Pyrethroid resistance in the New World malaria vector Anopheles albimanus is mediated by cytochrome P450 CYP6P5

Kusimo, Michael O.; Mackenzie-Impoinvil, Lucy; Ibrahim, Sulaiman; Muhammad, Abdullahi; Irving, Helen; Hearn, Jack; Lenhart, Audrey; Wondji, Charles S.

doi:10.1016/j.pestbp.2022.105061

Cited by 8 publications

(3 citation statements)

References 37 publications

(56 reference statements)

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“…To confirm whether the overexpression of CYP6P3-205D can confer cross resistance to pyrethroid and other insecticide classes, transgenic flies overexpressing the CYP6P3 were generated using the GAL4-UAS system following the protocols previously described (23, 24). The CYP6P3-205D allele was amplified using Phusion Taq polymerase with the allele specific primers (Table S1) and ligated to the D. melanogaster expression vector pUASattB pre-digested with BglII and KpnI restriction enzymes (New England Biolabs, Hitchin, UK).…”

Section: Suppl Materials and Methodsmentioning

confidence: 99%

A single E205D allele of a key P450CYP6P3is driving metabolic pyrethroid resistance in the major African malaria vectorAnopheles gambiae

Kengne-Ouafo,

Kouamo,

Muhammad

et al. 2024

Preprint

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Deciphering the molecular drivers of insecticide resistance is paramount to extend the effectiveness of malaria vector control tools. Here, we demonstrated that the E205D amino acid change in a key metabolic resistance P450CYP6P3drives pyrethroid resistance in the major malaria vector,Anopheles gambiae. Spatio-temporal whole genome Poolseq analyses in Cameroon detected a major P450-linked locus on chromosome 2R beside the sodium channel locus.In vitrometabolism assays with recombinantly expressedCYP6P3protein revealed that the catalytic efficiency of 205D was 2.5 times higher than E205 with α-cypermethrin. Similar patterns were observed for permethrin. Overexpression of the 205D allele in transgenic flies confers higher more pyrethroids and carbamates resistance, compared to controls. A DNA-based assay further supported that theCYP6P3-205D variant strongly correlates with pyrethroid resistance in field populations (OR=26.4; P<0.0001) and that it reduces the efficacy of pyrethroid-only LLINs with homozygote RR genotype exhibiting significantly higher survival following PermaNet 3.0 exposure compared to the SS genotype (OR: 6.1, p = 0.0113). Furthermore, theCYP6P3-E205D combines with thekdrtarget-site resistance mechanisms to worsen the loss of bednet efficacy. The 205D mutation is now predominant in West and Central Africa but less abundant or absent in East and South Africa with signs of introgression withAn. coluzziiin Ghana. This study highlights the importance of P450-based resistance and designs field-applicable tools to easily track the spread of metabolic resistance and assess its impact on control interventions.One Sentence Summary:The major obstacle to malaria control and elimination is the spread of parasite resistance to anti-malarial drugs, and mosquito resistance to insecticides. In this study, we identified a key point mutation E205D in the metabolic geneCYP6P3(cytochrome P450) conferring resistance to pyrethroids by enhancing the breakdown of insecticides used for bednets impregnation. DNA-based assays were then designed and used to determine the spread of the resistance across Africa and demonstrate that theCYP6P3-205D allele works together with the knockdown resistance in the voltage-gated Sodium channel to reduce the efficacy of insecticide-treated bednets.

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Section: Suppl Materials and Methodsmentioning

confidence: 99%

A single E205D allele of a key P450CYP6P3is driving metabolic pyrethroid resistance in the major African malaria vectorAnopheles gambiae

Kengne-Ouafo,

Kouamo,

Muhammad

et al. 2024

Preprint

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“…The CYP6P4a mutant allele has greater binding affinity for pyrethroids. In silico modelling and docking has been an essential tool in the characterisation of enzyme-substrate interactions of many P450s [34][35][36] . The hydroxylation or hydrolysis of the 4'-phenoxybenzyl moiety along with the cis/trans methyl spot of type I (permethrin) 37 and type II (deltamethrin) 25,38 pyrethroids are reported to be the main routes for pyrethroid metabolism.…”

Section: Allelic Variation Enhances Cyp6p4a and Cyp6p4b Pyrethroid De...mentioning

confidence: 99%

Two highly selected mutations in the tandemly duplicatedCYP6P4aandCYP6P4bdrive pyrethroid resistance inAnopheles funestus

Tatchou-Nebangwa,

Mugenzi,

Muhammad

et al. 2024

Preprint

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Gaining a comprehensive understanding of the genetic mechanisms underlying insecticide resistance in malaria vectors is crucial for optimising the effectiveness of insecticide-based vector control methods and developing diagnostic tools for resistance management. Considering the heterogeneity of metabolic resistance in major malaria vectors, the implementation of tailored resistance management strategies is essential for successful vector control. In this study, we provide evidence demonstrating that two highly selected mutations in the tandemly duplicated cytochrome P450 genes namely CYP6P4a and CYP6P4b, are driving pyrethroid insecticide resistance in the major malaria vector Anopheles funestus, in West Africa. Through a continent-wide polymorphism survey, we observed heightened indications of directional selection in both genes between 2014 and 2021. By conducting in vitro insecticide metabolism assays with recombinant enzymes expressed from both genes, we established that mutant alleles under selection exhibit higher metabolic efficiency compared to their wild-type counterparts. Furthermore, using the GAL4-UAS transgenic system, we demonstrated that transgenic Drosophila melanogaster flies overexpressing mutant alleles displayed an increased resistance to pyrethroids. These findings were in agreement with in silico characterisation, which highlighted changes in enzyme active site architecture that enhance the affinity of mutant alleles for type I and II pyrethroids. Furthermore, we developed two DNA-based assays capable of detecting the CYP6P4a-M220I and CYP6P4b-D284E mutations, showing their current confinement to West Africa. Genotype/phenotype correlation analyses revealed that these markers are strongly associated with resistance to types I and II pyrethroids and combine to drastically reduce the efficacy of pyrethroid bednets. Overall, our study makes available two field-applicable insecticide resistance molecular markers that will help in the monitoring and better management of insecticide resistance in West Africa.

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“…Insecticide-treated nets (ITNs) and indoor residual spraying (IRS) are key components of malaria control strategies that have been effective in reducing malaria transmission, particularly between the years 2000 and 2015 [5] and are now threatened by insecticide resistance, especially to pyrethroids. Target site mutations [6-9] over-expression of metabolic enzymes [10][11][12][13][14][15][16][17], cuticular thickening [18,19] and changes in microbial compositions [20,21] have been described as mechanisms involved in conferring insecticide resistant phenotypes. Often, these studies examine these mechanisms as being responsible for the observed insecticide resistance individually, but this is rarely the case in that these mechanisms often collaborate and work in conjunction to bring about resistance [22].…”

Section: Introductionmentioning

confidence: 99%

Key gene modules and hub genes associated with pyrethroid and organophosphate resistance in Anopheles mosquitoes: A systems biology approach

Odhiambo,

Derilus,

Impoinvil

et al. 2024

Preprint

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Indoor residual spraying (IRS) and insecticide-treated nets (ITNs) are the main methods used to control mosquito populations for malaria prevention. The efficacy of these strategies is threatened by the spread of insecticide resistance (IR), limiting the success of malaria control. Studies of the genetic evolution leading to insecticide resistance could enable the identification of molecular markers that can be used for IR surveillance and an improved understanding of the molecular mechanisms associated with IR. This study used a weighted gene co-expression network analysis (WGCNA) algorithm, a systems biology approach, to identify genes with similar co-expression patterns (modules) and hub genes that are potential molecular markers for insecticide resistance surveillance in Kenya and Benin. A total of 20 and 26 gene co-expression modules were identified via the average linkage hierarchical clustering from Anopheles arabiensis and An. gambiae, respectively, and hub genes (highly connected genes) identified within each module. Four specific genes stood out: serine protease, E3 ubiquitin-protein ligase, cuticular protein RR2, and leucine-rich immune protein, which were top hub genes in both species and could serve as potential markers and targets for monitoring IR in these malaria vectors. In addition to the identified markers, we explored molecular mechanisms using enrichment maps that revealed a complex process involving multiple steps, from odorant binding and neuronal signaling to cellular responses, immune modulation, cellular metabolism, and gene regulation. Incorporation of these dynamics into the development of new insecticides and tracking of insecticide resistance could improve sustainable and cost-effective deployment of interventions.

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Pyrethroid resistance in the New World malaria vector Anopheles albimanus is mediated by cytochrome P450 CYP6P5

Cited by 8 publications

References 37 publications

A single E205D allele of a key P450CYP6P3is driving metabolic pyrethroid resistance in the major African malaria vectorAnopheles gambiae

A single E205D allele of a key P450CYP6P3is driving metabolic pyrethroid resistance in the major African malaria vectorAnopheles gambiae

Two highly selected mutations in the tandemly duplicatedCYP6P4aandCYP6P4bdrive pyrethroid resistance inAnopheles funestus

Key gene modules and hub genes associated with pyrethroid and organophosphate resistance in Anopheles mosquitoes: A systems biology approach

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