Pan-active imidazolopiperazine antimalarials target the Plasmodium falciparum intracellular secretory pathway

A promising new compound class for treating human malaria is the imidazolopiperazines (IZP) class. IZP compounds KAF156 (Ganaplacide) and GNF179 are effective against Plasmodium symptomatic asexual blood-stage infections, and are able to prevent transmission and block infection in animal models. But despite the identification of resistance mechanisms in P. falciparum, the mode of action of IZPs remains unknown. To investigate, we here combine in vitro evolution and genome analysis in Saccharomyces cerevisiae with molecular, metabolomic, and chemogenomic methods in P. falciparum. Our findings reveal that IZP-resistant S. cerevisiae clones carry mutations in genes involved in Endoplasmic Reticulum (ER)-based lipid homeostasis and autophagy. In Plasmodium, IZPs inhibit protein trafficking, block the establishment of new permeation pathways, and cause ER expansion. Our data highlight a mechanism for blocking parasite development that is distinct from those of standard compounds used to treat malaria, and demonstrate the potential of IZPs for studying ER-dependent protein processing.

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Elizabeth Winzeler
Feb 1, 2020 No software was used to collect data in this study Yeast Genome Sequencing DNA sequencing was done using Illumina HiSeq 2500 in RapidRun mode to generate paired-end reads 100bp in length. Resulting reads were then aligned to the S. cerevisiae 288C reference genome (assembly R64) using BWA-mem and further processed using Picard Tools (http://broadinstitute.github.io/picard/). SNVs and INDELs were called using GATK HaplotypeCaller, filtered based on GATK recommendations and annotated with SnpEff.
Yeast Functional Genetics Analysis Gene maps were generated using Illustrator of Biological Sequences (IBS) software package, while Protein-Protein Interaction (PPI) networks were generated using the STRING database.
Evaluation of Drug Sensitivity (IC50) GraphPad Prism (commercially available software) was used to plot dose-response curves and calculate IC50s for all datasets Sphingolipid Analysis Sphingolipid quantitation was performed using Agilent Quantitative Analysis Software and compared using GraphPad Prism nature research | reporting summary

October 2018
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