Experimental Plasmodium falciparum crosses using human-liver chimeric mice

使用人肝嵌合小鼠进行实验性恶性疟原虫杂交

• 批准号: 8959920
• 负责人: Ashley M Vaughan
• 金额: $ 21.2万
• 依托单位: SEATTLE BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-11-01 至 2016-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8959920
• 关键词: Anopheles Genus; Anti-malarial drug resistance; Antimalarials; Artemisinins; Biomedical Research; Blood; Chloroquine; Chloroquine resistance; Chromosome Mapping; Chromosomes, Human, Pair 7; Combined Modality Therapy; Communities; Culicidae; Data; Development; Disease; Drug resistance; Erythrocytes; Exhibits; Experimental Genetics; Female; Frequencies; Future; Generations; Genes; Genetic; Genetic Crosses; Genetic Determinism; Genetic Population Study; Genetic Recombination; Genetic Research; Genetic study; Genotype; Growth; Health; Hepatocyte; Human; Infection; Ingestion; Insecticide Resistance; Intervention; Liver; Maintenance; Malaria; Maps; Methodology; Modality; Modeling; Morbidity - disease rate; Mus; Pan Genus; Parasites; Partner in relationship; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Plasmodium; Plasmodium falciparum; Point Mutation; Predisposition; Process; Production; Publishing; Quantitative Trait Loci; Recombinants; Recovery; Reproduction; Resistance; Salivary Glands; Sporozoites; Staging; System; Technology; Testing; Transgenic Organisms; United States National Institutes of Health; Validation; Variant; artemisinine; asexual; base; genetic analysis; genetic manipulation; improved; killings; mortality; mouse model; novel; pathogen; programs; resistant strain; response; tool; trait; vector mosquito

 DESCRIPTION (provided by applicant): Plasmodium parasites cause the disease malaria, which kills close to one million people per year. The disease is formidable, due to the spread of drug-resistant parasite strains, insecticide-resistant mosquito vectors and the inherent difficultis in the implementation and maintenance of effective control programs. Today, emerging resistance to artemisinin combination therapy, which is the standard drug treatment for uncomplicated malaria, poses a significant threat to malaria control. Understanding the genetic basis of antimalarial drug resistance is key and will inform development of new interventions. Experimental crosses between malaria parasite strains are the most powerful way to determine the genetic determinants of phenotypes, and importantly, have been successfully used to determine the genetic basis of P. falciparum drug resistance in the past. To date however, only three P. falciparum experimental crosses have been carried out due to the enormous practical and financial hurdles associated with the process, which cannot be performed in humans, and thus used splenectomized chimpanzees. The genetic analysis of recombinant progeny from a well-conceived cross can be extremely powerful for pinpointing the genetic loci determining a phenotypic trait and has important advantages over population genetic studies. Given the recent decision by the NIH to cease the use of chimpanzees in biomedical research, experimental crosses will no longer be available, effectively rendering future forward genetic studies impossible. However, we have recently shown that a mouse harboring human hepatocytes (the FRG KO huHep mouse) infused with human red blood cells can support P. falciparum sporozoite infection, complete liver stage development as well as the transition to asexual blood stage replication. We hypothesize that the FRG KO huHep mouse can be utilized successfully for routine and robust P. falciparum experimental crosses, thereby replacing the previously essential chimpanzee host. We have data to show that we are able to perform experimental genetic crosses utilizing this model and the crosses generated independent recombinant progeny that we will use for analysis of recombination as well as genotypes and associated phenotypes. Furthermore, we have also carried out an experimental cross with a recent field isolate resistant to artemisinin, allowing us to study the basis of artemisinin resistance. Combined with rapidly improving technologies, such as parasite genetic manipulation, high throughput phenotyping and Systems Genetics, a powerful model for malaria genetic research is now within reach.

 描述（由适用提供）：疟原虫引起疾病疟疾，每年杀死近100万人。由于抗药性寄生虫菌株，耐杀虫剂的蚊子媒介以及继承在实施和维持有效控制程序方面的困难，该疾病是可以形式化的。如今，新兴的抗毒素联合疗法的抗性是对疟疾的标准药物治疗，对疟疾的控制构成了重大威胁。了解抗性药物耐药性的遗传基础是关键，并将为新干预措施的发展提供信息。疟疾寄生虫菌株之间的实验杂交是确定表型遗传决定剂的最有力方法，重要的是，已经成功地用于确定过去的恶性疟原虫耐药性的遗传基础。然而，迄今为止，由于与该过程相关的巨大实践障碍和财务障碍，因此仅进行了三个恶性疟原虫实验十字架，而这些障碍无法在人类中进行，因此使用了脾脏切除的黑猩猩。重组进步的遗传分析来自良好的十字架，对于确定表型性状的遗传基因座的遗传性可能非常有力，并且在人群遗传研究中具有重要的优势。鉴于NIH最近决定停止使用黑猩猩在生物医学研究中的使用，因此将不再可用实验十字架，从而有效地使未来的前瞻性遗传研究不可能。但是，我们最近表明，感染了人类红细胞的人类肝细胞（FRG KO Huhep小鼠）的小鼠可以支持恶性疟原虫孢子岩感染，完整的肝脏阶段发育以及向无性血液阶段复制的过渡。我们假设可以将FRG KO Huhep小鼠成功用于常规和强大的恶性疟原虫实验十字架，从而取代先前必需的黑猩猩宿主。我们有数据表明，我们能够使用该模型执行实验性遗传杂交，并且杂交产生了独立的重组后代，我们将用于分析重组以及基因型和相关表型。此外，我们还与最近对青蒿素抗性具有抗性的田间分离株进行了实验十字，使我们能够研究青蒿素耐药性的基础。结合迅速改进的技术，例如寄生虫遗传操纵，高通量表型和系统遗传学，现在可以实现疟疾遗传研究的强大模型。