Polymorphisms in the K13-Propeller Gene in Artemisinin-Resistant Plasmodium in Mice

Shaoqin Zheng; Yuan Liang; Zhaojia Wang; Min Liu; Yingyi Chen; Ying Ai; Wenfeng Guo; Guoming Li; Yueming Yuan; Zhiyong Xu; Wanting Wu; Xinan Huang; Zhibing Wu; Qin Xu; Jianping Song; Changsheng Deng

doi:10.2147/IDR.S383127

Polymorphisms in the K13-Propeller Gene in Artemisinin-Resistant Plasmodium in Mice

Infect Drug Resist. 2022 Nov 8:15:6533-6544. doi: 10.2147/IDR.S383127. eCollection 2022.

Authors

Shaoqin Zheng^#^{1

2}, Yuan Liang^#², Zhaojia Wang¹, Min Liu¹, Yingyi Chen¹, Ying Ai¹, Wenfeng Guo^{1

3}, Guoming Li¹, Yueming Yuan^{1

2}, Zhiyong Xu², Wanting Wu¹, Xinan Huang^{1

3}, Zhibing Wu³, Qin Xu^{1

3}, Jianping Song^{1

3}, Changsheng Deng^{1

3}

Affiliations

¹ Artemisinin Research Center, Guangzhou University of Chinese Medicine, Guangzhou, People's Republic of China.
² Science and Technology Institute, Guangzhou University of Chinese Medicine, Guangzhou, People's Republic of China.
³ The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, People's Republic of China.

^# Contributed equally.

Abstract

Introduction: Artemisinin-based combination therapies (ACTs) act as first-line antimalarial drugs and play a crucial role in the successful control of falciparum malaria. However, the recent emergence of resistance of Plasmodium falciparum to ACTs in South East Asia is of particular concern. Hence, there is an urgent need to identify the genetic determinants of and understand the molecular mechanisms underpinning such resistance. Artemisinin resistance (AR) is primarily driven by the mutations in the P. falciparum K13 protein, which is widely recognized as the major molecular marker of AR. However, association of K13 mutations with in vivo AR has been ambiguous due to the absence of a tractable model.

Methods: In this study, we have successfully produced artemisinin- and piperaquine-resistant P. berghei K173 following drug administrations. Prolonged parasite clearance and early recrudescence were found following daily exposure to high doses of artemisinin and piperaquine. We have also sequenced the DNA of artemisinin-resistant strains and piperaquine-resistant strains of P. berghei K173 to explore the relationship between PfK13 and AR.

Results: The resistance index of P. berghei K173 reached 12.4 after 30 artemisinin-resistant generations, but AR declined gradually after 30 generations. On the 50th generation, the resistance index of artemisinin-resistant strains was only 5.0 compared with the severe drug resistance of piperaquine-resistant strains (I₉₀=148.8). DNA sequencing of artemisinin-resistant strains showed that there were 9 meaningful mutations at P. berghei K13-propeller domain, but the above mutations did not include common clinical point mutations.

Conclusion: Our data show that artemisinin is less susceptible to severe resistance compared with other antimalarial drugs. In addition, mutation on P. berghei K13 has a multi-drug-resistant phenotype and may be used as a biomarker to monitor its resistance. More studies need to be conducted on the new mutations detected so as to understand their association, if any, with ACT resistance.

Keywords: P. berghei K13 gene; Plasmodium berghei; artemisinin resistance; malaria.

Grants and funding

This work was supported by grants from Natural Science Foundation of China [Grant Number 82074301,81403295 and 81873218] to CD and SJ, and Project of Traditional Chinese Medicine Bureau of Guangdong [Grant Number 20201084] to SZ. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.