Animal Model Core E

动物模型核心E

基本信息

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
项目类别：
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

项目摘要

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·史密斯抽象的了解结核分枝杆菌 (Mtb) 代谢物影响人类的机制结核病 (TB) 需要实施既易于处理又易于控制的模型动物系统忠实地复制了在人类中观察到的结核病状态。在这里，我们建议利用协作 Cross (CC)，一组基因多样化的重组近交小鼠，可重复且无限期地再生了。我们之前已经表明，CC 组合涵盖了广泛的结核病特征和感染微环境。 CC 小鼠提供了一个易于处理的模型来研究特定的 Mtb 与显示有限表型变异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 个高价值的 Mtb 代谢基因中，只有一种控制传统 C57BL/6J (BL6) 小鼠品系的生长。然而，超过一半的突变体研究显示在对 CC 小鼠品系进行筛选时显示了体内生长表型。进一步地，通过研究宿主遗传背景，其中单个细菌基因控制或不控制体内 Mtb 存活，史密斯实验室可以开始研究控制结核分枝杆菌反应的宿主因素。 Animal Core E 将进行支持项目 1 和 3 的实验感染方法，重点关注与毒力相关的脂质和分别进行诊断。为了支持 Mtb 代谢物的鉴定作为项目 3 中的诊断测试，我们将表征感染高负荷和低负荷 Mtb 的小鼠的肺部和血清。支持努力了解宿主压力对项目1中分枝杆菌包膜含量的作用，我们将生产Mtb 菌株在小鼠体内传代。扩展我们现有的 Tseq 方法来识别分枝杆菌基因为了控制体内 Mtb 的生长，我们将选择并测试 50 个合并的 Mtb CRISPR 敲低菌株，用于肺部治疗 CC 菌株感染。其中，五种 Mtb CRISPR 敲除菌株具有最强的体内生长能力表型将作为单基因敲低进一步详细研究，以确定其特定功能。