Developing genetic tools for a new model of infection and immunity

开发新感染和免疫模型的遗传工具

基本信息

批准号：
8849839
负责人：
Ioannis Eleftherianos
金额：
$ 23.46万
依托单位：
GEORGE WASHINGTON 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：
2014-05-16 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8849839
关键词：
Adolescent Agriculture Animal Model Antigen Targeting Area Bacteria Biological Models Biology Biomedical Research Cadaver Caenorhabditis elegans Candidate Disease Gene Cellular biology Child Chromosomes Communicable Diseases DNA Development Double-Stranded RNA Drosophila genus Drosophila melanogaster Drug Targeting Drug resistance Elderly Elements Event Exploratory/Developmental Grant Food Foundations Future Gene Expression Gene Transfer Techniques Generations Genes Genetic Germ Lines Health Host Defense Mechanism Human Immune response Immunity Individual Infection Insecta Intestines Invaded Investigation Life Life Cycle Stages Microbe Microinjections Modeling Molecular Molecular Genetics Morbidity - disease rate Natural Immunity Nature Nematoda Nematode infections Organism Parasites Parasitic nematode Pathogenicity Pharmaceutical Preparations Plague Population Positioning Attribute Pregnant Women Process Production Protocols documentation RNA Interference Recovery Reporter Reporter Genes Research Retrotransposon Role School-Age Population Signal Transduction Source Staging System Techniques Technology Time Transfection Transgenes Vaccine Antigen Vaccines Virulence Factors World Bank Xenorhabdus luminescens base disability-adjusted life years exhaust gain of function gene function immune function improved killings knock-down life history method development novel pathogen promoter response reverse genetics tool vector

项目摘要

DESCRIPTION (provided by applicant): Parasitic nematode (PN) infections remain a major threat to human health worldwide, with more than 1 billion people infected. Children, pregnant women, and the elderly are particularly susceptible to morbidity from nematode infection. Control strategies are restricted to periodic de-worming of infected individuals, which is limited by rapid re-infection rates and the development of drug resistant worm populations. There are no vaccines available for PN infections in humans. Development of new drugs and vaccines will require a better understanding of PN biology, particularly the infective process and the host's immune response to infection. The requirement of an obligate host and the lack of good animal models have limited investigations into mechanisms by which PNs infect and usurp the host immune response. While the free living nematode Caenorhabditis elegans is an excellent model for nematode development, it is not a parasite. The insect parasitic nematode Heterorhabditis bacteriophora (Hb) offers potential as a tractable model of PN infection that has many of the advantages of C. elegans, including easy culture and manipulation of all life history stages. However, many of the powerful genetic tools, like transgenesis and RNA interference, have not been fully developed in Hb, restricting its use as a widespread model. We propose to develop these genetic tools in Hb with a focus on targeting the infective juvenile and early parasitic stages in order to investigate infection mechanisms. We will take two parallel but independent approaches to develop these tools that will provide a foundation for future investigation into the mechanism of PN infection. In Aim 1, we propose to develop a transfection protocol for Hb via germ-line microinjection of a piggybac retrotransposon-based integrating vector to introduce the reporter transgenes into Hb chromosomes. In Aim 2, we will develop a robust RNAi protocol for 2nd generation Hb hermaphrodites. By focusing on 2nd generation hermaphrodites we are ideally positioned to probe the biology of PN infection, as the progeny of this stage are the infective stage. Development of methods for gene knock down (reverse genetics) and transfection in this novel model nematode would represent a significant advance for PN research, and will allow investigation of PN gene function during infection for the first time.

描述（由申请人提供）：寄生线虫（PN）感染仍然是对全球人类健康的主要威胁，感染了超过10亿人。儿童，孕妇和老年人特别容易受到线虫感染发病率的影响。控制策略仅限于感染个体的定期驱动，这受到快速再感染率和耐药蠕虫种群的发展的限制。人类没有可用于PN感染的疫苗。新药和疫苗的开发将需要更好地了解PN生物学，尤其是感染过程以及宿主对感染的免疫反应。专性宿主和缺乏良好动物模型的要求对PNS感染和篡夺宿主免疫反应的机制的研究有限。虽然秀丽隐杆线虫的自由线虫是线虫发展的绝佳模型，但它不是寄生虫。昆虫寄生虫线虫异源性炎菌（HB）提供了一种潜在的PN感染模型，该模型具有秀丽隐杆线虫的许多优势，包括简单的培养和对所有生活史阶段的操纵。但是，许多强大的遗传工具，例如转基因和RNA干扰，在HB中尚未完全开发，从而限制了其作为广泛模型的使用。我们建议在HB中开发这些遗传工具，重点是针对感染性少年和早期寄生阶段，以研究感染机制。我们将采用两种平行但独立的方法来开发这些工具，为将来研究PN感染机制提供基础。在AIM 1中，我们建议通过基于PiggyBac逆转录转座的种系微注射为HB制定转染方案，以将报告基因转基因引入HB染色体中。在AIM 2中，我们将为第二代HB Hermaphrodites开发强大的RNAi方案。通过关注第二代雌雄同体，我们可以理想地探究PN感染的生物学，因为该阶段的后代是感染阶段。在这种新型模型线虫中，基因敲低方法（反向遗传学）和转染的发展将代表PN研究的重大进步，并将首次研究在感染过程中PN基因功能。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Identification of candidate infection genes from the model entomopathogenic nematode Heterorhabditis bacteriophora.

DOI：
10.1186/s12864-016-3468-6
发表时间：
2017-01-03
期刊：
BMC genomics
影响因子：
4.4
作者：
Vadnal J;Ratnappan R;Keaney M;Kenney E;Eleftherianos I;O'Halloran D;Hawdon JM
通讯作者：
Hawdon JM

Parasitic Nematode Immunomodulatory Strategies: Recent Advances and Perspectives.

DOI：
10.3390/pathogens5030058
发表时间：
2016-09-14
期刊：
Pathogens (Basel, Switzerland)
影响因子：
0
作者：
Cooper D;Eleftherianos I
通讯作者：
Eleftherianos I

Evolution and Function of Thioester-Containing Proteins and the Complement System in the Innate Immune Response.

DOI：
10.3389/fimmu.2017.00759
发表时间：
2017
期刊：
Frontiers in immunology
影响因子：
7.3
作者：
Shokal U;Eleftherianos I
通讯作者：
Eleftherianos I

Refined ab initio gene predictions of Heterorhabditis bacteriophora using RNA-seq.

使用 RNA-seq 精炼异杆菌基因预测。

DOI：
10.1016/j.ijpara.2018.02.001
发表时间：
2018
期刊：
International journal for parasitology
影响因子：
4
作者：
Vadnal,Jonathan;Granger,OliviaG;Ratnappan,Ramesh;Eleftherianos,Ioannis;O'Halloran,DamienM;Hawdon,JohnM
通讯作者：
Hawdon,JohnM

Entomopathogenic and plant pathogenic nematodes as opposing forces in agriculture.