1H NMR-Based Metabolomic Analysis of Intraerythrocytic Stages of P. falciparum and Antimalarial Treatments

Abstract

1H NMR provides an indirect approach that can be used to study the metabolism of the malaria parasite, Plasmodium falciparum, in human erythrocytes. It allows for facile analysis of the metabolites that the parasite excretes into and consumes from the surrounding media to gain insight into this dynamic system. This study carefully processes a very large batch of NMR data using MestReNova through a rigorous procedure. This is followed by multivariate statistical analyses in SIMCA to metabolically understand how erythrocytes respond to parasitic infection and subsequent treatment with artemisinin, chloroquine, or a candidate antimalarial, tiamulin. Trajectories reveal separation between the infected and artemisinin sets, and the uninfected, chloroquine, and tiamulin sets. Coefficient plots provide an introduction to the specific metabolites, the concentrations of which change systematically in the media. This study conclusively demonstrates that tiamulin treatment results in highly similar metabolic effects to those of chloroquine, both in terms of overall time-evolution trajectories and metabolic exchange with the medium during each individual phase. It also appears to normalize cells to be similar to control uninfected cells. This cost effective and non-targeted 1H NMR approach to profile metabolites can be adopted for large-scale drug screenings.