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（由申请人提供）：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 - 3.13亿例间日疟原虫疟疾病例。虽然Duffy血型阴性被认为赋予了对间日疟的抗性，但我们最近的研究结果表明，间日疟已经获得了感染红细胞并在马达加斯加Duffy阴性（Duffy（-））人群中引起疾病的能力，并表明间日疟已经获得了新的红细胞感染机制。在学龄儿童的调查中，8.8%的无症状Duffy（-）儿童（42/476）为间日疟原虫PCR阳性。此外，在评估马达加斯加卫生部推荐的治疗疟疾疾病的药物的体内疗效的调查中，我们确定了9名患有单一感染间日疟原虫疟疾的达菲（-）人（183名参与者的4.9%）。我们通过显微镜验证了这一不寻常的发现，为间日疟原虫的血液阶段发育提供了第一个证据，包括通过蚊子传播继续寄生虫生命周期所需的有性阶段配子细胞。最后，对来自马达加斯加研究参与者（n=43;其中许多人经历了临床间日疟原虫疟疾）的红细胞进行流式细胞术分析，结果显示Duffy（-）基因型和表型100%一致，表明Duffy抗原在基因型Duffy（-）个体的红细胞表面上没有隐蔽表达。为了开始了解间日疟原虫如何获得感染达菲（-）红细胞的能力，我们建议分析从达菲（+）和达菲（-）马达加斯加个体的血液样本中提取的几种间日疟原虫分离株的基因组序列，所得数据将使我们能够解决以下特定目标。目标1：确定马达加斯加Duffy（-）和Duffy（+）个体的感染是否与间日疟原虫相同，并重建马达加斯加间日疟原虫的种群历史。目的2：在马达加斯加间日疟原虫基因组中鉴定最近正选择的基因。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的研究将使我们能够确定寄生虫种群如何或是否已对达菲阴性所施加的重要障碍做出反应。通过确定最近的正选择下的位点，我们的研究有很大的潜力，以确定新的寄生虫入侵配体的达菲独立的红细胞入侵。 与公共卫生的关系：今天，有26亿人生活在间日疟原虫（Pv）传播的风险地区;每年有1.06亿至3.13亿例间日疟原虫疟疾病例。Duffy血型阴性一直被认为是间日疟的抵抗力，然而，我们最近的研究结果表明，Pv已经获得了感染红细胞的能力，并在马达加斯加Duffy阴性人群中引起疾病，这表明Pv已经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阴性与Duffy阳性人群的Pv菌株之间的差异，这将有助于识别进化以扩大Pv红细胞侵袭能力的基因。

项目成果

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

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