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（申请人提供）：疟原虫寄生虫引起疟疾，每年导致近100万人死亡。这种疾病是可怕的，由于抗药性寄生虫株的传播，抗药性蚊子载体和固有的困难，在实施和维持有效的控制方案。今天，对青蒿素综合疗法-治疗无并发症疟疾的标准药物疗法-出现的抗药性对疟疾控制构成重大威胁。了解抗疟药物耐药性的遗传基础是关键，将为制定新的干预措施提供信息。疟疾寄生虫株之间的实验杂交是确定表型的遗传决定因素的最有效方法，重要的是，过去已成功用于确定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的实践和财政障碍，仅进行了三次恶性疟原虫实验性杂交，该过程无法在人类中进行，因此使用了脾切除的黑猩猩。从一个精心设计的杂交重组后代的遗传分析可以是非常强大的精确定位的遗传位点决定的表型性状，并具有重要的优势，人口遗传研究。鉴于美国国立卫生研究院最近决定停止在生物医学研究中使用黑猩猩，实验性杂交将不再可用，这实际上使未来的前瞻性遗传研究变得不可能。 然而，我们最近已经表明，输注人红细胞的携带人肝细胞的小鼠（FRG KO huHep小鼠）可以支持恶性疟原虫子孢子感染，完成肝脏阶段发育以及向无性血液阶段复制的过渡。我们假设FRG KO huHep小鼠可成功用于常规和稳健的恶性疟原虫实验性杂交，从而取代先前必需的黑猩猩宿主。我们有数据表明，我们能够利用该模型进行实验性遗传杂交，并且杂交产生了独立的重组后代，我们将使用这些后代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耐药的田间分离株进行了实验性杂交，使我们能够研究青蒿素耐药性的基础。再加上寄生虫基因操作、高通量表型分析和系统遗传学等迅速改进的技术，一个强大的疟疾基因研究模型现在已经触手可及。