Genetic Analysis of Inflammatory Responses in Wild Derived Mice

野生小鼠炎症反应的遗传分析

• 批准号: 10534173
• 负责人: Alexander Poltorak
• 金额: $ 76.97万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10534173
• 关键词: Affect; Alleles; Autoimmune; Biological Response Modifiers; Blood; Candidate Disease Gene; Cells; Code; Compensation; Complement; Cytosol; DNA; DNA Repair; DNA Viruses; DNA analysis; Dangerousness; Data; Defect; Embryo; Endosomes; Enzymes; Etiology; Exhibits; Failure; Female; Genes; Genetic; Genetic Recombination; Goals; High Prevalence; Homeostasis; Human; IL6 gene; Immune response; In Vitro; Inbreeding; Individual; Inflammation; Inflammatory; Inflammatory Response; Inherited; Interferon Type I; Interferon alpha; Interferons; Interleukin-6; Invaded; Investigation; Knockout Mice; Lead; Link; Macrophage; Maps; Mediating; Mediator; Meiotic Recombination; Messenger RNA; Mus; Mutation; Normal Statistical Distribution; Nucleic Acids; Outcome; Pathogenicity; Pathology; Pathway interactions; Patients; Phenotype; Production; Progress Reports; Proteins; Quantitative Trait Loci; Regulation; Role; STING1 gene; Signal Transduction; Stimulator of Interferon Genes; Sting Injury; TREX1 gene; Testing; Toxic effect; congenic; cytokine; cytotoxicity; differential expression; gene cloning; genetic analysis; immunopathology; in vivo; insight; loss of function; novel; pathogen; positional cloning; response; sensor; single cell analysis; single-cell RNA sequencing; trait

PROJECT SUMMARY: A central scientific question of this competing renewal is the mechanistic understanding of the inflammatory responses to cytosolic DNA. To achieve our goals, we continue using the forward genetic analysis in the wild- derived mice of the MOLF strain that diverged from classical inbred (C57BL6) mice about 1 million years ago. A phenotype that provides preliminary data for this application is the defective IFN production in MOLF macrophages in response to DNA pathogens or cytosolic DNA - a condition conferred by a hypomorphic allele of Sting. In spite of the failure to produce IFN, activated MOLF macrophages overproduce IL-6. Other wild- derived strains exhibit similar skewing of responses to DNA-viruses and DNA. Based on these data, we hypothesized that in order to avoid IFN-mediated excessive inflammation, DNA sensing pathways must have evolved to complement IFN production, perhaps with less dangerous substitutes such as IL-6. Accordingly, we will determine how wild derived mice respond to cytosolic DNA with low IFN but high IL-6--a line of investigation that will ultimately identify genes (loci) responsible for IL-6 overproduction. In the first Aim, we will test the hypothesis that retention of STING in the ER is the mechanism of switching the DNA-responses from IFN to IL-6 production. Second, we will examine the potential contribution of other DNA-sensors and pathways into STING-mediated DNA-responses in MOLF. Finally, we will use STING congenic (B6.StingMOLF/MOLF) mice, which are completely non-responsive to DNA to genetically map and identify gene(s) that confer overproduction of IL-6 in MOLF in response to DNA. In extension of genetic analysis of the trait, we will study the responses to DNA in DNAse2-/- Sting MOLF/- mice. These mice are rescued from DNAse2-/- -associated embryonic lethality with the hypomorphic allele of Sting despite high levels of inflammatory cytokines in the blood and embryonic lethality of the DNAse2-/- Sting MOLF/- females. These data suggest that some uncharacterized STING-mediated signaling exists in these mice, which will be investigated in Aim 2 of the proposal. First, we will identify the DNA-responsive cells, and use single cell RNA-sequencing analysis in these cells to identify genes with expression levels associated with the inflammatory signature and DNA- responses. Despite being confined to eQTL (expression Quantitative Trait Loci), the single cell association studies in Aim 2 will potentially reveal all associations between the phenotype and the genes, some of which will not necessarily be mapped in Aim 1 and could be completely novel. Finally, based on embryonic lethality of the DNAse2-/- Sting MOLF/- females, we will investigate potential contributions of the X-linked Tlr7 and Tlr8 into STING-mediated responses. By investigating the mechanism of DNA responses in MOLF, we hope to provide better insight into the diversity of pathologies present in human patients with interferonopathies. !

项目总结： 这种相互竞争的更新的一个中心科学问题是对炎症性疾病的机械理解 对胞浆DNA的反应。为了实现我们的目标，我们继续在野外使用正向遗传分析- 大约100万年前从经典近亲交配(C57BL6)小鼠分化而来的Molf品系小鼠。 为这一应用提供初步数据的一个表型是Molf中有缺陷的干扰素生产 巨噬细胞对DNA病原体或胞浆DNA的反应--一种由亚型等位基因引起的状态 斯汀。尽管不能产生干扰素，但激活的Molf巨噬细胞会过度产生IL-6。其他狂野的- 衍生菌株对DNA病毒和DNA的反应表现出类似的偏斜。根据这些数据，我们 假设为了避免干扰素介导的过度炎症，DNA传感通路必须有 进化为补充干扰素的产生，可能使用不太危险的替代品，如IL-6。因此，我们 将确定野生来源的小鼠对含有低干扰素但高IL-6的胞浆DNA的反应 最终将确定导致IL-6过度生产的基因(基因座)的研究。在第一个目标中，我们将 检验假设，内质网中刺突的保留是将DNA反应从 干扰素对IL-6的产生。第二，我们将研究其他DNA传感器和通路的潜在贡献。 在Molf中转化为刺激性DNA反应。最后，我们将使用STING同源(B6.StingMOLF/Molf)小鼠， 对dna完全不敏感的基因进行遗传定位和鉴定(S) DNA诱导Molf细胞过度产生IL-6。在性状遗传分析的延伸中，我们将研究 DNAse2-/-Sting Molf/-小鼠对DNA的反应这些小鼠被从DNAse2-/--相关的DNA中拯救出来 与Sting亚型等位基因有关的胚胎致死性，尽管在 DNAse2-/-Sting Molf/-雌性的血液和胚胎致死性 这些数据 建议一些人 在这些小鼠中存在未知的刺痛介导的信号，这将在 求婚。首先，我们将鉴定DNA反应细胞，并使用单细胞RNA测序分析在 这些细胞识别与炎症信号和DNA相关的表达水平的基因- 回应。尽管局限于eQTL(表达数量性状基因座)，但单细胞关联 AIM 2的研究可能会揭示表型和基因之间的所有关联，其中一些 不一定会被映射到目标1中，并且可能是完全新颖的。最后，基于胚胎致命性 在DNAse2-/-Sting Molf/-雌体中，我们将研究X连锁的TLR7和TLR8的潜在贡献 以毒刺为媒介的反应。通过研究Molf中DNA反应的机制，我们希望能够 更好地洞察人类干扰素病患者存在的各种病理变化。好了！

项目成果

Cutting Edge: Activation of STING in T Cells Induces Type I IFN Responses and Cell Death.

最前沿：T 细胞中 STING 的激活可诱导 I 型 IFN 反应和细胞死亡。

• DOI: 10.4049/jimmunol.1601999
• 发表时间: 2017-07-15
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Larkin B;Ilyukha V;Sorokin M;Buzdin A;Vannier E;Poltorak A
• 通讯作者: Poltorak A

IRAK-2 regulates IL-1-mediated pathogenic Th17 cell development in helminthic infection.

• DOI: 10.1371/journal.ppat.1002272
• 发表时间: 2011-10
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Smith PM;Jacque B;Conner JR;Poltorak A;Stadecker MJ
• 通讯作者: Stadecker MJ

ZBP1 promotes inflammatory responses downstream of TLR3/TLR4 via timely delivery of RIPK1 to TRIF.

• DOI: 10.1073/pnas.2113872119
• 发表时间: 2022-06-14
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1

The PYRIN connection: novel players in innate immunity and inflammation.

pyrin的联系：先天免疫和炎症的新手。

• DOI: 10.1084/jem.20032234
• 发表时间: 2004-09-06
• 期刊: JOURNAL OF EXPERIMENTAL MEDICINE
• 影响因子: 15.3
• 作者: Stehlik, C;Reed, JC
• 通讯作者: Reed, JC

Endothelial STING controls T cell transmigration in an IFNI-dependent manner.

• DOI: 10.1172/jci.insight.149346
• 发表时间: 2021-08-09
• 期刊: JCI insight
• 影响因子: 8
• 作者: Anastasiou M;Newton GA;Kaur K;Carrillo-Salinas FJ;Smolgovsky SA;Bayer AL;Ilyukha V;Sharma S;Poltorak A;Luscinskas FW;Alcaide P
• 通讯作者: Alcaide P