African trypanosomes. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1535-9786 1535-9778 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=pmid_dedup__::84e95ae47e0894bb8e78527f14808b05Test http://europepmc.org/articles/PMC3957582Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.pmid.dedup....84e95ae47e0894bb8e78527f14808b05 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15359786 15359778 |
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