Genome wide assessment of essential genes in Trypanosoma brucei

布氏锥虫必需基因的全基因组评估

• 批准号: 8204844
• 负责人: KENNETH D STUART
• 金额: $ 83.29万
• 依托单位: SEATTLE BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8204844
• 关键词: Academia; Affinity Chromatography; African; African Trypanosomiasis; Allergic Disease; Applied Research; Basic Science; Biochemistry; Biological Assay; Biology; Blood; Blood Circulation; Categories; Cellular Morphology; Characteristics; Clinical Trials; Cloning; Communicable Diseases; Communities; Computer Simulation; Data; Databases; Development; Discipline; Disclosure; Disease; Drug Delivery Systems; Drug Kinetics; Drug resistance; Enzymes; Escherichia coli; Essential Genes; Evaluation; Flow Cytometry; Gene Expression; Genes; Genome; Genomics; Germ; Goals; Growth; Human; Immune System Diseases; In Vitro; Industry; Infection; Informatics; International; Knowledge; Lead; Leishmania; Libraries; Metabolism; Microscopy; Molecular and Cellular Biology; Mus; National Institute of Allergy and Infectious Disease; Output; Parasites; Pharmaceutical Chemistry; Pharmaceutical Preparations; Plasmids; Preparation; Prevention program; Production; Proteins; RNA; RNA Interference; RNA analysis; Reagent; Recombinant Proteins; Research; Resources; Screening procedure; Set protein; Surveys; System; Technology; Testing; Time; Toxic effect; Toxicology; Transfection; Translating; Trypanosoma; Trypanosoma brucei brucei; Trypanosoma cruzi; Update; Validation; Wheat; assay development; base; cell growth; cost; drug candidate; drug development; drug discovery; genetic manipulation; genome sequencing; genome-wide; high throughput screening; in vivo; interest; knock-down; knowledge base; mouse model; novel; pathogen; programs; protein B; protein expression; protein structure; public health relevance; stable cell line; treatment program; vector; web site

DESCRIPTION (provided by applicant): Trypanosoma brucei, a WHO category one pathogen (emerging and uncontrolled), causes human African trypanosomiasis (HAT, aka African Sleeping Sickness), which is invariably fatal unless treated. New drugs are badly needed since current therapy is limited by toxicity, difficulty of administration, and spreading drug resistance. A wealth of knowledge has accumulated over the last 20-30 years on the molecular and cellular biology of T. brucei through a variety of hypothesis driven research programs. While several validated drug targets have arisen as a spin-off of these studies, there has been no systematic comprehensive effort to exploit the output of the genome projects and the recent advances in genetic manipulation of T. brucei to specifically identify and assess potential of drug targets. In addition, significant resources have been invested in setting up drug discovery programs including the building of facilities for high throughput screening (HTS) in several academic centers around the world. However, despite these efforts the list of validated drug targets that are both essential and likely to be "druggable" is small, and thus there are not enough high value targets that justify the considerable effort and resource of a HTS, or a dedicated drug discovery effort. This proposal is part of a coordinated effort by our US-based group (Stuart and Phillips) and our collaborators in the UK (Drs. M. Fergusson and J. Mottram), to undertake RNAi (interfering RNA) analysis in a genome-wide survey for essential, and "druggable" targets in T. brucei. A genome-wide data-base that lists key data required for target assessment in drug discovery is being compiled by the WHO/TDR and will be a key resource for target selection. While this proposal focuses on HAT, genetically validated targets in T. brucei may translate to T. cruzi and Leishmania species, and thus these studies will provide a resource for the entire kinetoplastid community. All data generated from the project will be provided to the tdrtargets.org curators for public distribution on that web site. The goals of this three year study are to 1) Select 200 - 300 genes in the T. brucei genome for RNAi essentiality analysis based on an assessment of potential "druggability"; 2) to determine if the selected genes are essential for the growth of blood form T. brucei parasites in vitro and in vivo by RNAi analysis; 3) to develop high-yield heterologous protein expression systems and suitable enzyme assays for genes that are shown to be essential; and 4) to organize data and reagents to provide distribution to the T. brucei research community. PUBLIC HEALTH RELEVANCE: The goal of the National Institute of Allergy and Infectious Diseases (NIAID) is to support basic and applied research leading to a better understanding of infectious, immunologic, and allergic diseases with the goal of generating new treatments or prevention programs for these diseases. The direct goal of this proposal is to identify new drug targets for the development of novel treatment programs against several priority trypanosomal pathogens that have been of long standing interest in the NIAID portfolio.

描述（由申请人提供）：Brucei锥虫（Trypanosoma brucei）是一类一级病原体（出现和不受控制的），会导致非洲人类非洲锥虫病（帽子，又名非洲睡眠疾病），除非治疗，否则是致命的。由于当前的治疗受到毒性，施用难度和耐药性的限制，因此非常需要新药。在过去的20 - 30年中，大量知识通过各种假设驱动的研究计划积累了有关T. Brucei的分子和细胞生物学的知识。尽管已经作为这些研究的衍生出来了几个经过验证的药物靶标，但没有系统的全面努力来利用基因组项目的产出以及t. brucei的遗传操作的最新进展，以专门识别和评估药物的潜力。目标。此外，已经投入了大量资源来制定药物发现计划，包括在全球几个学术中心建立高通量筛查设施（HTS）。但是，尽管进行了这些努力，但既重要又可能是“可药物”的经过验证的药物靶标的清单很小，因此没有足够的高价值目标证明HTS的巨大努力和资源是合理的，或者是专门的药物发现努力。该提案是我们的美国基于美国集团（Stuart和Phillips）以及我们在英国的合作者（M. Fergusson博士和J. Mottram）进行协调的一部分，以在全基因组中进行RNAi（干扰RNA）分析T. Brucei的必需和“可药物”目标的调查。 WHO/TDR汇总了列出药物发现目标评估所需的关键数据的全基因组数据库，将成为目标选择的关键资源。尽管该提案的重点是HAT，但T. Brucei的遗传验证靶标可能会转化为T. Cruzi和Leishmania物种，因此这些研究将为整个Kinetoplastid社区提供资源。从项目生成的所有数据都将提供给tdrtargets.org策展人在该网站上进行公开分发。这三年研究的目标是1）基于对潜在的“可药物性”的评估，在T. brucei基因组中选择200-300个基因进行RNAi的必要性分析； 2）确定所选基因在体外和体内通过RNAi分析在体外和体内生长至关重要； 3）开发高收益异源蛋白表达系统和适当的酶测定，以证明这是必不可少的基因； 4）组织数据和试剂，为T. Brucei研究社区提供分发。 公共卫生相关性：美国国家过敏和传染病研究所（NIAID）的目标是支持基础和应用研究，从而使人们对传染性，免疫学和过敏性疾病有更好的了解，目的是为这些创造新的治疗方法或预防计划疾病。该提案的直接目的是确定针对几种优先型锥虫病原体制定新的治疗计划的新药物，这些锥虫病原体对NIAID投资组合一直具有长期感兴趣。

项目成果

GMP synthase is essential for viability and infectivity of Trypanosoma brucei despite a redundant purine salvage pathway.

• DOI: 10.1111/mmi.13083
• 发表时间: 2015-09
• 期刊: Molecular microbiology
• 影响因子: 3.6
• 作者: Li Q;Leija C;Rijo-Ferreira F;Chen J;Cestari I;Stuart K;Tu BP;Phillips MA
• 通讯作者: Phillips MA

An Antiparasitic Compound from the Medicines for Malaria Venture Pathogen Box Promotes Leishmania Tubulin Polymerization.

• DOI: 10.1021/acsinfecdis.0c00122
• 发表时间: 2020-08-14
• 期刊: ACS infectious diseases
• 影响因子: 5.3
• 作者: Ullah I;Gahalawat S;Booshehri LM;Niederstrasser H;Majumdar S;Leija C;Bradford JM;Hu B;Ready JM;Wetzel DM
• 通讯作者: Wetzel DM