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Comparative transcript expression analysis of miltefosine-sensitive and miltefosine-resistant Leishmania donovani

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Abstract

Leishmania donovani is the causative agent of anthroponotic visceral leishmaniasis in the Indian subcontinent. Oral miltefosine therapy has recently replaced antimonials in endemic areas. However, the drug is at risk of emergence of resistance due to unrestricted use, and, already, there are indications towards decline in treatment efficacy. Hence, understanding the mechanism of miltefosine resistance in the parasite is crucial. We employed genomic microarray analysis to compare the gene expression patterns of miltefosine-resistant and miltefosine-sensitive L. donovani. Three hundred eleven genes, representing ∼3.9 % of the total Leishmania genome, belonging to various functional categories including metabolic pathways, transporters, and cellular components, were differentially expressed in miltefosine-resistant parasite. Results in the present study highlighted the probable mechanisms by which the parasite sustains miltefosine pressure including (1) compromised DNA replication/repair mechanism, (2) reduced protein synthesis and degradation, (3) altered energy utilization via increased lipid degradation, (4) increased ABC 1-mediated drug efflux, and (5) increased antioxidant defense mechanism via elevated trypanothione metabolism. The study provided the comprehensive insight into the underlying mechanism of miltefosine resistance in L. donovani that may be useful to design strategies to increase lifespan of this important oral antileishmanial drug.

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Acknowledgments

We are thankful to Dr Marc Ouellette at the Research Centre in Infectious Diseases, Faculty of Medicine, Laval University, Quebec, Canada for kindly sharing the microarray design with us. AK and VS are grateful to Council for Scientific and Industrial Research and University Grants Commission, India, respectively, for providing research fellowship. This work was supported by Indian Council of Medical Research grant number 63/4/2007/-BMS.

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The authors do not have commercial or other associations that might pose any competing interest.

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  1. Arpita Kulshrestha

    Present address: Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA

Authors and Affiliations

  1. National Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi, 110029, India

    Arpita Kulshrestha, Vanila Sharma, Ruchi Singh & Poonam Salotra

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  1. Arpita Kulshrestha
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  2. Vanila Sharma
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Correspondence to Poonam Salotra.

Electronic supplementary material

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S1.1

Primers used to amplify LdMT and LdRos3 full-length gene sequence (DOCX 11 kb)

S1.2

Primers used to sequence LdMT and LdRos3 from both sensitive and resistant cell lines (DOC 36 kb)

S2

Primers used for quantitative real-time RT-PCR expression analysis. This table contains the sequences of the primers used for quantitative real-time PCR expression analysis to validate DNA microarray studies (DOCX 15 kb)

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Kulshrestha, A., Sharma, V., Singh, R. et al. Comparative transcript expression analysis of miltefosine-sensitive and miltefosine-resistant Leishmania donovani . Parasitol Res 113, 1171–1184 (2014). https://doi.org/10.1007/s00436-014-3755-6

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