Mouse modeling of a human STING gene variant for infectious disease

人类 STING 基因变体感染性疾病的小鼠模型

• 批准号: 8519291
• 负责人: John C Cambier
• 金额: $ 21.79万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8519291
• 关键词: Affect; American; Amino Acid Sequence; Amino Acid Sequence Homology; Amino Acids; Animal Model; Bacterial Infections; C-terminal; Communicable Diseases; Cues; Cyclic GMP; Cytoplasmic Tail; Defect; Development; Disadvantaged; Disease; Disease susceptibility; Dominant-Negative Mutation; Endoplasmic Reticulum; Exons; Fibroblasts; Future; Generations; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Goals; Health; Healthcare; Host Defense; Human; Human Genetics; Human Genome; Immune response; Immunity; In Vitro; Individual; Infection; Interferons; Introns; Knock-in Mouse; Knowledge; Listeria monocytogenes; Mediating; Membrane; Methods; Microbe; Mitochondria; Modeling; Molecular; Mus; N-terminal; Names; Outer Mitochondrial Membrane; Positioning Attribute; Predisposition; Production; Proteins; Relative (related person); Reporting; Research; Residual state; Resistance to infection; Risk; Role; Second Messenger Systems; Signal Pathway; Signal Transduction; Site; Sting Injury; Structure; Testing; Time; Transgenic Mice; Transmembrane Domain; Variant; Viral; Virus Diseases; base; experience; gene function; genetic variant; genome wide association study; human disease; human study; in vivo; inhibitor/antagonist; killings; macrophage; man; microorganism; mouse genome; mouse model; novel; pathogen; positional cloning; predictive modeling; response; second messenger; tool

DESCRIPTION (provided by applicant): Infectious diseases kill more people than any other single cause. Genome-Wide-Association-Studies in humans have demonstrated that genetic variations in genes involved in host defense can affect individuals' susceptibility to infection. I most cases, such reverse genetic approaches to understanding genetic contributions to disease are dependent on animal models that allow analysis of the effects of genetic variance in the absence of other variables. The lack of such models and the knowledge they yield poses a significant barrier for the future development of personalized treatment and health care for individuals carrying these genetic variations. Here we propose the construction of mouse gene knockin models to study the function of two variants of the human gene that encodes STING, a type I IFN stimulator that in mice is essential for defense against viral infections. We recently reported that there are two STING variants in man, each encoding three amino-acid changes (R71H-G230A-R293Q). We refer to these as WT and HAQ. Our in vitro studies have demonstrated that relative to WT the HAQ-STING variant has lost >90% of the ability to stimulate type I IFN production upon pathogen infection. Furthermore, the HAQ-STING variant has a dominant negative functional effect in vitro. Surprisingly, ~20% of Americans carry at least one copy of the HAQ-sting variant. It is our long-term goal to understand the molecular mechanisms and in vivo function of this frequent, potentially nonfunctional, human STING variant that may affect disease susceptibility of millions of Americans. To achieve this goal, we propose to develop two mouse models for the human STING variants. The two models will include 1) a minimalist knockin of SNPs into the mouse gene and 2) a knockin of SNPs plus unique human contextual sequence of potential importance in function. It is noteworthy that mouse and human STING proteins share 83% amino acid sequence homology. Thus in the first model (mHAQ mouse), we will make a STING knock-in in which the mouse equivalent amino acids will be changed to those in human HAQ-STING (Aim 1). In the second model we will accommodate the limited regions of non-homology, yet take advantage of the fact that mouse and human STING genes have the same exon - intron structure. We will make a STING knock-in that expresses a chimeric form of human-mouse STING, in which the entire N-terminal 172aa is replaced by the N-terminal of 173aa of human STING (cHAQ mouse) (Aim 2). This mouse will also contain G230A and R293Q. Appropriate WT mice will also be constructed. As time permits we will use these models to study the effects of HAQ on resistance to infection and on STING signaling function.

描述（由申请人提供）：传染病比任何其他单一原因杀死更多的人。全基因组关联-人类研究表明，参与宿主防御的基因的遗传变异会影响个体对感染的易感性。在大多数情况下，这种理解遗传对疾病的贡献的反向遗传方法依赖于动物模型，该动物模型允许在没有其他变量的情况下分析遗传方差的影响。缺乏这样的模型和知识，他们产生的个人携带这些遗传变异的个性化治疗和保健的未来发展构成了重大障碍。在这里，我们提出了小鼠基因敲入模型的建设，以研究人类基因的两种变体的功能，编码STING，I型IFN刺激剂，在小鼠中是必不可少的防御病毒感染。我们最近报道了在人类中存在两种STING变体，每种变体编码三个氨基酸变化（R71 H-G230 A-R293 Q）。我们称之为WT和HAQ。我们的体外研究表明，相对于WT，HAQ-STING变体在病原体感染后刺激I型IFN产生的能力丧失了>90%。此外，HAQ-STING变体在体外具有显性负功能效应。令人惊讶的是，约20%的美国人携带至少一个HAQ-sting变体的拷贝。我们的长期目标是了解这种可能影响数百万美国人疾病易感性的常见的、潜在的非功能性人类STING变体的分子机制和体内功能。为了实现这一目标，我们建议为人类STING变体开发两种小鼠模型。这两个模型将包括1）SNP敲入小鼠基因的最低限度敲入和2）SNP敲入加上在功能上具有潜在重要性的独特人类背景序列。值得注意的是，小鼠和人STING蛋白共享83%的氨基酸序列同源性。因此，在第一个模型（mHAQ小鼠）中，我们将进行STING敲入，其中小鼠等同氨基酸将改变为人HAQ-STING中的那些（Aim 1）。在第二个模型中，我们将容纳有限的非同源区域，但利用小鼠和人STING基因具有相同外显子-内含子结构的事实。我们将制备表达嵌合形式的人-小鼠STING的STING敲入，其中整个N-末端172 aa被人STING（cHAQ小鼠）的173 aa的N-末端替换（目的2）。该小鼠还将含有G230 A和R293 Q。还将构建适当的WT小鼠。如果时间允许，我们将使用这些模型来研究HAQ对感染抗性和STING信号传导功能的影响。

项目成果

Apoptotic caspases suppress mtDNA-induced STING-mediated type I IFN production.

• DOI: 10.1016/j.cell.2014.11.036
• 发表时间: 2014-12-18
• 期刊: Cell
• 影响因子: 64.5
• 作者: White MJ;McArthur K;Metcalf D;Lane RM;Cambier JC;Herold MJ;van Delft MF;Bedoui S;Lessene G;Ritchie ME;Huang DC;Kile BT
• 通讯作者: Kile BT