Apicoplast ribosomal protein S10-V127M enhances artemisinin resistance of a Kelch13 transgenic Plasmodium falciparum

Abstract

Background The resistance of Plasmodium falciparum to artemisinin-based (ART) drugs, the front-line drug family used in artemisinin-based combination therapy (ACT) for treatment of malaria, is of great concern. Mutations in the kelch13 (k13) gene (for example, those resulting in the Cys580Tyr [C580Y] variant) were identified as genetic markers for ART-resistant parasites, which suggests they are associated with resistance mechanisms. However, not all resistant parasites contain a k13 mutation, and clearly greater understanding of resistance mechanisms is required. A genome-wide association study (GWAS) found single nucleotide polymorphisms associated with ART-resistance in fd (ferredoxin), arps10 (apicoplast ribosomal protein S10), mdr2 (multidrug resistance protein 2), and crt (chloroquine resistance transporter), in addition to k13 gene mutations, suggesting that these alleles contribute to the resistance phenotype. The importance of the FD and ARPS10 variants in ART resistance was then studied since both proteins likely function in the apicoplast, which is a location distinct from that of K13. Methods The reported mutations were introduced, together with a mutation to produce the k13-C580Y variant into the ART-sensitive 3D7 parasite line and the effect on ART-susceptibility using the 0−3 h ring survival assay (RSA0−3 h) was investigated. Results and conclusion Introducing both fd-D193Y and arps10-V127M into a k13-C580Y-containing parasite, but not a wild-type k13 parasite, increased survival of the parasite in the RSA0−3 h. The results suggest epistasis of arps10 and k13, with arps10-V127M a modifier of ART susceptibility in different k13 allele backgrounds.

Ribosome Specialization in Protozoa Parasites

“…Likewise, MRPL12/bL12m, MRPL17/bL17m, MRPL18/uL18m, and MRPL23/ul23m are important for proper enzymatic activity of cytochrome bc 1 complex and the resistance to bc 1 inhibitors [ 91 , 96 ]. Additionally, the protein ARPS10 from the apicoplast has shown an important role in tolerance to febrile temperatures and artemisinin, showing the role of this plastids in drug resistance [ 97 , 98 ].…”

Antagonistic antimalarial properties of a methoxyamino chalcone derivative and 3-hydroxypyridinones in combination with dihydroartemisinin againstPlasmodium falciparum

“…The 3D7 parasite line and the transgenic 3D7 parasite lines in this study, which were established in our earlier study, were cultured and maintained in the same conditions in which the parasites grew at the same rate in these culture conditions [27]. Briefly, the parasites were cultured in complete RPMI 1640 medium pH7.4 containing 2 mM L-glutamine, 25 mM HEPES, 2 g/L NaHCO 3 , 27.2 mg/L hypoxanthine, and 0.5% Albumax II using human O + blood group erythrocytes (2-4% hematocrit).…”

“…Therefore, it is unclear whether the parasite genetic background, in particular, the allelic status of the fd and k13 genes affects the antimalarial potency of compounds targeting Fd/FNR and/or the interaction of these compounds with DHA. Currently, we have identified growth inhibition in the ART−sensitive P. falciparum 3D7 strain and genome−edited lines of this parental strain with fd and k13 mutations (fd D193Y _3D7, k13 C580Y _3D7, and k13 C580Y fd D193Y _3D7 [27]. We hypothesized that the antimalarial potency of chalcone derivatives C3 and our iron chelators and their interactions with DHA would be affected by the genetic background of the parasite.…”

“…For example, iron chelators are antagonistic to ART and related compounds [14,23], which is explained by the iron-dependent activation of ART [21,24,25]. Although the putative loss of function in the fd -D193Y mutant is associated with ART resistance, it does not affect the survival of P. falciparum parasites exposed to DHA [26,27]. Therefore, it is unclear whether the parasite genetic background, in particular, the allelic status of the fd and k13 genes affects the antimalarial potency of compounds targeting Fd/FNR and/or the interaction of these compounds with DHA.…”

Antagonistic antimalarial properties of a methoxyamino chalcone derivative and 3-hydroxypyridinones in combination with dihydroartemisinin against Plasmodium falciparum

“…Iron chelators are antagonistic to ART and related compounds ( Antoine et al, 2014 ; Suwito et al, 2014 ), which can be explained by the iron-dependent activation of ART ( Meshnick et al, 1993 ; Pattanapanyasat et al, 2001 ; Xie et al, 2016 ; Klonis, Creek & Tilley, 2013 ). Although the putative loss of function in the fd -D193Y mutant is associated with ART resistance, it does not affect the survival of P. falciparum parasites exposed to DHA ( Kampoun et al, 2022 ; Stokes et al, 2021 ). Therefore, it is unclear whether the parasite genetic background, in particular, the allelic status of the fd and k13 genes, affects the antimalarial potency of compounds targeting Fd/FNR and/or the interaction of these compounds with DHA.…”