Harnessing the power of experimental genetic crosses and systems genetics to probe drug resistance in malaria

利用实验遗传杂交和系统遗传学的力量来探测疟疾的耐药性

• 批准号: 10216641
• 负责人: Michael T Ferdig
• 金额: $ 200.45万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10216641
• 关键词: Address; Affect; Antimalarials; Artemisinins; Biochemical Pathway; Chromosome Mapping; Code; Collaborations; Combined Modality Therapy; Communities; Complex; Comprehension; Core Facility; Coupled; Data; Data Analyses; Drug Targeting; Drug resistance; Ensure; Erythrocytes; Evolution; Experimental Genetics; Falciparum Malaria; Gene Mutation; Generations; Genes; Genetic; Genetic Crosses; Genetic Determinism; Genetic Recombination; Genetic Transcription; Genome; Goals; Grant; Growth; Health; Human; Individual; Insecticides; Link; Liver; Location; Malaria; Measurement; Methods; Mosquito Control; Mus; Mutation; Pan Genus; Parasite resistance; Parasites; Partner in relationship; Pharmaceutical Preparations; Phenotype; Plasmodium; Plasmodium falciparum; Primates; Process; Program Research Project Grants; Proteome; Proteomics; Quantitative Trait Loci; Reagent; Recombinants; Research; Research Project Grants; Resistance; Role; Site; Specificity; Structure; System; Technology; Time; Transcript; Variant; archive data; archived data; base; blood treatment; crosslink; disorder control; effective therapy; fitness; gene function; gene interaction; genetic analysis; genetic linkage analysis; improved; innovative technologies; insight; metabolome; metabolomics; novel strategies; protein metabolite; resistant Plasmodium falciparum; response; synergism; technology development; trait; transcriptome; transcriptomics; vector control; virtual

ABSTRACT Genetic crosses coupled with linkage mapping have provided an outstandingly successful approach for locating the genetic determinants of biomedically important traits such as drug resistance and host specificity in P. falciparum malaria. Plasmodium crosses were originally conducted using chimpanzees, but crosses using these primates are now no longer possible. The overall goal of this Program Project grant (P01) is to leverage cutting edge technology that enables us to stage Plasmodium falciparum experimental genetic crosses and isolate large numbers of unique recombinant progeny. We do this using a human-liver chimeric mouse infused with human red blood cells (the FRG huHep/huRBC mouse). We will use this technology to address the emerging health threat posed by the emergence and spread of artemisinin resistant (ART-R) and more recently piperaquine resistant parasites. There has been much fanfare recently about the identification of coding mutations in the kelch13 gene that are strongly associated with ART-R. However, very little is known about the function of this gene and how mutations in kelch13 generate a wide range of resistance levels and fitness effects, and how these effects are compensated by other structural or regulatory changes in the genome. Furthermore, there is evidence of ART- R without mutations in kelch13, and that particular genetic backgrounds are permissive for ART-R. We will use targeted experimental genetic crosses to (i) dissect the genetic complexity of ART-R, (ii) clarify the role of kelch13, (iii) define the regulators and partner genes that control ART-R, and (iv) determine the genetic basis of emerging piperaquine resistance. The project is based in three locations (Notre Dame, Seattle and San Antonio), each with one Research Project supported by a Core facility, with an Administrative Core in Notre Dame. The three research Cores support the tasks of each of the three individual Research Projects, and rely on each other for the generation of progeny lines, sequencing and complementary data analysis. Each Research Project has its own stand- alone research questions, but the flow of information and reagents among projects significantly enhances the potential for discovery that fully leverages the P01 framework. Genetic crosses will be conducted by Core A, while RP01 will increase our understanding of the fundamental aspects of sexual recombination and Mendelian genetics in P. falciparum, allowing us to further optimize methods for generating recombinant progeny. The recombinant progeny will be characterized for drug resistance and competitive growth phenotypes by RP02 and for variation in transcript, protein and metabolite abundance by RP03, with support from Core C. The phenotype, sequence and systems genetic data will be integrated by Core B, which will both conduct analyses and ensure that the archived data will be accessible to all three Research Projects.

摘要 遗传杂交与连锁作图相结合提供了一种非常成功的方法 确定重要生物医学性状的遗传决定因素，如抗药性和宿主专一性 恶性疟原虫。疟原虫杂交最初是用黑猩猩进行的，但用 这些灵长类动物现在已经不可能了。本计划项目赠款的总体目标(P01)是利用 尖端技术，使我们能够上演恶性疟原虫实验性遗传杂交和 分离出大量独特的重组后代。我们使用一只人肝嵌合小鼠来完成这项工作 与人红细胞(FRG huHep/huRBC小鼠)进行比较。我们将使用这项技术来解决 青蒿素抗药性(ART-R)的出现和蔓延造成的新的健康威胁 最近，对哌喹产生抗药性的寄生虫。 最近关于kelch13基因编码突变的鉴定大张旗鼓，这些突变是 与ART-R密切相关。然而，人们对该基因的功能及其如何发挥作用知之甚少。 Kelch13的突变会产生广泛的抗性水平和适应效果，以及这些影响是如何产生的 通过基因组中的其他结构或调节变化来补偿。此外，还有艺术的证据- 在kelch13中没有突变，并且特定的遗传背景对ART-R是允许的。我们将使用 有针对性的实验遗传交叉以(I)剖析ART-R的遗传复杂性，(Ii)阐明 Kelch13，(Iii)定义控制ART-R的调节基因和伙伴基因，以及(Iv)确定遗传基础 新出现的哌喹抗药性。 该项目以三个地点(巴黎圣母院、西雅图和圣安东尼奥)为基础，每个地点都有一个研究 由核心设施支持的项目，在巴黎圣母院有一个管理核心。三大研究核心 支持三个独立研究项目的每一个任务，并相互依赖 子代品系、测序和互补数据分析。每个研究项目都有自己的立场- 单独的研究问题，但项目之间的信息和试剂流动显著增强了 充分利用P01框架的发现潜力。遗传杂交将由核心A进行， 虽然RP01将增加我们对性重组和孟德尔式的基本方面的理解 恶性疟原虫的遗传学，使我们能够进一步优化产生重组后代的方法。这个 重组子代的抗药性和竞争性生长表型将由RP02鉴定 以及在Core C的支持下，RP03的转录本、蛋白质和代谢物丰度的变化。 表型、序列和系统遗传数据将由Core B整合，两者都将进行分析 并确保所有三个研究项目都可以访问存档的数据。