Mechanism of artemisinin resistance in selected malaria parasite lines

描述（由申请人提供）：耐多药疟疾寄生虫恶性疟原虫是导致全球疟疾死灰复燃的主要因素。由于许多抗疟药物失效，大多数疟疾流行国家采用了以青蒿素为基础的联合疗法来治疗恶性疟疾。虽然这两类抗疟药是唯一没有证实临床耐药的抗疟药，但世界许多地区关于青蒿素治疗失败的报告令人极为关切。人们对青蒿素的确切作用方式和耐药性机制知之甚少，这使得在部署青蒿素的地区进行耐药性监测非常困难。在此，我们建议使用实验室选择的青蒿素耐药寄生虫克隆来阐明青蒿素耐药机制。利用最近开发的全基因组分析工具，我们建议1)利用高密度平铺阵列鉴定与抗性寄生虫相关的单核苷酸多态性（snp）和拷贝数变异（CNVs）；2)利用全基因组表达芯片和定量蛋白质组学技术筛选出可能导致青蒿素耐药的序列多态性。这些高通量方法将允许交叉检查和鉴定候选多态性，可以在未来的研究中验证。

DESCRIPTION (provided by applicant): Multidrug-resistant malaria parasite Plasmodium falciparum is a major contributing factor to the global resurgence of malaria. With the failing of many antimalarial drugs, most malaria- endemic countries have adopted artemisinin-based combination therapy (ACT) to treat falciparum malaria. Although these are the only class of antimalarial drugs to which there is no confirmed clinical resistance, the reports of artemisinin treatment failures in many regions of the world are of great concern. The exact mode of action of artemisinins and the mechanism of resistance are poorly understood, making resistance surveillance in areas of artemisinin deployment very difficult. Here, we propose to use laboratory selected artemisinin-resistant parasite clones for elucidating the mechanism of artemisinin resistance. Using recently developed genome-wide analysis tools, we propose to 1) identify single nucleotide polymorphisms (SNPs) and copy number variations (CNVs) associated with the resistant parasites using high-density tiling arrays; and 2) filter out the sequence polymorphisms that might be responsible for the observed resistance to artemisinin using genome-wide expression microarray and quantitative proteomics. These high-throughput approaches will allow cross examination and identification of candidate polymorphisms that can be validated in future studies. PUBLIC HEALTH RELEVANCE: In response to the escalating problem of drug resistance in malaria control, artemisinin-based combinatory therapy (ACT) is being adopted by many malaria-endemic nations. This study aims at better understanding the resistance mechanism using laboratory selected artemisinin-resistant lines. The results from this study may allow us to design and implement countermeasures to closely monitor and prevent resistance development to artemisinins in field parasite populations.