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Biomarkers of antimony resistance: need for expression analysis of multiple genes to distinguish resistance phenotype in clinical isolates of Leishmania donovani

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Abstract

Resistance to antimony is a major cause of failure to therapy in a large proportion of visceral leishmaniasis cases. Methods to distinguish resistant and sensitive parasite are urgently needed as the standard in vitro intracellular drug susceptibility assays are cumbersome and time consuming. Differential expression profiling studies have led to the identification of several antimony resistance-associated genes; however, their efficacy as a potential biomarker for monitoring antimony resistance remains imprecise. We analysed the expression of eight genes [antimony metabolism-associated genes—multidrug resistance protein A (MRPA), γ-glutamylcysteine synthetase (γ-GCS) and aquaporin-1 (AQP1)—and genes identified by proteome/transcriptome profiling—heat shock protein 83, mitogen-activated protein kinase 1 and histones H1, H2A and H4) in antimony-resistant (n = 10) and antimony-sensitive (n = 4) clinical isolates of Leishmania donovani by quantitative real-time PCR, in comparison with a lab-generated resistant and a standard sensitive isolate. We observed a significant differential expression of MRPA, histone H1 (p < 0.01), γ-GCS, HSP83 (p < 0.005) and histone H2A and H4 (p < 0.0001) in a group of sodium antimony gluconate-resistant isolates compared to sensitive isolates. Preferential AQP1 expression was observed in all the sensitive isolates (p < 0.0001). Overall, expression profile in field isolates for all the genes studied showed altered expression in majority of isolates, while in some, the expression was static. All the isolates showed a mosaic of expression pattern of the genes analysed indicating constellation of genes contributes towards the drug susceptibility of parasite. As none of the genes exhibit an absolute correlation with phenotype, targeted expression analysis of a set of genes should be considered as biomarker for distinguishing the antimony-resistant and antimony-sensitive parasite.

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Abbreviations

AQP1:

Aquaporin-1

γ-GCS:

γ-Glutamylcysteine synthetase

HSP83:

Heat shock protein 83

MAPK1:

Mitogen-activated protein kinase 1

MRPA:

Multidrug resistance protein A

SAG:

Sodium antimony gluconate

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Acknowledgements

The authors are thankful to Jose M. Requena (Universidad Autonoma de Madrid, Spain) for anti-H2A antibodies. This work formed a part of PhD thesis of DK, submitted to BITS-Pilani, Rajasthan. Financial support of Indian Council of Medical Research, India is gratefully acknowledged. RS is a recipient of UNESCO L`oreal for Women in Science fellowship grant. DK and AK are grateful to the Council of Scientific and Industrial Research, India for fellowship.

Conflicts of interest

None to declare.

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Author notes

  1. Dhiraj Kumar

    Present address: Division of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, CBER, FDA, Kensington Bethesda, MD, USA

Authors and Affiliations

  1. National Institute of Pathology, Indian Council of Medical Research, Safdarjung Hospital Campus, New Delhi, India

    Dhiraj Kumar, Vasundhra Bhandari, Arpita Kulshrestha & Poonam Salotra

  2. National Institute of Malaria Research, New Delhi, India

    Ruchi Singh

  3. Department of Medicine, Safdarjung Hospital, New Delhi, 110029, India

    Narendra Singh Negi

Authors
  1. Dhiraj Kumar
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  2. Ruchi Singh
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  3. Vasundhra Bhandari
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  4. Arpita Kulshrestha
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  5. Narendra Singh Negi
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  6. Poonam Salotra
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Corresponding author

Correspondence to Poonam Salotra.

Additional information

Dhiraj Kumar and Ruchi Singh contributed equally to this work.

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Kumar, D., Singh, R., Bhandari, V. et al. Biomarkers of antimony resistance: need for expression analysis of multiple genes to distinguish resistance phenotype in clinical isolates of Leishmania donovani . Parasitol Res 111, 223–230 (2012). https://doi.org/10.1007/s00436-012-2823-z

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  • DOI: https://doi.org/10.1007/s00436-012-2823-z

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