Within-host selection of drug resistance in a mouse model of repeated interrupted treatment of Plasmodium yoelii infection

Suci Nuralitha,Andy I M Hoepelman,Josephine E Siregar,Sangkot Marzuki,Din Syafruddin

doi:10.1186/s12936-017-1860-6

Suci Nuralitha, Andy I M Hoepelman + Show 3 more

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https://doi.org/10.1186/s12936-017-1860-6

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Abstract

BackgroundTo study within-host selection of resistant parasites, an important factor in the development of resistance to anti-malarial drugs, a mouse model of repeated interrupted malaria treatment (RIT) has been developed. The characteristics of within host selection of resistance to atovaquone and pyrimethamine in Plasmodium yoelii was examined in such a model.MethodsTreatment of P. yoelii infected mice, with atovaquone or pyrimethamine, was started at parasitaemia level of 3–5%, interrupted when reduced to less than 0.4%, and restarted following parasitaemia recovery to the initial level. Treatment cycles were repeated until stable phenotype resistance was observed.ResultsPlasmodium yoelii rapidly developed resistance to atovaquone (2.75 ± 1.06 cycles) and to pyrimethamine (5.4 ± 0.89 cycles) under RIT. A dose dependent phenomenon in the selection of atovaquone resistance mutations was observed. All mutations that underlie resistance to therapeutic doses of 0.3–1.44 mg kg−1 BW were found to be in the Qo2 domain of the cytochrome b gene (I258M, F267I/L/S, L271V, K272R, L271V and K272R). Those associated with lower doses of 0.01–0.03 mg kg−1 BW were in the Qo1 domain (M133I and T139S). The resistance mutations occurred at four of the 16 atovaquone putative drug binding sites suggested in P. falciparum.ConclusionsRIT of P. yoelii infected mice led to rapid development of resistance to atovaquone and pyrimethamine. The dose dependent selection of resistance mutants to atovaquone observed during RIT might reflect the outcome of two different causes of malaria treatment failure in human, repeated incomplete treatment with therapeutic dose and repeated inadequate treatment associated with sub-therapeutic dose, and need to be systematically investigated.

Highlights

To study within-host selection of resistant parasites, an important factor in the development of resistance to anti-malarial drugs, a mouse model of repeated interrupted malaria treatment (RIT) has been developed
It is important to expand the study to examine the development of resistance over a wider range of atovaquone doses, as resistance resulting from repeated incomplete treatment with therapeutic dose in P. berghei is associated with mutations mostly in the quinone binding 2 (Qo2) domain of the cytb gene (Y268C/N/S, L271V and K272R) [1]; in contrast, isolation of atovaquone resistant mutants of P. berghei and Plasmodium yoelii by serial technique with increasing doses of the drug (ST) [4, 5], resulted in resistant parasites with underlying mutations in or near the quinone binding 1 (Qo1) domain (M133I, L144S, V284F)
Rapid development of P. yoelii resistance to atovaquone and pyrimethamine during repeated interrupted malaria treatment (RIT) The typical RIT experiments shown in Fig. 1, illustrating the cycles of interrupted treatment of P. yoelii-infected mice with atovaquone or pyrimethamine, demonstrates that stable resistant phenotype was already established in the third treatment cycle with atovaquone (Fig. 1a), compare to five treatment cycles for pyrimethamine (Fig. 1b)

Summary

Introduction

To study within-host selection of resistant parasites, an important factor in the development of resistance to anti-malarial drugs, a mouse model of repeated interrupted malaria treatment (RIT) has been developed. The characteristics of within host selection of resistance to atovaquone and pyrimethamine in Plasmodium yoelii was examined in such a model. It is important to expand the study to examine the development of resistance over a wider range of atovaquone doses, as resistance resulting from repeated incomplete treatment with therapeutic dose in P. berghei is associated with mutations mostly in the quinone binding 2 (Qo2) domain of the cytb gene (Y268C/N/S, L271V and K272R) [1]; in contrast, isolation of atovaquone resistant mutants of P. berghei and Plasmodium yoelii by serial technique with increasing doses of the drug (ST) [4, 5], resulted in resistant parasites with underlying mutations in or near the quinone binding 1 (Qo1) domain (M133I, L144S, V284F)

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