Insights into antitrypanosomal drug mode-of-action from cytology-based profiling

James Thomas, Nicola Baker, Caia Dominicus, Anna Trenaman, Lucy Glover, Sam Alsford, David Horn (Lead / Corresponding author)

Research output: Contribution to journalArticle

78 Downloads (Pure)

Abstract

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.
Original languageEnglish
Article numbere0006980
Pages (from-to)1-19
Number of pages19
JournalPLoS Neglected Tropical Diseases
Volume12
Issue number11
DOIs
Publication statusPublished - 26 Nov 2018

Fingerprint

Cell Biology
Melarsoprol
Kinetoplast DNA
African Trypanosomiasis
Pharmaceutical Preparations
Nifurtimox
Image Cytometry
Staining and Labeling
Eflornithine
Arsenicals
Pentamidine
Suramin
Cytokinesis
Mitochondrial Membrane Potential
Mitochondrial Proteins
Cellular Structures
Lysosomes
Mitogen-Activated Protein Kinases
DNA Replication
Mitochondrial DNA

Keywords

  • Cell cycle
  • DNA-binding
  • sleeping sickness;
  • Trypanosoma brucei

Cite this

Thomas, James ; Baker, Nicola ; Dominicus, Caia ; Trenaman, Anna ; Glover, Lucy ; Alsford, Sam ; Horn, David. / Insights into antitrypanosomal drug mode-of-action from cytology-based profiling. In: PLoS Neglected Tropical Diseases. 2018 ; Vol. 12, No. 11. pp. 1-19.
@article{97b6012e83734d56b366252af92bdf09,
title = "Insights into antitrypanosomal drug mode-of-action from cytology-based profiling",
abstract = "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.",
keywords = "Cell cycle, DNA-binding, sleeping sickness;, Trypanosoma brucei",
author = "James Thomas and Nicola Baker and Caia Dominicus and Anna Trenaman and Lucy Glover and Sam Alsford and David Horn",
note = "Wellcome grant (100320/Z/12/Z).",
year = "2018",
month = "11",
day = "26",
doi = "10.1371/journal.pntd.0006980",
language = "English",
volume = "12",
pages = "1--19",
journal = "PLoS Neglected Tropical Diseases",
issn = "1935-2727",
publisher = "Public Library of Science",
number = "11",

}

Insights into antitrypanosomal drug mode-of-action from cytology-based profiling. / Thomas, James ; Baker, Nicola; Dominicus, Caia ; Trenaman, Anna; Glover, Lucy; Alsford, Sam; Horn, David (Lead / Corresponding author).

In: PLoS Neglected Tropical Diseases, Vol. 12, No. 11, e0006980, 26.11.2018, p. 1-19.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Insights into antitrypanosomal drug mode-of-action from cytology-based profiling

AU - Thomas, James

AU - Baker, Nicola

AU - Dominicus, Caia

AU - Trenaman, Anna

AU - Glover, Lucy

AU - Alsford, Sam

AU - Horn, David

N1 - Wellcome grant (100320/Z/12/Z).

PY - 2018/11/26

Y1 - 2018/11/26

N2 - 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.

AB - 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.

KW - Cell cycle

KW - DNA-binding

KW - sleeping sickness;

KW - Trypanosoma brucei

UR - http://www.scopus.com/inward/record.url?scp=85058012653&partnerID=8YFLogxK

U2 - 10.1371/journal.pntd.0006980

DO - 10.1371/journal.pntd.0006980

M3 - Article

VL - 12

SP - 1

EP - 19

JO - PLoS Neglected Tropical Diseases

JF - PLoS Neglected Tropical Diseases

SN - 1935-2727

IS - 11

M1 - e0006980

ER -