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: 9751186
• 负责人: Michael T Ferdig
• 金额: $ 236.65万
• 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 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751186
• 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; base; blood treatment; crosslink; data archive; 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.

抽象的 遗传杂交以及连锁映射的遗传杂交为您提供了出色的成功方法 定位生物医学重要特征的遗传决定因素，例如耐药性和宿主特异性 恶性疟疾疟疾。最初使用黑猩猩进行疟原虫交叉，但使用 这些灵长类动物现在已经不可能了。该计划项目赠款（P01）的总体目标是利用 尖端技术使我们能够上台恶性疟原虫实验性遗传杂交和 分离大量独特的重组后代。我们使用注入的人肝小鼠这样做 与人类红细胞（FRG HUHEP/HURBC小鼠）。我们将使用这项技术来解决 抗凝集素耐药蛋白（ART-R）的出现和传播构成的新兴健康威胁以及更多 最近的抗酮抵抗寄生虫。 最近有很多关于识别kelch13基因中的编码突变的识别 与ART-R密切相关。但是，对该基因的功能以及如何了解 kelch13中的突变产生了广泛的电阻水平和健身效应，以及这些影响如何 由基因组的其他结构或调节变化弥补。此外，还有艺术的证据 kelch13中没有突变的R，并且特定的遗传背景对于ART-R允许。我们将使用 有针对性的实验遗传交叉（i）剖析ART-R的遗传复杂性，（ii）阐明 kelch13，（iii）定义控制ART-R的调节剂和伴侣基因，（iv）确定遗传基础 新兴的载管抗性。 该项目位于三个地点（巴黎圣母院，西雅图和圣安东尼奥），每个位置都有一项研究 由核心设施支持的项目，在巴黎圣母院拥有行政核心。三个研究核心 支持三个个人研究项目中每个项目的任务，并相互依靠 后代线，测序和互补数据分析。每个研究项目都有自己的独立 仅研究问题，但是项目之间的信息和试剂的流动显着增强了 完全利用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门德利的基本方面的理解 恶性疟原虫中的遗传学，使我们能够进一步优化产生重组后代的方法。这 重组后代将以RP02的耐药性和竞争性生长表型为特征 以及RP03的转录本，蛋白质和代谢物丰度的变化，并在CoreC的支持下。 表型，序列和系统遗传数据将由Core B集成，Core B将进行分析 并确保所有三个研究项目都可以访问存档数据。