Insights into antitrypanosomal drug mode-of-action from cytology-based profiling.
James A ThomasNicola BakerSebastian HutchinsonCaia DominicusAnna TrenamanLucy GloverSam AlsfordDavid HornPublished in: PLoS neglected tropical diseases (2018)
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.
Keyphrases
- cell cycle
- flow cytometry
- oxidative stress
- dna damage
- drug induced
- cell proliferation
- single molecule
- high resolution
- high throughput
- endothelial cells
- high grade
- quantum dots
- living cells
- fine needle aspiration
- cell free
- single cell
- diabetic rats
- cell death
- radiation therapy
- risk assessment
- high speed
- nucleic acid
- small molecule
- dna repair
- climate change
- adverse drug
- microbial community
- highly efficient
- stress induced