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.