The lysosomotropic drug LeuLeu-OMe induces lysosome disruption and autophagy-independent cell death in Trypanosoma brucei
Background: Trypanosoma brucei is a blood-borne, protozoan parasite that causes African sleeping sickness in humans and nagana in animals. The current chemotherapy relies on only a handful of drugs that display undesirable toxicity, poor efficacy and drug-resistance. In this study, we explored t...
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Shared Science Publishers OG
2015-07-01
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| Series: | Microbial Cell |
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| Online Access: | http://microbialcell.com/researcharticles/the-lysosomotropic-drug-leuleu-ome-induces-lysosome-disruption-and-autophagy-independent-cell-death-in-trypanosoma-brucei/ |
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| author | Hazel Xinyu Koh Htay Mon Aye Kevin S. W. Tan Cynthia Y. He |
| author_facet | Hazel Xinyu Koh Htay Mon Aye Kevin S. W. Tan Cynthia Y. He |
| author_sort | Hazel Xinyu Koh |
| collection | DOAJ |
| description | Background: Trypanosoma brucei is a blood-borne, protozoan parasite that causes African sleeping sickness in humans and nagana in animals. The current chemotherapy relies on only a handful of drugs that display undesirable toxicity, poor efficacy and drug-resistance. In this study, we explored the use of lysosomotropic drugs to induce bloodstream form T. brucei cell death via lysosome destabilization. Methods: We measured drug concentrations that inhibit cell proliferation by 50% (IC50) for several compounds, chosen based on their lysosomotropic effects previously reported in Plasmodium falciparum. The lysosomal effects and cell death induced by L-leucyl-L-leucyl methyl ester (LeuLeu-OMe) were further analyzed by flow cytometry and immunofluorescence analyses of different lysosomal markers. The effect of autophagy in LeuLeu-OMe-induced lysosome destabilization and cytotoxicity was also investigated in control and autophagy-deficient cells. Results: LeuLeu-OMe was selected for detailed analyses due to its strong inhibitory profile against T. brucei with minimal toxicity to human cell lines in vitro. Time-dependent immunofluorescence studies confirmed an effect of LeuLeu-OMe on the lysosome. LeuLeu-OMe-induced cytotoxicity was also found to be dependent on the acidic pH of the lysosome. Although an increase in autophagosomes was observed upon LeuLeu-OMe treatment, autophagy was not required for the cell death induced by LeuLeu-OMe. Necrosis appeared to be the main cause of cell death upon LeuLeu-OMe treatment. Conclusions: LeuLeu-OMe is a lysosomotropic agent capable of destabilizing lysosomes and causing necrotic cell death in bloodstream form of T. brucei. |
| format | Article |
| id | doaj-art-d4f78e7fa55241c392deaf91ddacd07f |
| institution | DOAJ |
| issn | 2311-2638 |
| language | English |
| publishDate | 2015-07-01 |
| publisher | Shared Science Publishers OG |
| record_format | Article |
| series | Microbial Cell |
| spelling | doaj-art-d4f78e7fa55241c392deaf91ddacd07f2025-08-20T02:57:39ZengShared Science Publishers OGMicrobial Cell2311-26382015-07-012828829810.15698/mic2015.08.217123455678The lysosomotropic drug LeuLeu-OMe induces lysosome disruption and autophagy-independent cell death in Trypanosoma bruceiHazel Xinyu Koh0Htay Mon Aye1Kevin S. W. Tan2Cynthia Y. He3Department of Biological Sciences, National University of Singapore.Department of Biological Sciences, National University of Singapore.Department of Microbiology, National University of Singapore.Department of Biological Sciences, National University of Singapore.Background: Trypanosoma brucei is a blood-borne, protozoan parasite that causes African sleeping sickness in humans and nagana in animals. The current chemotherapy relies on only a handful of drugs that display undesirable toxicity, poor efficacy and drug-resistance. In this study, we explored the use of lysosomotropic drugs to induce bloodstream form T. brucei cell death via lysosome destabilization. Methods: We measured drug concentrations that inhibit cell proliferation by 50% (IC50) for several compounds, chosen based on their lysosomotropic effects previously reported in Plasmodium falciparum. The lysosomal effects and cell death induced by L-leucyl-L-leucyl methyl ester (LeuLeu-OMe) were further analyzed by flow cytometry and immunofluorescence analyses of different lysosomal markers. The effect of autophagy in LeuLeu-OMe-induced lysosome destabilization and cytotoxicity was also investigated in control and autophagy-deficient cells. Results: LeuLeu-OMe was selected for detailed analyses due to its strong inhibitory profile against T. brucei with minimal toxicity to human cell lines in vitro. Time-dependent immunofluorescence studies confirmed an effect of LeuLeu-OMe on the lysosome. LeuLeu-OMe-induced cytotoxicity was also found to be dependent on the acidic pH of the lysosome. Although an increase in autophagosomes was observed upon LeuLeu-OMe treatment, autophagy was not required for the cell death induced by LeuLeu-OMe. Necrosis appeared to be the main cause of cell death upon LeuLeu-OMe treatment. Conclusions: LeuLeu-OMe is a lysosomotropic agent capable of destabilizing lysosomes and causing necrotic cell death in bloodstream form of T. brucei.http://microbialcell.com/researcharticles/the-lysosomotropic-drug-leuleu-ome-induces-lysosome-disruption-and-autophagy-independent-cell-death-in-trypanosoma-brucei/Trypanosoma bruceiLeuLeu-OMelysosomelysosomotropicnecrosisautophagy |
| spellingShingle | Hazel Xinyu Koh Htay Mon Aye Kevin S. W. Tan Cynthia Y. He The lysosomotropic drug LeuLeu-OMe induces lysosome disruption and autophagy-independent cell death in Trypanosoma brucei Microbial Cell Trypanosoma brucei LeuLeu-OMe lysosome lysosomotropic necrosis autophagy |
| title | The lysosomotropic drug LeuLeu-OMe induces lysosome disruption and autophagy-independent cell death in Trypanosoma brucei |
| title_full | The lysosomotropic drug LeuLeu-OMe induces lysosome disruption and autophagy-independent cell death in Trypanosoma brucei |
| title_fullStr | The lysosomotropic drug LeuLeu-OMe induces lysosome disruption and autophagy-independent cell death in Trypanosoma brucei |
| title_full_unstemmed | The lysosomotropic drug LeuLeu-OMe induces lysosome disruption and autophagy-independent cell death in Trypanosoma brucei |
| title_short | The lysosomotropic drug LeuLeu-OMe induces lysosome disruption and autophagy-independent cell death in Trypanosoma brucei |
| title_sort | lysosomotropic drug leuleu ome induces lysosome disruption and autophagy independent cell death in trypanosoma brucei |
| topic | Trypanosoma brucei LeuLeu-OMe lysosome lysosomotropic necrosis autophagy |
| url | http://microbialcell.com/researcharticles/the-lysosomotropic-drug-leuleu-ome-induces-lysosome-disruption-and-autophagy-independent-cell-death-in-trypanosoma-brucei/ |
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