A comparative transcriptomic analysis of replicating and dormant liver stages of the relapsing malaria parasite Plasmodium Cynomolgi

Abstract

Plasmodium liver hypnozoites, which cause disease relapse, are widely considered to be the last barrier towards malaria eradication. The biology of this quiescent form of the parasite is poorly understood which hinders drug discovery. We report a comparative transcriptomic dataset of replicating liver schizonts and dormant hypnozoites of the relapsing parasite Plasmodium cynomolgi. Hypnozoites express only 34% of Plasmodium physiological pathways, while 91% are expressed in replicating schizonts. Few known malaria drug targets are expressed in quiescent parasites, but pathways involved in microbial dormancy, maintenance of genome integrity and ATP homeostasis were robustly expressed. Several transcripts encoding heavy metal transporters were expressed in hypnozoites and the copper chelator neocuproine was cidal to all liver stage parasites. This transcriptomic dataset is a valuable resource for the discovery of vaccines and effective treatments to combat vivax malaria.

Data availability

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Annemarie Voorberg-van der Wel

    Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, Netherlands
    Competing interests
    No competing interests declared.
  2. Guglielmo Roma

    Chemical Biology and Therapeutics, Novartis Institutes for BioMedical Research, Basel, Switzerland
    Competing interests
    Guglielmo Roma, Employed by and/or shareholder of Novartis Pharma AG.
  3. Devendra Kumar Gupta

    Novartis Institute for Tropical Diseases, Novartis Institutes for BioMedical Research, Singapore, Singapore
    Competing interests
    Devendra Kumar Gupta, Employed by and/or shareholder of Novartis Pharma AG.
  4. Sven Schuierer

    Chemical Biology and Therapeutics, Novartis Institutes for BioMedical Research, Basel, Switzerland
    Competing interests
    Sven Schuierer, Employed by and/or shareholder of Novartis Pharma AG.
  5. Florian Nigsch

    Chemical Biology and Therapeutics, Novartis Institutes for BioMedical Research, Basel, Switzerland
    Competing interests
    Florian Nigsch, Employed by and/or shareholder of Novartis Pharma AG.
  6. Walter Carbone

    Developmental and Molecular Pathways, Novartis Institutes for BioMedical Research, Basel, Switzerland
    Competing interests
    Walter Carbone, Employed by and/or shareholder of Novartis Pharma AG.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6150-8295
  7. Anne-Marie Zeeman

    Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, Netherlands
    Competing interests
    No competing interests declared.
  8. Boon Heng Lee

    Novartis Institute for Tropical Diseases, Novartis Institutes for BioMedical Research, Singapore, Singapore
    Competing interests
    Boon Heng Lee, Employed by and/or shareholder of Novartis Pharma AG.
  9. Sam O Hofman

    Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, Netherlands
    Competing interests
    No competing interests declared.
  10. Bart W Faber

    Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, Netherlands
    Competing interests
    No competing interests declared.
  11. Judith Knehr

    Developmental and Molecular Pathways, Novartis Institutes for BioMedical Research, Basel, Switzerland
    Competing interests
    Judith Knehr, Employed by and/or shareholder of Novartis Pharma AG.
  12. Erica Pasini

    Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, Netherlands
    Competing interests
    No competing interests declared.
  13. Bernd Kinzel

    Chemical Biology and Therapeutics, Novartis Institutes for BioMedical Research, Basel, Switzerland
    Competing interests
    Bernd Kinzel, Employed by and/or shareholder of Novartis Pharma AG.
  14. Pablo Bifani

    Novartis Institute for Tropical Diseases, Novartis Institutes for BioMedical Research, Singapore, Singapore
    Competing interests
    Pablo Bifani, Employed by and/or shareholder of Novartis Pharma AG.
  15. Ghislain M C Bonamy

    Novartis Institute for Tropical Diseases, Novartis Institutes for BioMedical Research, Singapore, Singapore
    Competing interests
    Ghislain M C Bonamy, Employed by and/or shareholder of Novartis Pharma AG.
  16. Tewis Bouwmeester

    Chemical Biology and Therapeutics, Novartis Institutes for BioMedical Research, Basel, Switzerland
    Competing interests
    Tewis Bouwmeester, Employed by and/or shareholder of Novartis Pharma AG.
  17. Clemens H M Kocken

    Department of Parasitology, Biomedical Primate Research Centre, Rijswijk, Netherlands
    For correspondence
    kocken@bprc.nl
    Competing interests
    No competing interests declared.
  18. Thierry Tidiane Diagana

    Novartis Institute for Tropical Diseases, Novartis Institutes for BioMedical Research, Singapore, Singapore
    For correspondence
    thierry.diagana@novartis.com
    Competing interests
    Thierry Tidiane Diagana, Employed by and/or shareholder of Novartis Pharma AG.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8520-5683

Funding

Bill and Melinda Gates Foundation

  • Guglielmo Roma
  • Clemens H M Kocken
  • Thierry Tidiane Diagana

Medicines for Malaria Venture

  • Clemens H M Kocken
  • Thierry Tidiane Diagana

Wellcome

  • Clemens H M Kocken

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Urszula Krzych, Walter Reed Army Institute of Research, United States

Ethics

Animal experimentation: Ethics statementNonhuman primates were used because no other models (in vitro or in vivo) were suitable for the aims of this project. The local independent ethical committee constituted conform Dutch law (BPRC Dier Experimenten Commissie, DEC) approved the research protocol (agreement number DEC# 708) prior to the start and the experiments were all performed according to Dutch and European laws. The Council of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC International) has awarded BPRC full accreditation. Thus, BPRC is fully compliant with the international demands on animal studies and welfare as set forth by the European Council Directive 2010/63/EU, and Convention ETS 123, including the revised Appendix A as well as the 'Standard for humane care and use of Laboratory Animals by Foreign institutions' identification number A5539-01, provided by the Department of Health and Human Services of the United States of America's National Institutes of Health (NIH) and Dutch implementing legislation. The rhesus monkeys (Macaca mulatta, either gender, age 4-7 years, Indian or mixed origin) used in this study were captive-bred and socially housed. Animal housing was according to international guidelines for nonhuman primate care and use. Besides their standard feeding regime, and drinking water ad libitum via an automatic watering system, the animals followed an environmental enrichment program in which, next to permanent and rotating non-food enrichment, an item of food-enrichment was offered to the macaques daily. All animals were monitored daily for health and discomfort. All intravenous injections and large blood collections were performed under ketamine sedation, and all efforts were made to minimize suffering. Liver lobes were collected from monkeys that were euthanized in the course of unrelated studies (ethically approved by the BPRC DEC) or euthanized for medical reasons, as assessed by a veterinarian. Therefore, none of the animals from which liver lobes were derived were specifically used for this work, according to the 3Rrule thereby reducing the numbers of animals used. Euthanasia was performed under ketamine sedation (10 mg/kg) and was induced by intracardiac injection of euthasol 20%, containing pentobarbital.

Version history

  1. Received: June 15, 2017
  2. Accepted: December 5, 2017
  3. Accepted Manuscript published: December 7, 2017 (version 1)
  4. Version of Record published: January 8, 2018 (version 2)

Copyright

© 2017, Voorberg-van der Wel et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Annemarie Voorberg-van der Wel
  2. Guglielmo Roma
  3. Devendra Kumar Gupta
  4. Sven Schuierer
  5. Florian Nigsch
  6. Walter Carbone
  7. Anne-Marie Zeeman
  8. Boon Heng Lee
  9. Sam O Hofman
  10. Bart W Faber
  11. Judith Knehr
  12. Erica Pasini
  13. Bernd Kinzel
  14. Pablo Bifani
  15. Ghislain M C Bonamy
  16. Tewis Bouwmeester
  17. Clemens H M Kocken
  18. Thierry Tidiane Diagana
(2017)
A comparative transcriptomic analysis of replicating and dormant liver stages of the relapsing malaria parasite Plasmodium Cynomolgi
eLife 6:e29605.
https://doi.org/10.7554/eLife.29605

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https://doi.org/10.7554/eLife.29605

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    Relapses of Plasmodium dormant liver hypnozoites compromise malaria eradication efforts. New radical cure drugs are urgently needed, yet the vast gap in knowledge of hypnozoite biology impedes drug discovery. We previously unraveled the transcriptome of 6 to 7 day-old P. cynomolgi liver stages, highlighting pathways associated with hypnozoite dormancy (Voorberg-van der Wel et al., 2017). We now extend these findings by transcriptome profiling of 9 to 10 day-old liver stage parasites, thus revealing for the first time the maturation of the dormant stage over time. Although progression of dormancy leads to a 10-fold decrease in transcription and expression of only 840 genes, including genes associated with housekeeping functions, we show that pathways involved in quiescence, energy metabolism and maintenance of genome integrity remain the prevalent pathways active in mature hypnozoites.

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