Midgut Transcriptome and Proteome Analyses: Non-model Anopheline Malaria Vectors

中肠转录组和蛋白质组分析：非模型按蚊疟疾载体

基本信息

批准号：
8700629
负责人：
Rhoel David Ramos Dinglasan
金额：
$ 21.38万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8700629
关键词：
Address Adult Africa South of the Sahara African Anopheles Genus Anopheles gambiae Apical Attention Base Sequence Bioinformatics Biological Assay Biological Models Biology Blood Brush Border Cessation of life Clinical Communities Complement Complex Culicidae Data Data Analyses Development Disease Evolution Falciparum Malaria Gene Expression Profile Genes Genome Geographic Distribution Human Biology Hybrids Immune Infection Informatics Intervention Knowledge Laboratories Link Liquid Chromatography Malaria Mass Spectrum Analysis Mediating Membrane Midgut Mining Modeling Molecular Molecular Profiling Morbidity - disease rate Olives - dietary Organism Parasites Plasmodium Plasmodium falciparum Plasmodium vivax Preparation Proteins Proteome Proteomics Public Health Publishing Relative (related person)Research Sampling Scientist Side Solomon Islands System Technology Tissues Transcend Transcript Vaccines Vesicle Vivax Malaria burden of illness cellular microvillus combat comparative disorder risk feeding functional genomics genome annotation genome sequencing immune function innovation member mortality next generation novel protein expression public health relevance response sugar tandem mass spectrometry transcriptome sequencing transcriptomics transmission process vector vector mosquito

项目摘要

DESCRIPTION (provided by applicant): Development of Plasmodium parasites in the Anopheles mosquito is required for successful malaria parasite transmission. Although the greatest burden of disease occurs in Sub-Saharan Africa (SSA), malaria morbidity and mortality extends far beyond the African continent. Across the geographical distribution of known anopheline vectors of Plasmodium, morbidity and mortality are caused by both Plasmodium falciparum and Plasmodium vivax. In fact, P. vivax has the widest geographic distribution with 2.5 billion people at risk of the disease, and between 80 - 300 million clinical cases every year, including severe disease and death. Despite this tremendous public health burden, P. vivax research has received far less attention and support than efforts centered on P. falciparum. There is a recent heightened re-emphasis on studies aimed at elucidating the transmission biology of P. vivax. Unfortunately, little molecular and genome-scale information exists for vectors of P. vivax, which in some endemic regions outside of SSA are also vectors of P. falciparum. Moreover, although the molecular functions of the genome of the "model" African P. falciparum vector, Anopheles gambiae is relatively well studied, the substantial evolutionary divergence observed within anophelines limits its utility as the reference for the entire lineage, especially with respect to several P. vivax vectors. One of the major vectors of P. vivax and P. falciparum, and a potentially great emerging model system to understand the transmission biology of human Plasmodia through mosquitoes, is Anopheles farauti 1 (FAR1/AF1) and related species complex members in the Western Pacific. To better understand malaria transmission biology, it is imperative to more thoroughly examine the interaction between the parasite and mosquito tissues, particularly the mosquito midgut. In this project our overarching question is "What are the molecular determinants mediating Plasmodium ookinete-Anopheles midgut interactions?" Thus, we will perform comparative transcriptomic and proteomic analyses of mosquito midgut lumen expressed immune- related genes from the model (gambiae), the recently sequenced "emerging model" anopheline vector (FAR1) colony and wild-type farauti (AF1) mosquitoes, as well as the non-model but related species (An. punctulatus, AP). Our focus will be on the subset of immune-related genes that are differentially expressed as both transcript and protein in response to blood feeding and Plasmodium invasion. We will first focus on FAR1 infections with laboratory P. falciparum and complement these studies with subsequent P. vivax membrane feeding assays with FAR1/AF1/AP. It should be noted that the transcriptome assembly and proteomics analysis of the FAR1/AF1 mosquito midgut described in this project will not only strongly complement and enhance the vector-biology community's recent FAR1 sequencing effort and genome annotation for this species but will also greatly contribute to bringing our knowledge on vivax-anopheles interactions on par with that of falciparum-gambiae, thus allowing the two study systems to "reciprocally illuminate" one another.

描述（由申请人提供）：疟疾寄生虫的成功传播需要在按蚊中发育出疟原虫寄生虫。尽管最大的疾病负担发生在撒哈拉以南非洲（SSA），但疟疾发病率和死亡率远远超出了非洲大陆。在已知疟原虫按蚊载体的地理分布中，发病率和死亡率都是由恶性疟原虫和间日疟原虫引起的。事实上，间日疟原虫的地理分布最为广泛，有 25 亿人面临这种疾病的风险，每年有 80 - 3 亿临床病例，包括严重疾病和死亡。尽管存在巨大的公共卫生负担，但间日疟原虫研究所受到的关注和支持远少于针对恶性疟原虫的研究。最近重新强调了旨在阐明间日疟原虫传播生物学的研究。不幸的是，关于间日疟原虫载体的分子和基因组规模信息很少，在SSA以外的一些流行地区，间日疟原虫也是恶性疟原虫的载体。此外，虽然“模型”非洲恶性疟原虫载体冈比亚按蚊基因组的分子功能得到了相对充分的研究，但按蚊内部观察到的实质性进化差异限制了其作为整个谱系参考的效用，特别是在几个谱系方面。间日疟原虫载体。西太平洋按蚊 1 (FAR1/AF1) 和相关物种复合体成员是间日疟原虫和恶性疟原虫的主要媒介之一，也是了解人类疟原虫通过蚊子传播生物学的潜在重要新兴模型系统。为了更好地了解疟疾传播生物学，必须更彻底地检查寄生虫和蚊子组织（特别是蚊子中肠）之间的相互作用。在这个项目中，我们的首要问题是“介导动合疟原虫-按蚊中肠相互作用的分子决定因素是什么？”因此，我们将对来自模型（冈比亚）、最近测序的“新兴模型”按蚊载体（FAR1）群体和野生型法劳提（AF1）蚊子的蚊子中肠腔表达的免疫相关基因进行比较转录组和蛋白质组分析，如以及非模式但相关的物种（An. punctuatus，AP）。我们的重点将放在免疫相关基因的子集上，这些基因在响应血液喂养和疟原虫入侵时以转录物和蛋白质的形式差异表达。我们将首先关注实验室恶性疟原虫的 FAR1 感染，并通过随后的 FAR1/AF1/AP 间日疟原虫膜喂养测定来补充这些研究。值得注意的是，本项目中描述的 FAR1/AF1 蚊子中肠的转录组组装和蛋白质组学分析不仅将有力地补充和增强载体生物学界最近对该物种的 FAR1 测序工作和基因组注释，而且还将极大地促进将我们关于间日疟原虫相互作用的知识与关于恶性疟原虫-冈比亚疟原虫相互作用的知识结合起来，从而使两个研究系统能够“相互阐明”彼此。