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. !

项目概要： 这种竞争性更新的一个核心科学问题是对炎症的机制理解 对胞质 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 万年前与经典近交系 (C57BL6) 小鼠分化的 MOLF 品系的衍生小鼠。 为该应用提供初步数据的表型是 MOLF 中干扰素产生的缺陷 巨噬细胞对 DNA 病原体或胞质 DNA 做出反应 - 一种由亚等位基因引起的病症 斯汀的。尽管无法产生 IFN，激活的 MOLF 巨噬细胞仍会过量产生 IL-6。其他野生- 衍生菌株对 DNA 病毒和 DNA 表现出类似的反应偏差。根据这些数据，我们 假设为了避免 IFN 介导的过度炎症，DNA 传感途径必须具有 进化来补充 IFN 的产生，或许还可以使用危险性较低的替代品，例如 IL-6。据此，我们 将确定野生小鼠如何对具有低 IFN 但高 IL-6 的胞质 DNA 做出反应——一系列 调查最终将确定导致 IL-6 过量产生的基因（位点）。在第一个目标中，我们将 检验以下假设：STING 在 ER 中的保留是将 DNA 反应从 IFN 至 IL-6 的产生。其次，我们将研究其他 DNA 传感器和途径的潜在贡献 MOLF 中 STING 介导的 DNA 反应。最后，我们将使用STING同源（B6.StingMOLF/MOLF）小鼠， 它们对 DNA 完全没有反应，无法进行遗传图谱和识别赋予基因的基因 MOLF 中对 DNA 的反应导致 IL-6 过量产生。在性状遗传分析的扩展中，我们将研究 DNAse2-/- Sting MOLF/- 小鼠对 DNA 的反应。这些小鼠被从 DNAse2-/- -相关性中拯救出来 尽管胚胎中炎症细胞因子水平较高，但 Sting 的亚等位基因仍具有胚胎致死率 DNAse2-/- Sting MOLF/- 雌性的血液和胚胎致死率。 这些数据 建议一些 这些小鼠中存在未表征的 STING 介导的信号传导，这将在实验的目标 2 中进行研究 提议。首先，我们将鉴定 DNA 反应细胞，并使用单细胞 RNA 测序分析 这些细胞识别具有与炎症特征和 DNA 相关的表达水平的基因 回应。尽管仅限于 eQTL（表达数量性状位点），但单细胞关联 目标 2 中的研究将有可能揭示表型和基因之间的所有关联，其中一些关联 不一定会映射到目标 1 中，并且可能是完全新颖的。最后，基于胚胎致死率 DNAse2-/- Sting MOLF/- 雌性中，我们将研究 X 连锁 Tlr7 和 Tlr8 的潜在贡献 进入 STING 介导的反应。通过研究 MOLF 中 DNA 反应的机制，我们希望 可以更好地了解人类干扰素病患者中存在的病理多样性。 ！

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