Animal Model Core E

动物模型核心E

• 批准号: 10438916
• 负责人: Clare Margaret Smith
• 金额: $ 17.08万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10438916
• 关键词: Address; Aerosols; Animal Model; Animals; Bacterial Genes; Bar Codes; CRISPR interference; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Data; Diagnostic; Diagnostic tests; Disease; Disease Outcome; Disease Progression; Dose; Encapsulated; Genes; Genetic; Genetic Variation; Goals; Growth; Human; Inbred Mouse; Inbred Strain; Individual; Infection; Inflammation; Integration Host Factors; Laboratories; Libraries; Link; Lipids; Lung; Lung infections; Mammalian Genetics; Measures; Metabolic; Modeling; Mouse Strains; Mus; Mutation; Mycobacterium tuberculosis; Natural regeneration; Outcome; Pathogenesis; Pathology; Patients; Phenotype; Process; Recombinants; Reproducibility; Resistance; Resources; Role; Serum; System; Testing; Tuberculosis; Universities; Variant; Virulence; animal model development; bacterial genetics; base; cohort; experimental study; genetic resource; genome-wide; human disease; in vivo; in vivo evaluation; interest; knock-down; lipidomics; mouse model; mutant; mycobacterial; pathogen; pressure; programs; response; stem; trait; transposon sequencing

Core E. Animal Model Core Project Leader: Clare Smith ABSTRACT Understanding the mechanisms through which Mycobacterium tuberculosis (Mtb) metabolites impact human tuberculosis (TB) disease requires implementation of model animal systems that are both tractable and faithfully replicate the TB disease states observed in humans. Here we propose to leverage the Collaborative Cross (CC), a genetically diverse panel of recombinant inbred mice that can be reproducibly and indefinitely regenerated. We have previously shown that the CC panel encompasses a broad spectrum of TB disease traits and infection microenvironments. CC mice provide a tractable model in which to study specific Mtb genetic-metabolite pairs, compared to standard inbred mice that show limited phenotypic variability. For example, in a genome-wide TnSeq experiment, we found that among 19 high value Mtb metabolic genes, only one controlled growth in the conventional C57BL/6J (BL6) mouse strain. However, more than half of mutants studied showed in vivo growth phenotypes when screened across CC mouse strains. Further, through study of the host genetic backgrounds in which individual bacterial genes do or do not control in vivo Mtb survival, the Smith laboratory can begin to study host factors in control of Mtb response. The Animal Core E will conduct experimental infection approaches to support Projects 1 and 3 that focus on virulence associated lipids and diagnostics, respectively. To support identification of Mtb metabolites as diagnostic tests in Project 3, we will characterize lungs and serum from mice infected with a high and low burden of Mtb. Supporting efforts to understand the role of host pressure on the mycobacterial envelope content in Project 1, we will produce Mtb strains passaged in vivo in mice. Extending our existing Tnseq approach to identify mycobacterial genes that control Mtb growth in vivo, we will select and test up 50 pooled Mtb CRISPR knockdown strains for pulmonary infection in CC strains. From these, five Mtb CRISPR knockdown strains with the strongest in vivo growth phenotypes will be further studied in detail as single gene knockdowns to determine their specific functions.

核心E.动物模型核心 项目负责人：克莱尔·史密斯（Clare Smith） 抽象的 了解结核分枝杆菌（MTB）代谢产物的机制 结核病（结核病）疾病需要实施既有瓦特易变的模型动物系统 忠实地复制人类观察到的结核病疾病状态。在这里，我们建议利用协作 Cross（CC），一种遗传多样的重组近交小鼠，可以无限地可重复地 再生。我们以前已经表明，CC面板包括大量结核病疾病 特质和感染微环境。 CC小鼠提供了一个可探讨的模型，其中研究特定的MTB 与表现有限的表型变异性的标准近交小鼠相比，遗传代谢物对。为了 例如，在全基因组TNSEQ实验中，我们发现在19个高价值MTB代谢基因中，仅在 常规C57BL/6J（BL6）小鼠应变中的一种受控生长。但是，超过一半的突变体 研究表明，当跨CC小鼠菌株筛选时，体内生长表型。此外，通过研究 单个细菌基因在体内MTB存活中所做或不控制的宿主遗传背景， 史密斯实验室可以开始研究控制MTB反应的宿主因素。动物核心E将进行 支持项目1和3的实验感染方法，专注于毒力相关的脂质和 诊断分别。为了支持将MTB代谢物作为项目3中的诊断测试的鉴定，我们将 表征来自MTB负担高和低负担的小鼠的肺和血清。支持努力 了解宿主压力对项目1中的分枝杆菌包膜含量的作用，我们将产生MTB 小鼠体内的菌株通过体内传递。扩展我们现有的TNSEQ方法来识别分枝杆菌基因 控制MTB在体内的增长，我们将选择并测试50个合并的MTB CRISPR敲低菌株 CC菌株感染。从中，五个MTB CRISPR敲除菌株具有最强的体内增长 表型将作为单个基因敲低进一步研究，以确定其特定功能。