An.gambiae immune signaling gene SNPs and natural P. falciparum infection

冈比亚按蚊免疫信号基因SNPs与天然恶性疟原虫感染

基本信息

批准号：
8197064
负责人：
GREGORY C. LANZARO
金额：
$ 65.44万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-15 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8197064
关键词：
Address Affect Africa South of the Sahara Alleles Anopheles gambiae Cameroon Cell Line Cells Central Africa Chemicals Chromosome inversion Code Codon Nucleotides Country Culicidae Data Development Exhibits Gene Conversion Gene Targeting Gene Transfer Techniques Genes Genetic Genetic Polymorphism Genetic Variation Genotype Geographic Distribution Goals Human Immune Immune response Immune system Immunity In Vitro Infection Insulin Karyotype Laboratory Study Light Malaria Mali Maps Mediating Methods Midgut Molecular Nature O&apos nyong-nyong virus Parasites Phenotype Plasmodium falciparum Population Population Genetics Predisposition Protocols documentation Regulation Sampling Screening procedure Signal Pathway Signal Transduction Single Nucleotide Polymorphism Site Techniques Testing Transfection Translations Work base forest gene function genetic analysis in vivo inhibitor/antagonist insight interest knock-down novel overexpression protein function protein structure public health relevance reproductive response transmission process vector

项目摘要

DESCRIPTION (provided by applicant): Significant data implicate the Toll/imd, insulin, and TGF-¿ signaling pathways in the regulation of malaria parasite development in the Anopheles gambiae midgut. However, no data are available to confirm that these signaling pathways regulate parasite development in nature. We have chosen to address this issue using a SNP association mapping approach with field-collected mosquitoes from Mali and Cameroon, countries with high malaria transmission that include the geographic distribution of the most genetically diverged An. Gambiae populations yet described. We will pursue this goal through two specific aims. The first Aim includes population genetic analyses. Plasmodium falciparum infected and uninfected An. gambiae will be collected from sites in Mali and Cameroon. These sites were carefully selected to include all of the genetic diversity known to exist in the region (three molecular forms and chromosome inversion polymorphism). Mosquito samples will be grouped by site with respect to infection, molecular form and karyotype. Each mosquito will then be genotyped for roughly 384 immune signaling gene SNPs and those SNPs correlated with infection identified by association mapping. This analysis will identify SNPs putatively involved in susceptibility to P. falciparum infection. In the second Aim, selected SNPs of interest will be analyzed to determine their effects on mosquito protein function and on susceptibility to P. falciparum infection. Specifically, we will utilize immortalized An. gambiae cell lines and a combination of over expression, knock-down, and gene conversion techniques to confirm predictions of function from sequence and to prioritize SNPs for in vivo studies. For the in vivo studies, we will use two genetically distinct strains of An. gambiae, which have been genotyped for the SNPs of interest. We will use inhibitors and transfection (knockdown, O'nyong-nyong infectious clone mediated overexpression) protocols to mimic the effects of SNP-containing alleles on P. falciparum development in infected mosquitoes. Our work will take functional data from our lab and from the labs of our colleagues to examine the importance of the selected immune signaling pathways in field-collected mosquitoes. In this light, the studies proposed herein will facilitate selection of appropriate gene targets for transgenesis strategies and provide critical new insights into the population genetics of immunity in An. gambiae that to our knowledge are not currently available. PUBLIC HEALTH RELEVANCE: The mosquito Anopheles gambiae transmits the human malaria parasite Plasmodium falciparum in sub-Saharan Africa. Many laboratory studies have focused on how the mosquito immune system responds to and destroys these parasites, but there is little to no information on whether these responses are important in nature. Our studies will identify responses that are important in natural populations of An. gambiae with the long-term goal that this information can contribute to novel malaria control methods.

描述（由申请人提供）：重要数据意味着 Toll/imd、胰岛素和 TGF-¿冈比亚按蚊中肠疟疾寄生虫发育调节中的信号通路然而，没有数据可以证实这些信号通路在自然界中调节寄生虫发育，我们选择使用现场收集的 SNP 关联作图方法来解决这个问题。来自马里和喀麦隆这两个疟疾传播率高的国家的蚊子，其中包括迄今描述的遗传差异最大的冈比亚蚊子种群。我们将通过两个具体目标来实现这一目标。将从马里和喀麦隆的地点收集恶性疟原虫感染和未感染的按蚊，这些地点包括该地区已知存在的所有遗传多样性（三种分子形式和染色体倒位多态性）。然后根据感染部位、分子形式和核型对每只蚊子进行大约 384 个免疫信号基因 SNP 的基因分型，以及通过关联识别与感染相关的 SNP。该分析将识别可能与恶性疟原虫感染易感性有关的 SNP。利用永生化冈比亚按蚊细胞系和过度表达、敲低和基因转换技术的组合来确认序列的功能预测并确定体内研究的 SNP 优先级。在体内研究中，我们将使用两种遗传上不同的冈比亚按蚊菌株，它们已针对感兴趣的 SNP 进行了基因分型。我们将使用抑制剂和转染（敲低、O'nyong-nyong 感染性克隆介导的过度表达）方案来模拟受感染的蚊子中恶性疟原虫发育中含有 SNP 的等位基因我们的工作将从我们的实验室和同事的实验室获取功能数据，以检查所选免疫信号通路的重要性。有鉴于此，本文提出的研究将有助于为转基因策略选择适当的基因靶标，并为冈比亚按蚊的免疫群体遗传学提供重要的新见解，而据我们所知，目前尚无法获得这些见解。公共卫生相关性：冈比亚按蚊在撒哈拉以南非洲传播人类疟原虫恶性疟原虫，许多实验室研究都集中在蚊子免疫系统如何响应并消灭这些寄生虫，但几乎没有关于这些反应是否有效的信息。我们的研究将确定对冈比亚自然种群重要的反应，长期目标是这些信息有助于新的疟疾控制方法。