Novel Genetic Mechanism of Artemisinin Resistance for Malaria

青蒿素抗疟疾的新遗传机制

• 批准号: 10201429
• 负责人: Daniel L HARTL
• 金额: $ 69.49万
• 依托单位: HARVARD SCHOOL OF PUBLIC HEALTH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-05 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201429
• 关键词: Africa; African; Alleles; Artemisinins; Asia; Asians; CRISPR/Cas technology; Cambodia; Cambodian; Child; Clinical; Collaborations; Combined Modality Therapy; Evolution; Exposure to; Family; Frequencies; Genes; Genetic; In Vitro; Incidence; Individual; Infection; Laboratories; Malaria; Measures; Molecular; Mutation; Other Genetics; Parasites; Phenotype; Population; Population Genetics; Proteins; Quality Control; Reporting; Resistance; Role; Sampling; Senegal; Sentinel; Site; Southeastern Asia; Structure; Tanzania; Testing; Validation; WD Repeat; base; biological adaptation to stress; coronin protein; deep sequencing; early detection biomarkers; gene interaction; genetic analysis; mutant; novel; novel therapeutics; pressure; prevent; resistance gene; resistant strain

Recent progress in malaria control has reduced the incidence and saved the lives of hundreds of thousands of children, but effective strategies depend on a combination of measures that includes special emphasis on artemisinin-based combination therapies (ACTs). The predicted dire consequences of the evolution and spread of artemisinin (ART) resistance have been borne out tragically in Southeast Asia, where ART resistance evolved quickly and spread rapidly. The ART-resistance determinants reside in the propeller domain of the Pfkelch13 locus (K13), which is thought to facilitate protein quality control and modulate stress responses. Importantly, the K13 determinant was first identified through five-year in vitro selection and sequencing of a resistant strain from Tanzania; K13 was only later confirmed in the field in Southeast Asia. Were ART resistance to take hold and spread in Africa it would be truly catastrophic. Why it has not is open to speculation, however one possibility is because K13 effects are strongly dependent on genetic background, and there are significant genetic differences between African parasite and SE Asian lineages. But persistent, strong selection pressure from ART treatment in Africa will—as every evolutionary biologist knows—result almost inevitably in the evolution of resistance determinants in Africa. Based on the hypothesis that ART resistance might depend on genetic background, four years ago (prior to the K13 report) we began selecting independent replicate lines of parasites from Senegal. We are now able to report that high-level resistance has evolved in three independent lines. Our ART resistance lines show all of the known in vitro phenotypic hallmarks of clinical ART resistance, but they are not K13 mutants! Remarkably, three independent selected lines each contain a different mutation in the gene PF3D7_1251200, which encodes Coronin, one of a family of WD-repeat proteins containing a beta propeller structure. The importance of Pf1251200 has already been demonstrated experimentally in our laboratory using CRISPR/Cas9 replacements. These findings demonstrate the existence of at least two distinct genetic mechanisms of ART resistance. Investigating the Pf1251200 mutants and their interactions with K13 and other genes will help elucidate the mechanism of action of artemisinin, which is still unknown, and perhaps more important will provide markers for early detection of non-K13 ART resistance in clinical settings in Africa as well as SE Asia where a significant proportion of ART-resistant isolates have no mutations in K13.

疟疾控制的最新进展减少了事件，并挽救了数百人的生命 成千上万的儿童，但有效的策略取决于包括特殊的措施的组合 强调基于青蒿素的组合疗法（ACTS）。预测的可怕后果 阿丁凝集素（艺术）抵抗的进化和传播在东南亚逐渐出现 艺术阻力迅速发展并迅速传播。抗艺术性决定居住在螺旋桨中 PFKELCH13基因座（K13）的域，这被认为可以促进蛋白质质量控​​制并调节应力 回答。重要的是，确定K13首先是通过五年的体外选择识别的 坦桑尼亚的抗性菌株测序； K13后来才在东南亚的田野中得到证实。 如果有抵抗艺术在非洲进行和蔓延的，那将是真正的灾难性。为什么没有开放 出于猜测，一种可能性是因为K13效应强烈取决于遗传背景， 非洲寄生虫和亚洲谱系之间存在显着的遗传差异。 但是，正如每位进化生物学家一样 知道 - 在非洲的抵抗决定者的演变中几乎不可避免地会产生。基于假设 四年前（K13报告之前），这种抵抗力可能取决于遗传背景 从塞内加尔选择独立的寄生虫线。我们现在能够报告高级 电阻在三个独立线上演变。我们的艺术阻力线显示了所有已知的 临床抗性的体外表型标志，但不是K13突变体！ 值得注意的是，三个独立的线条中都包含不同的基因突变 PF3D7_1251200，编码Coronin，Coronin是一个含有β螺旋桨的WD-重复蛋白的家族之一 结构。 PF1251200的重要性已经在我们的实验室中使用 CRISPR/CAS9替换。 这些发现证明了至少存在两种​​不同的艺术遗传机制 反抗。研究PF1251200突变体及其与K13和其他基因的相互作用将有助于 阐明阿秒蛋白的作用机理，但仍未知，也许更重要的 提供标志物，以早期检测非洲临床环境和SE Asia的非K13 ART耐药性 大量抗艺术的分离株在K13中没有突变。

项目成果

Natural selection constrains neutral diversity across a wide range of species.

• DOI: 10.1371/journal.pbio.1002112
• 发表时间: 2015-04
• 期刊: PLoS biology
• 影响因子: 9.8
• 作者: Corbett-Detig RB;Hartl DL;Sackton TB
• 通讯作者: Sackton TB

Genetic evidence that the Makira region in northeastern Madagascar is a hotspot of malaria transmission.

• DOI: 10.1186/s12936-016-1644-4
• 发表时间: 2016-12-20
• 期刊: Malaria journal
• 影响因子: 3
• 作者: Rice BL;Golden CD;Anjaranirina EJ;Botelho CM;Volkman SK;Hartl DL
• 通讯作者: Hartl DL

Genetic surveillance detects both clonal and epidemic transmission of malaria following enhanced intervention in Senegal.

• DOI: 10.1371/journal.pone.0060780
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Daniels R;Chang HH;Séne PD;Park DC;Neafsey DE;Schaffner SF;Hamilton EJ;Lukens AK;Van Tyne D;Mboup S;Sabeti PC;Ndiaye D;Wirth DF;Hartl DL;Volkman SK
• 通讯作者: Volkman SK

A broad analysis of resistance development in the malaria parasite.

对疟原虫耐药性发展的广泛分析。

• DOI: 10.1038/ncomms11901
• 发表时间: 2016-06-15
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Corey VC;Lukens AK;Istvan ES;Lee MCS;Franco V;Magistrado P;Coburn-Flynn O;Sakata-Kato T;Fuchs O;Gnädig NF;Goldgof G;Linares M;Gomez-Lorenzo MG;De Cózar C;Lafuente-Monasterio MJ;Prats S;Meister S;Tanaseichuk O;Wree M;Zhou Y;Willis PA;Gamo FJ;Goldberg DE;Fidock DA;Wirth DF;Winzeler EA
• 通讯作者: Winzeler EA

Genetic surveillance for monitoring the impact of drug use on Plasmodium falciparum populations.

• DOI: 10.1016/j.ijpddr.2021.07.004
• 发表时间: 2021-12
• 期刊: International journal for parasitology. Drugs and drug resistance
• 作者: Ndiaye YD;Hartl DL;McGregor D;Badiane A;Fall FB;Daniels RF;Wirth DF;Ndiaye D;Volkman SK
• 通讯作者: Volkman SK