CYP6 P450 enzymes and ACE-1 duplication produce extreme and multiple insecticide resistance in the malaria mosquito Anopheles gambiae.
CYP6 P450 enzymes and ACE-1 duplication produce extreme and multiple insecticide resistance in the malaria mosquito Anopheles gambiae.
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DOI：
10.1371/journal.pgen.1004236
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发表时间：
2014-03
期刊：
影响因子：
4.5
通讯作者：
Weetman D
中科院分区：
文献类型：
--
作者：
Edi CV;Djogbénou L;Jenkins AM;Regna K;Muskavitch MA;Poupardin R;Jones CM;Essandoh J;Kétoh GK;Paine MJ;Koudou BG;Donnelly MJ;Ranson H;Weetman D
疟疾防治严重依赖拟除虫菊酯类杀虫剂，而按蚊对拟除虫菊酯类杀虫剂的敏感性正在下降。为了消除拟除虫菊酯抗药性，提倡使用替代杀虫剂进行室内滞留喷洒，氨基甲酸酯越来越重要。因此，西非科特迪瓦Tiassalé的冈比亚按蚊出现了非常强的氨基甲酸酯耐药性表型，这是一个潜在的重大业务挑战，特别是因为这些疟疾病媒现在对多种杀虫剂类别表现出耐药性。我们调查了安对最常用的氨基甲酸酯，苯虫威，抗性的遗传基础。来自Tiassalé的冈比亚。地理复制的全基因组微阵列实验确定升高的P450酶表达与苯虫威抗性相关，最显著的是来自CYP 6亚家族的基因。P450进一步涉及抗性表型诱导显着升高死亡率bendiocarb的胡椒基丁醚（PBO），这也增强了拟除虫菊酯和有机磷酸酯的作用。CYP 6P 3，特别是CYP 6 M2通过转基因表达在果蝇中产生了苯虫威抗性，除了这两个基因的拟除虫菊酯抗性和CYP 6 M2表达的DDT抗性之外。因此，CYP 6 M2可以引起对三种不同类别的杀虫剂的抗性，尽管氨基甲酸酯的生化机制尚不清楚，因为与CYP 6P 3相反，重组CYP 6 M2在体外不代谢恶虫威。强抗虫威蚊子也表现出乙酰胆碱酯酶ACE-1基因的表达升高，至少部分来自基因复制，这赋予抗性ACE-1 G119 S等位基因的额外拷贝的携带者生存优势。我们的结果对于基于病媒的疟疾控制来说是令人震惊的。Tiassalé An的极端氨基甲酸酯抗性。gambiae是由过度表达的靶位点等位基因变体与升高的CYP 6 P450表达偶联引起的，这也提供了对对比杀虫剂的抗性。表现出极端和交叉抗性的苔藓虫种群可能对标准的杀虫剂抗性管理做法反应迟钝。疟疾的控制主要依赖于按蚊对四类杀虫剂的抗药性越来越强。重要的是要仔细管理杀虫剂的使用，以尽量减少抗药性的增加，例如通过组合或轮换使用不同的化合物。最近，在西非的Tiassalé发现了对所有可用杀虫剂具有抗药性的蚊子，这可能会给抗药性管理带来问题，特别是如果共同的遗传机制是原因（“交叉抗药性”）。Tiassalé蚊子还对两种最重要的杀虫剂-拟除虫菊酯和氨基甲酸酯-表现出极强的抗药性。我们调查了Tiassalé极端氨基甲酸酯耐药性和交叉耐药性的遗传基础，以及结果在多哥其他人群中的适用性。我们发现，特定的P450酶参与了极端和交叉抗性，包括CYP 6 M2，它可以导致对三种杀虫剂的抗性。然而，编码乙酰胆碱酯酶（氨基甲酸酯和有机磷杀虫剂的靶位点）的基因的突变版本的扩增也起着重要作用。涉及极端耐药性和交叉耐药性的机制很可能对杀虫剂管理做法具有很强的适应性，这对以蚊子为目标的疟疾控制来说是一种令人担忧的情况。
Malaria control relies heavily on pyrethroid insecticides, to which susceptibility is declining in Anopheles mosquitoes. To combat pyrethroid resistance, application of alternative insecticides is advocated for indoor residual spraying (IRS), and carbamates are increasingly important. Emergence of a very strong carbamate resistance phenotype in Anopheles gambiae from Tiassalé, Côte d'Ivoire, West Africa, is therefore a potentially major operational challenge, particularly because these malaria vectors now exhibit resistance to multiple insecticide classes. We investigated the genetic basis of resistance to the most commonly-applied carbamate, bendiocarb, in An. gambiae from Tiassalé. Geographically-replicated whole genome microarray experiments identified elevated P450 enzyme expression as associated with bendiocarb resistance, most notably genes from the CYP6 subfamily. P450s were further implicated in resistance phenotypes by induction of significantly elevated mortality to bendiocarb by the synergist piperonyl butoxide (PBO), which also enhanced the action of pyrethroids and an organophosphate. CYP6P3 and especially CYP6M2 produced bendiocarb resistance via transgenic expression in Drosophila in addition to pyrethroid resistance for both genes, and DDT resistance for CYP6M2 expression. CYP6M2 can thus cause resistance to three distinct classes of insecticide although the biochemical mechanism for carbamates is unclear because, in contrast to CYP6P3, recombinant CYP6M2 did not metabolise bendiocarb in vitro. Strongly bendiocarb resistant mosquitoes also displayed elevated expression of the acetylcholinesterase ACE-1 gene, arising at least in part from gene duplication, which confers a survival advantage to carriers of additional copies of resistant ACE-1 G119S alleles. Our results are alarming for vector-based malaria control. Extreme carbamate resistance in Tiassalé An. gambiae results from coupling of over-expressed target site allelic variants with heightened CYP6 P450 expression, which also provides resistance across contrasting insecticides. Mosquito populations displaying such a diverse basis of extreme and cross-resistance are likely to be unresponsive to standard insecticide resistance management practices. Malaria control depends heavily on only four classes of insecticide to which Anopheles mosquitoes are increasingly resistant. It is important to manage insecticide application carefully to minimise increases in resistance, for example by using different compounds in combination or rotation. Recently, mosquitoes resistant to all available insecticides have been found in Tiassalé, West Africa, which could be problematic for resistance management, particularly if common genetic mechanisms are responsible (‘cross-resistance’). Tiassalé mosquitoes also exhibit extreme levels of resistance to the two most important classes, pyrethroids and carbamates. We investigated the genetic basis of extreme carbamate resistance and cross-resistance in Tiassalé, and the applicability of results in an additional population from Togo. We find that specific P450 enzymes are involved in both extreme and cross-resistance, including one, CYP6M2, which can cause resistance to three insecticide classes. However, amplification of a mutated version of the gene which codes for acetycholinesterase, the target site of both the carbamate and organophosphate insecticides, also plays an important role. Mechanisms involved in both extreme resistance and cross resistance are likely to be very resilient to insecticide management practices, and represent an alarming scenario for mosquito-targeted malaria control.
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影响因子：
3
作者：
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