Insights into antitrypanosomal drug mode-of-action from cytology-based profiling
| Autor: | Thomas, James A., Baker, Nicola, Hutchinson, Sebastian, Dominicus, Caia, Trenaman, Anna, Glover, Lucy, Alsford, Sam, Horn, David |
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| Přispěvatelé: | London School of Hygiene and Tropical Medicine (LSHTM), University of York [York, UK], Institut Pasteur [Paris], The Francis Crick Institute [London], University of Dundee, We thank Philippa Radley for assistance with assembly of tagging constructs, Richard Wheeler (Uni. of Oxford) for advice on ImageJ analysis and Achim Schnaufer (Uni. of Edinburgh) for the ATP-synthase antibody, Institut Pasteur [Paris] (IP) |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Physiology
[SDV]Life Sciences [q-bio] RC955-962 Melarsoprol MESH: Melarsoprol Biochemistry Arctic medicine. Tropical medicine Medicine and Health Sciences Cell Cycle and Cell Division MESH: Trypanocidal Agents Energy-Producing Organelles Protozoans Chromosome Biology Pharmaceutics Eukaryota MESH: Mitochondrial Proteins Trypanocidal Agents Mitochondria Electrophysiology Nucleic acids Cell Processes MESH: Pentamidine Cellular Structures and Organelles Public aspects of medicine RA1-1270 Research Article Trypanosoma MESH: Mitochondria Trypanosoma brucei brucei Mitosis Suramin Bioenergetics DNA replication Membrane Potential MESH: Cell Biology Mitochondrial Proteins Drug Therapy parasitic diseases Trypanosoma Brucei Genetics Humans MESH: Suramin Pentamidine MESH: Humans MESH: Nifurtimox Organisms Biology and Life Sciences MESH: Trypanosoma brucei brucei Cell Biology DNA MESH: Mitosis Parasitic Protozoans Kinetoplasts Trypanosomiasis African MESH: Trypanosomiasis African Nifurtimox Lysosomes Trypanosoma Brucei Gambiense MESH: Lysosomes |
| Zdroj: | PLoS Neglected Tropical Diseases PLoS Neglected Tropical Diseases, Public Library of Science, 2018, 12 (11), pp.e0006980. ⟨10.1371/journal.pntd.0006980⟩ PLoS Neglected Tropical Diseases, Vol 12, Iss 11, p e0006980 (2018) PLoS Neglected Tropical Diseases, 2018, 12 (11), pp.e0006980. ⟨10.1371/journal.pntd.0006980⟩ |
| ISSN: | 1935-2727 1935-2735 |
| DOI: | 10.1371/journal.pntd.0006980⟩ |
| Popis: | Chemotherapy continues to have a major impact on reducing the burden of disease caused by trypanosomatids. Unfortunately though, the mode-of-action (MoA) of antitrypanosomal drugs typically remains unclear or only partially characterised. This is the case for four of five current drugs used to treat Human African Trypanosomiasis (HAT); eflornithine is a specific inhibitor of ornithine decarboxylase. Here, we used a panel of T. brucei cellular assays to probe the MoA of the current HAT drugs. The assays included DNA-staining followed by microscopy and quantitative image analysis, or flow cytometry; terminal dUTP nick end labelling to monitor mitochondrial (kinetoplast) DNA replication; antibody-based detection of sites of nuclear DNA damage; and fluorescent dye-staining of mitochondria or lysosomes. We found that melarsoprol inhibited mitosis; nifurtimox reduced mitochondrial protein abundance; pentamidine triggered progressive loss of kinetoplast DNA and disruption of mitochondrial membrane potential; and suramin inhibited cytokinesis. Thus, current antitrypanosomal drugs perturb distinct and specific cellular compartments, structures or cell cycle phases. Further exploiting the findings, we show that putative mitogen-activated protein-kinases contribute to the melarsoprol-induced mitotic defect, reminiscent of the mitotic arrest associated signalling cascade triggered by arsenicals in mammalian cells, used to treat leukaemia. Thus, cytology-based profiling can rapidly yield novel insight into antitrypanosomal drug MoA. Author summary African trypanosomes cause devastating and lethal diseases in humans and livestock. These parasites are transmitted among mammals by tsetse flies and circulate and grow in blood and tissue fluids. There are several drugs available to treat patients but, despite their use for many decades, we know relatively little about how they work. We reasoned that exposure of trypanosomes to each drug, followed by microscopic examination of cellular structures, would reveal the major cellular compartments, structures or growth phases affected. For example, we examined two major DNA structures, and cellular compartments known as mitochondria. We found that two drugs thought to act in mitochondria did indeed disrupt this compartment, but in completely different ways. Another drug stopped cell growth at a specific point in the cycle. An arsenic-based drug, related to anti-leukaemia drugs, perturbed the nuclear DNA division cycle, indicating that arsenicals may kill parasites and cancer cells by similar mechanisms. Thus, the ‘chemical-biology’ profiles we observe illuminate distinct killing mechanisms. A similar approach can now be used to assess new drugs, and the insights may help to develop improved anti-parasite therapies. |
| Databáze: | OpenAIRE |
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