Whole genome sequencing of Malagasy Duffy-independent P. vivax

马达加斯加非达菲依赖性间日疟原虫的全基因组测序

• 批准号: 8230462
• 负责人: PETER A. ZIMMERMAN
• 金额: $ 15.7万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-15 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8230462
• 关键词: Address; Admixture; African; Age; Alleles; Antigens; Area; Binding Proteins; Blood; Blood specimen; Blood typing procedure; Candidate Disease Gene; Characteristics; Child; Clinical; Culicidae; DNA; Data; Development; Disease; Epidemiology; Erythrocytes; Frequencies; Genes; Genetic Polymorphism; Genetic Variation; Genome; Genotype; Health; Human; Individual; Infection; Invaded; Laboratories; Life; Ligands; Linkage Disequilibrium; Madagascar; Malaria; Measures; Meiotic Recombination; Microscopy; Molecular Evolution; Mono-S; Parasites; Participant; Phenotype; Plasmodium vivax; Population; Population Sizes; Prevalence; Proteins; Recording of previous events; Relaxation; Research; Resistance; Reticulocytes; Risk; School-Age Population; Sequence Analysis; Shotgun Sequencing; Staging; Surface; Surveys; Technology; Testing; Time; Vivax Malaria; base; blood group; drug efficacy; experience; genome sequencing; in vivo; parasite invasion; pressure; public health relevance; response; trait; transmission process

DESCRIPTION (provided by applicant): Today 2.6 billion people live in areas of risk for Plasmodium vivax transmission; 106 - 313 million cases of vivax malaria annually. While Duffy blood group negativity has been considered to confer resistance to vivax malaria, our recent findings indicate that P. vivax has gained the capacity to infect erythrocytes and cause disease in Duffy-negative (Duffy(-)) people in Madagascar and suggests that P. vivax has acquire new erythrocyte infection mechanisms. In surveys of school-aged children 8.8% of asymptomatic Duffy(-) children (42/476) were P. vivax PCR-positive. Additionally, during surveys to assess in vivo efficacy of drugs recommended by the Madagascar Ministry of Health to treat malarial illness, we identified nine Duffy(-) people who had mono-infection P. vivax malaria (4.9% of 183 participants). We validated this unusual finding by microscopy to provide the first evidence for blood- stage development of P. vivax, including sexual-stage gametocytes necessary to continue the parasite lifecycle through mosquito transmission. Finally, flow cytometric analysis of erythrocytes from Malagasy study participants (n=43; many of whom experienced clinical P. vivax malaria) showed that Duffy(-) genotype and phenotype were 100% concordant, to indicate that the Duffy antigen was not cryptically expressed on the erythrocyte surface in genotypically Duffy(-) individuals. To begin understanding how P. vivax has gained capacity to infect Duffy(-) erythrocytes we propose to analyze the genome sequence of several P. vivax isolates extracted from blood samples of Duffy(+) and Duffy(-) Malagasy individuals, and the resulting data will allow us to address the following Specific Aims. Aim 1: Determine whether the same strains of P. vivax are responsible for infections of Duffy(-) and Duffy(+) individuals in Madagascar and reconstruct the population history of Malagasy P. vivax. Aim 2: Identify genes under recent positive selection in Malagasy P. vivax genomes. Our study of genetic diversity among multiple Malagasy P. vivax isolates by whole genome sequencing will allow us to determine how, or if the parasite population has partitioned in response to the important barrier imposed by Duffy negativity. By identifying loci under recent positive selection, our studies have strong potential to identify new parasite invasion ligands contributing to Duffy-independent erythrocyte invasion. PUBLIC HEALTH RELEVANCE:- Today 2.6 billion people live in areas of risk for Plasmodium vivax (Pv) transmission; 106 - 313 million cases of vivax malaria annually. Duffy blood group negativity has been considered to confer resistance to vivax malaria, however, our recent findings indicate that Pv has gained the capacity to infect erythrocytes and cause disease in Duffy-negative people in Madagascar, suggests that Pv has acquire new erythrocyte infection mechanisms. Through whole genome sequence analysis we will reveal differences between Pv strains infecting Duffy-negative vs. Duffy-positive people that will help identify genes evolving to expand the erythrocyte invasion capacity of Pv.

描述(申请人提供)：今天，26亿人生活在间日疟原虫传播的危险地区；每年有1.06亿-3.13亿间日疟病例。虽然Duffy血型阴性被认为是对间日疟的抵抗力，但我们最近的发现表明，间日疟原虫已经获得了感染红细胞并在马达加斯加Duffy阴性(Duffy(-))人群中致病的能力，并表明间日疟原虫获得了新的红细胞感染机制。在对学龄儿童的调查中，8.8%的无症状Duffy(-)儿童(42/476)间日疟原虫聚合酶链式反应阳性。此外，在评估马达加斯加卫生部推荐的治疗疟疾药物的体内疗效的调查中，我们确定了9名患有单感染间日疟的达菲(-)患者(183名参与者中有4.9%)。我们通过显微镜验证了这一不同寻常的发现，为间日疟原虫的血液期发育提供了第一个证据，包括通过蚊子传播延续寄生虫生命周期所必需的有性阶段配子体。最后，对马达加斯加研究参与者(n=43，其中许多人有间日疟临床经历)的红细胞进行的流式细胞仪分析显示，Duffy(-)型与表型100%一致，这表明Duffy(-)型个体的红细胞表面不存在Duffy抗原的隐蔽表达。为了开始了解间日疟原虫是如何获得感染达菲(-)红细胞的能力的，我们建议分析从达菲(+)和达菲(-)马达加斯加人的血液样本中提取的几个间日疟原虫分离株的基因组序列，所产生的数据将使我们能够解决以下具体目标。目的1：确定马达加斯加间日疟原虫感染达菲(-)和达菲(+)个体是否与同一株间日疟原虫有关，并重建间日疟原虫的种群历史。目的2：鉴定马达加斯加间日疟原虫基因组在最近正选择下的基因。我们通过全基因组测序对多个马达加斯加间日疟原虫分离株的遗传多样性进行研究，将使我们能够确定寄生虫种群是如何分裂的，或者是否对达菲负性所造成的重要障碍做出了反应。通过识别最近正选择下的基因座，我们的研究具有很强的潜力来识别新的寄生虫入侵配体，这些配体有助于Duffy非依赖的红细胞入侵。 与公共卫生相关：-今天有26亿人生活在间日疟原虫传播的风险地区；每年有1.06亿-3.13亿间日疟病例。Duffy血型阴性一直被认为是对间日疟的抵抗力，然而，我们最近的发现表明，在马达加斯加Duffy阴性的人中，PV已经获得了感染红细胞并导致疾病的能力，这表明PV获得了新的红细胞感染机制。通过全基因组序列分析，我们将揭示感染Duffy阴性和Duffy阳性人群的PV毒株之间的差异，这将有助于识别进化为扩大PV红细胞侵袭能力的基因。

项目成果

Comparative analysis of field-isolate and monkey-adapted Plasmodium vivax genomes.

• DOI: 10.1371/journal.pntd.0003566
• 发表时间: 2015-03
• 期刊: PLoS neglected tropical diseases
• 影响因子: 3.8
• 作者: Chan ER;Barnwell JW;Zimmerman PA;Serre D
• 通讯作者: Serre D

De novo assembly of a field isolate genome reveals novel Plasmodium vivax erythrocyte invasion genes.

• DOI: 10.1371/journal.pntd.0002569
• 发表时间: 2013
• 期刊: PLoS neglected tropical diseases
• 影响因子: 3.8
• 作者: Hester J;Chan ER;Menard D;Mercereau-Puijalon O;Barnwell J;Zimmerman PA;Serre D
• 通讯作者: Serre D