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（由应用程序提供）：重要数据暗示在调节冈比亚邦比亚的疟原虫发育调控中，TOLL/IMD，胰岛素和TGF-ood信号通路。但是，无法确认这些信号通路调节自然界的寄生虫发育。我们选择使用SNP协会映射方法与马里和喀麦隆，疟疾传播高的国家（包括遗传上最有分歧的地理分布）一起解决此问题。冈比亚人口尚未描述。我们将通过两个具体目标来追求这一目标。第一个目的包括种群遗传分析。恶性疟原虫感染和未感染的冈比亚将从马里和喀麦隆的地点收集。仔细选择了这些位点，以包括该区域中已知存在的所有遗传多样性（三种分子形式和染色体反转多态性）。蚊子样品将按照感染，分子形式和核型进行分组。然后，将对大约384个免疫信号基因SNP和与通过关联映射确定的感染相关的SNP进行基因分型。该分析将确定涉及恶性疟原虫感染的敏感性的SNP。在第二个目标中，将分析选定的感兴趣SNP，以确定其对蚊子蛋白功能的影响以及对恶性疟原虫感染的易感性。具体而言，我们将利用永生的AN。冈比亚细胞系以及过度表达，敲低和基因转化技术的结合，以确认从序列的功能预测并优先考虑SNP进行体内研究。对于体内研究，我们将使用两种遗传上不同的An菌株。冈比亚，已针对感兴趣的SNP进行了基因分型。我们将使用抑制剂和翻译（敲低，O'Nyong-Nyong感染性克隆介导的过表达）方案来模仿含SNP的等位基因对受感染蚊子的恶性疟原虫发育的影响。我们的工作将从我们的实验室和同事的实验室中获取功能数据，以检查所选免疫信号通路在现场收集的蚊子中的重要性。据此，本文提出的研究将促进选择适当的基因靶标，以进行转化策略，并为人群提供关键的新见解。免疫的遗传学。据我们所知，冈比亚目前尚不可用。公共卫生相关性：蚊子冈比亚蚊子传播了撒哈拉以南非洲的人类疟原虫恶性疟原虫。许多实验室研究集中于蚊子免疫系统如何应对和破坏这些寄生虫，但是几乎没有关于这些反应在本质上是否重要的信息。我们的研究将确定在AN自然种群中很重要的反应。冈比亚的长期目标是这些信息可以有助于新型疟疾控制方法。